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Anti-proliferative and antioxidant activities of Saposhnikovia divaricata
Abstract
The dry root of Saposhnikovia divaricata (Turcz.) Schischk. (SD, syn. Ledebouriella divaricata (Turcz.); Umbelliferae), Siler, a perennial herb of the carrot family, is also known as Fang Feng in traditional Chinese herbal medicine. It is a herbal ingredient included in many polyherb formulae. This study investigated the in vitro anti-proliferative, antioxidant and anti-inflammatory activities of the SD extract (1 g/10 ml 70% ethanol). IC50 (50% inhibition) is estimated at 1/300, 1/1400, 1/250 and 1/600 dilutions, for the K562, HL60, MCF7 and MDA-MB-468 cell lines, respectively. The combination of non-cytotoxic concentrations of SD with chemotherapeutic drugs such as camptothecin or paclitaxel showed additive anti-proliferative effects on K562, HL60 and MCF7 cells, and antagonistic effects on MDA-MB-468 cells. At a dilution of 1/2000, SD induced a differentiation of 17.5+/-2.5% in HL60 cells along the granulocyte lineage compared to 2.8+/-0.8% in the untreated controls, but not along the monocyte/macrophage lineage. At non-cytotoxic 1/10000, 1/5000 and 1/2000 dilutions, the SD extract did not affect nitric oxide (NO) production by non-stimulated RAW 264.7 cells, but dose-dependently and significantly reduced NO production by lipopolysaccharide (LPS)-activated RAW 264.7 cells. RT-PCR analyses showed that SD at a dilution of 1/2000 did not affect TNFalpha, IL-1 beta, iNOS and COX-2 mRNA expression in RAW 264.7 cells compared to the unstimulated controls, but significantly reduced (p<0.05) iNOS and its mRNA expression in LPS-activated cells. It is concluded that the SD ethanol extract possesses strong anti-proliferative properties against several human tumor cell lines, a mild granulocyte differentiation inducing property on HL60 cells, and potent antioxidant, anti-inflammatory and protective properties on LPS-activated RAW 264.7 cells. Further research is required in order to identify the major ingredients present in the Saposhnikovia divaricata root and rhizome showing the observed activities.
... It has been widely applied for the treatment of pyrexia, rheumatism, headache, vertigo, body aches, and arthralgia as an ingredient of several formulae in traditional medicine [1]. Previous pharmacological studies have indicated that SR possesses various pharmacological effects, including analgesic, anti-oxidant, anti-proliferative, anti-allergic, anti-inflammatory, and anti-tumor activities [2][3][4][5][6][7]. Active constituents in SR include chromones, coumarins, polyacetylenes, polysaccharides, and essential oils, among which major chromones such as prim-O-glucosylcimifugin (1) and 4′-O-β-d-glucosyl-5-O-methylvisamminol (3) are most relevant to anti-inflammatory, analgesic, and anti-allergic effects [7][8][9][10]. ...
... A portion (109 g) of the MeOH extract was subjected to DIAION HP-21 column (500 g) and eluted with a gradient mixture of H 2 O/MeOH (100:0 to 0:100, v/v) to obtain five subfractions (Frs. [1][2][3][4][5]. Fr. 5 (100% MeOH eluent, 8.0 g) was dissolved Table 1 List of the plant specimens collected from Mongolia in 2015 and 2019 and the Saposhnikoviae Radix samples collected from China a Specimens M1-M43, C1-C2 and SR samples C3-C8 used in this study were listed in our previous report [16] b Latitude, longitude, and altitude were not clear (-) c Specimens with flowers (F) and without flowers (NF); SR samples derived from wild plant (W) and cultivated plant (C) d Diameter was measured at the thickest position of the root. ...
... The data matrix of peak areas of the compounds (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14) was normalized by the Z score transformation prior to the multivariate statistical analysis. PCA was performed on 42 plant specimens (except for 2 specimens; M65, M66) collected in the seven different regions of Mongolia. ...
Saposhnikoviae Radix (SR), derived from the dried root and rhizome of Saposhnikovia divaricata, is a popular crude drug used in traditional Chinese and Japanese medicine. To evaluate the metabolites of S. divaricata roots from Mongolia and to investigate their geographical variation, we developed the HPLC method, determined the contents of 9 chromones and 4 coumarins, and conducted multivariate statistical analysis. All Mongolian specimens contained prim-O-glucosylcimifugin (1) and 4′-O-β-d-glucosyl-5-O-methylvisamminol (3), and their total amount (5.04–25.06 mg/g) exceeded the criterion assigned in the Chinese Pharmacopoeia. Moreover, the content of 1 (3.98–20.79 mg/g) was significantly higher in the Mongolian specimens than in Chinese SR samples. The specimens from Norovlin showed the highest contents of 1 and 3. The total levels of dihydropyranochromones were higher in the specimens from Bayan-Uul. The orthogonal partial least squares-discriminant analysis revealed that the Mongolian specimens tended to be separated into three groups based on growing regions, in which several chromones contributed to each distribution. Furthermore, 1H NMR analysis revealed that Mongolian specimens had less amount of sucrose and a substantial amount of polyacetylenes. Thus, in this study, the chemical characteristics of Mongolian S. divaricata specimens were clarified and it was found that the specimens from the northeast part of Mongolia, including Norovlin, had the superior properties due to higher amounts of major chromones.
... Phytochemical studies have revealed that there are approximately 100 compounds with different structural patterns isolated from S. divaricata, including chromones, coumarins, acid esters, polyacetylenes, volatile oils, polysaccharides, and inorganic elements (1,4,(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28) as shown in Appendixes 3-9. ...
... (15) Despite their simple structure, they have good anti-inflammatory, free radical-scavenging, and immunostimulatory effects. (16)(17)(18) Among them, compounds 3, 7-10 are the primary chromones, which are abundant in S. divaricata, in particular, compounds 7 and 8. (19,20) Coumarins Coumarins, a type of benzopyrones containing a benzene ring fused to a pyrone ring, are the most abundant chemical constituents of S. divaricata. They have extremely diverse structures, which are divided into simple coumarins, pyranocoumarins, furanocoumarins, and other coumarins. ...
... (1,21,22) At present, a total of 35 coumarins, primarily furanocoumarins and pyranocoumarins, have been identified from S. divaricata (Appendixes 4 and 5). Among them, the 17 furanocoumarins isolated include bergapten (18), byakangelicin (19), deltoin (20), imperatorin (21), isoimperatorin (22), isobergapten (23), marmesin (24), methoxy-8-(3-hydroxymethylbut-2-enyloxy)-psoralen (25), nodakenetin (26), oxypeucedanin hydrate (27), phellopterin (28), psoralen (29), sapodivarin (30), xanthotoxin (31), 5-hydroxy-8-methoxypsoralen (32), nodakenin (33), and xanthoarnol (34). (23) Five simple coumarins isolated from the herb, including fraxidin (35), isofraxidin (36), scopoletin (37), umbelliferone (38) and 5-methoxy-7-(3, 3-dimethylallyloxy)-coumarin (39), have simple structures but show a variety of biological activities such as dispelling phlegm and antitumor activity. ...
Saposhnikovia divaricata (Turcz.) Schischk., a perennial herb belonging to the family Umbelliferae, is widely distributed in Northeast Asia. Its dried root (Radix Saposhnikoviae) is used as a Chinese herbal medicine for the treatment of immune system, nervous system, and respiratory diseases. Phytochemical and pharmacological studies have shown that the main constituents of S. divaricata are chromones, coumarins, acid esters, and polyacetylenes, and these compounds exhibited significant anti-inflammatory, analgesic, antioxidant, antiproliferative, antitumor, and immunoregulatory activities. The purpose of this review is to provide comprehensive information on the botanical characterization and distribution, traditional use and ethnopharmacology, phytochemistry, and pharmacology of S. divaricata for further study concerning its mechanism of action and development of better therapeutic agents and health products from S. divaricata.
... | www.phcog.net Figure 2: Effects of prim-O-glucosylcimifugin on cell viability with a Cell Counting Kit. RAW 264.7 cells were exposed with lipopolyssacharide (1 µg/mL) and various concentrations of prim-O-glucosylcimifugin (15,50 and 100 µg/mL) or dimethyl sulfoxide alone. Cell viability was assessed 24 h after treatment and expressed as percentage of the dimethyl sulfoxide control. ...
... Schischek (Umbelliferae), which is mainly distributed in Northeastern and Northern China. [9] RS is a well-known traditional Chinese medicine that has been commonly used in the clinical compound prescription to treat common colds, headache, (15,50, and 100 µg/mL). Cells treated with dimethyl sulfoxide were set as control. ...
... &&& P < 0.001 versus control; *P < 0.05, **P < 0.01, and ***P < 0.001 versus lipopolysaccharide-treated cells; # P < 0.05, ## P < 0.01, and ### P < 0.001 versus lipopolysaccharide and 15 µg/mL prim-O-glucosylcimifugin-treated cells; $$ P < 0.01 and $$$ P < 0.001 versus lipopolysaccharide and 50 µg/mL prim-O-glucosylcimifugin-treated cells rheumatoid arthritis, and eczema in China and other Asian countries. [9][10][11] Pharmacological experiments have demonstrated the anti-inflammatory, analgesic, antipyretic, antioxidation properties of RS. [10][11][12][13][14][15][16][17] Several kind of compounds have been isolated from RS, including chromones, coumarins, mannitol, glycoside, and polyacetylenes. [18,19] The major active constituents of RS are considered to be chromones and their glycosides. ...
Background
Radix Saposhnikoviae (RS) exerts anti-inflammatory, analgesic, antipyretic, antioxidation effects and has been used in traditional Chinese medicine to treat common colds, headache, and rheumatoid arthritis. Prim-O-glucosylcimifugin (POG) is the highest content chromone and one of the major active constituents in RS.
Objective
The study was aimed to explore the anti-inflammation effects of POG in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages.
Materials and Methods
Cell viability was detected by Cell Counting Kit-8 assay. Production of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 was assessed by enzyme-linked immunosorbent assay. Real-time polymerase chain reaction and Western blot were performed to analyze mRNA and protein levels, respectively.
Results
During the whole experiment, 15, 50, and 100 μg/mL of POG had no cytotoxicity on RAW 264.7 cells. POG dose-dependently inhibited the production of NO, TNF-α, IL-1β, and IL-6 that were induced by LPS. POG treatment downregulated the mRNA and protein expression inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2) in LPS-activated RAW 264.7 macrophages in a concentration-dependent manner. Furthermore, LPS-induced JAK2/STAT3 activation was prevented in RAW 264.7 macrophages by POG treatment. STAT3 overexpression significantly reversed the effects of POG on LPS-activated RAW 264.7 macrophages.
Conclusion
These results demonstrate that POG exerts anti-inflammatory effects through the inhibition of iNOS and COX-2 expression by inhibiting the phosphorylation of JAK2/STAT3.
SUMMARY
POG exerts anti-inflammatory effects in RAW 264.7 macrophages through the inhibition of iNOS and COX-2 expression by inhibiting JAK2/STAT3 signaling.
Abbreviations used: LPS: Lipopolyssacharide; NO: Nitric oxide; TNF-α: Tumor necrosis factor-α; IL: Interleukin; RS: Radix Saposhnikoviae; POG: Prim-O-glucosylcimifugin; iNOS: Inducible NO synthase; COX2: Cyclooxygenase; FBS: Fetal bovine serum; DMSO: Dimethylsulfoxide; CCK-8: Cell Counting Kit; RIPA: Radio immunoprecipitation assay buffer; ECL: Enhanced chemiluminescence; SD: Standard deviation; ELISA: Enzyme-Linked immunosorbent assay.
... Among 9 phytochemistry papers with in vitro studies [8][9][10][11][12][13][14][15][16] , 5 [11,[13][14][15][16] were selected based on their relevance, but the phytochemistry component did not adhere to the selection criteria. A total of 10 in vitro studies only were included in this review [17][18][19][20][21][22][23][24][25][26] . Fig. 1 illustrates the study selection process. ...
... A total of 9 chromones are identified in the included studies. These chromones are discovered to exhibit strong pharmacological activities in attenuating inflammatory [4] , scavenging free radical [20] , and inhibiting pain [8] in in vitro studies. ...
... Four cancer cell lines, i.e., human mammary adenocarcinoma MCF7 and MDA-MB-468, leukemia K562, and myelomonocytic HL60 cells, and murine RAW 264.7 macrophage/monocyte cells were tested in vitro with camptothecin (CAM), paclitaxel (PTX), and in combination with SD [20] . ...
Saposhnikoviae divaricata (Turcz.) Schischk (SD) is a traditional Chinese herb commonly used to treat clinical conditions such as rheumatism and allergic rhinitis. This review article evaluates a collection of works on in vitro and biochemical studies of SD. The discourse on the diverse class of chromones and coumarins in SD offers an insight to the pharmacological effects of these bioactive constituents as anti-inflammatory, analgesic, immunoregulatory, antioxidative, and anti-proliferative agents. It is highlighted that there is a structural relationship between the constituents and bioactive activities, which in effect provides a valid reasoning and reaffirm the use of SD in the treatment of the pathologies in Chinese medicine.
... The values of these indices were lower for 75% hydroethanolic extract and water extract and the lowest for juice. ABTS· + scavenging activity by 70% hydroethanolic extract from the root of S. divaricata was demonstrated earlier by other authors [18]. Our results indicate the highest content of hydrophilic and hydrophobic antioxidants in 50% hydroethanolic extract and the largest potential of this extract in decreasing oxidative stress in the organism. ...
... The results of the study of Tai and Cheung [18] indicate the potential applicability of 70% hydroethanolic extract from the root of S. divaricata as an element of combination therapy. This extract showed antiproliferative activities on HL60 human acute promyelocytic leukemia, MDA-MB-468 human mammary adenocarcinoma, K562 human chronic myelogenous leukemia and MCF7 human mammary adenocarcinoma cell lines. ...
Colorectal cancer ranks 3rd in terms of cancer incidence. Growth and development of colon cancer cells may be affected by juice and extracts from Saposhnikovia divaricata root. The objective of the research was to analyze the effect of S. divaricata juice and extracts on the viability, membrane integrity and types of cell death of Caco-2 cells. Juice and extracts were analyzed using Ultra-High Performance Liquid Chromatography-Mass Spectrometry (UHPLC-MS) and in respect of the presence of antioxidants, total carbohydrates, protein, fat and polyphenols. The contents of cimifugin β-D-glucopyranoside, cimifugin, 4-O-glucopyranosyl-5-O-methylvisamminol, imperatorin and protein were the highest in juice. 50% Hydroethanolic extract had the greatest antioxidant potential, concentration of polyphenols and fat. Water extract was characterized by the highest content of glutathione. Juice and 75% hydroethanolic extract contained the most carbohydrates. After the application of juice, 50% extract and the juice fraction containing the molecules with molecular weights >50 kDa, a decrease of the cell viability was noted. Juice and this extract exhibited the protective properties in relation to the cell membranes and they induced apoptosis. The knowledge of further mechanisms of anticancer activity of the examined products will allow to consider their use as part of combination therapy.
... [55][56][57] It has been demonstrated that the ethanol extract of Fang Feng possesses antioxidative and anti-inflammatory effects. 58 Later studies demonstrated that polyacetylene, imperatoin, and deltoin are potent iNOS inhibitors, they inhibit NO production which plays a major role in the early inflammatory cascade triggered by oxidative tissue injury. 56,59 ...
Equine recurrent airway obstruction and summer pasture-associated obstructive pulmonary disease are the major causes of chronic poor performance and exercise intolerance in athletic horses. Conventional treatment includes administration of a bronchodilator, an anti-inflammatory agent, and elimination of the inciting environment. Affected horses usually require life-long medication. If a corticosteroid is being used as the anti-inflammatory agent, an iatrogenic immune suppression and repression of the endogenous adrenocorticosteroid production are the most common side effects. Chinese veterinary herbal medicine provides an alternative long-term therapy with minimal side effects. Four common Chinese herbal formulas, Schisandra 5, Lily Combination, Bu Fei San, and Breath Easer have been shown to be useful for treating chronic respiratory disease. The pharmacological mechanisms of the phytochemical compounds present in 23 herbs found in these formulas are reviewed in this article. Active phytochemical compounds in the herbs are found in the form of primary and secondary metabolites. They may possess antimicrobial, antioxidant, anti-inflammatory, antihistamine, or antitussive properties alone or in combination. The evidence for a relevant pharmacological mechanism of the active phytochemical compounds in each herb validates the use of Chinese herbal supplements for treating chronic respiratory problems. These findings emphasize that the development of herbal formulas by ancient Chinese herbalists was conducted cautiously and based on a unique knowledge of pharmacology and toxicology.
... Flavonoid is polyphenol compound with many valuable biological functions, including antitumor, anti-oxidation, anti-inflammation and so on Liu et al., 2021;Zhang S. et al., 2021). Modern pharmacological studies show that chromogen ketone is the most important chemical component of S. divaricata, and chromogen ketones are mainly synthesized by flavonoid biosynthesis pathway (Tai and Cheung, 2007;Kreiner et al., 2017). It has been shown that phenylalanine and tyrosine are precursors of phenylpropane biosynthesis, which is then reacted with L-phenylalanin ammonialyase(PAL) to produce cinnamic acid and p-coumaric acid. ...
Introduction
Saposhnikovia divaricata is a traditional Chinese medicine in China, which is widely used in clinic. The root of S. divaricata is often used as medicine, but little research has been done on its other tissues.
Methods
In this study, the contents of root and leaf of S. divaricata were determined by HPLC, the differentially expressed genes were screened by transcriptome sequencing at molecular level, and then verified by network pharmacology.
Results
The results showed that the content of 4’-O-β-D-glucosyl-5-O-methylvisamminol in the leaves was significantly higher than that in the roots, which was about 3 times higher than that in the roots. In addition, 10 differentially expressed key enzyme genes were screened in plant hormone signal transduction, phenylpropanoid and flavonoid biosynthetic pathways. C4H and CYP98A were up-regulated in root, while F3H was down-regulated in root. They can be used as important candidate genes for the mechanism of quality difference of S. divaricata. Finally, network pharmacological validation showed that 5-O-methylvesamitol plays an important role in the treatment of ulcerative colitis.
Discussion
These findings not only provide insight into flavonoid biosynthesis in S. divaricata associated molecular regulation, but also provide a theoretical basis for the development and utilization of S. divaricata .
... Schischk, and is widely used in the clinical treatment of cold, headache, and skin pruritus [1]. Saposhnikoviae Radix has a variety of pharmacological activities, such as anti-inflammatory, bacteriostatic, anti-allergy, and anti-oxidation [2][3][4][5]. Saposhnikoviae Radix contains a variety of active ingredients, in which coumarins and chromogens have been studied extensively and work has focused on the structure and related activity of these compounds. As one of the main active components, Saposhnikoviae Radix polysaccharide (SP) has demonstrated a range of biological activities [6][7][8][9]. ...
PMP-HPLC, FT-IR, and HPSEC fingerprints of 10 batches of polysaccharides from Saposhnikoviae Radix with different production areas and harvest times have been prepared, and the chemometrics analysis was performed. The anti-allergic activity of 10 batches of Saposhnikoviae Radix polysaccharide (SP) was evaluated, and the spectrum-effect relationship of the 10 batches of SP was analyzed by gray correlation degree with the chromatographic fingerprint as the independent variable. The results showed that the PMP-HPLC, HPSEC, and FT-IR fingerprints of 10 batches of SP had a high similarity. Two monosaccharides (rhamnose and galactose), the polysaccharide fragment Mn = 8.67 × 106~9.56 × 106 Da, and the FT-IR absorption peak of 892 cm−1 can be used as the quality control markers of SPs. All 10 batches of SP could significantly inhibit the release of β-HEX in RBL-231 cells, and the polysaccharides harvested from Inner Mongolia in the winter had the best anti-allergic activity. The spectrum-effect relationship model showed that the monosaccharide composition and molecular weight were related to the anti-allergic activity of the SPs. Multiple fingerprints combined with spectrum-effect relationship analysis can evaluate and control the quality of SPs from the aspects of overall quality and efficacy, which has more application value.
... The dried roots are called Fang-Feng in China, Bang-Poong in Korea, and Bofu in Japan, and have been extensively used for treating arthralgia, headaches, rheumatism, stroke, fever, and allergic rhinitis [1]. Recently, studies investigating the chemical constituents of S. divaricata revealed that the main active components were chromones, coumarins, and volatile oils [2][3][4], which exhibited anti-proliferative and anti-oxidant, anti-bacterial and anti-tumor, anti-convulsant, anticoagulant, anti-inflammatory, and anti-pyretic properties [4][5][6][7][8]. However, little information is known regarding genetic diversity and evolution. ...
Saposhnikovia divaricata, a well-known Chinese medicinal herb, is the sole species under the genus Saposhnikovia of the Apiaceae subfamily Apioideae Drude. However, information regarding its genetic diversity and evolution is still limited. In this study, the first complete chloroplast genome (cpDNA) of wild S. divaricata was generated using de novo sequencing technology. Similar to the characteristics of Ledebouriella seseloides, the 147,834 bp-long S. divaricata cpDNA contained a large single copy, a small single copy, and two inverted repeat regions. A total of 85 protein-coding, 8 ribosomal RNA, and 36 transfer RNA genes were identified. Compared with five other species, the non-coding regions in the S. divaricata cpDNA exhibited greater variation than the coding regions. Several repeat sequences were also discovered, namely, 33 forward, 14 reverse, 3 complement, and 49 microsatellite repeats. Furthermore, phylogenetic analysis using 47 cpDNA sequences of Apioideae members revealed that L. seseloides and S. divaricata clustered together with a 100% bootstrap value, thereby supporting the validity of renaming L. seseloides to S. divaricata at the genomic level. Notably, S. divaricata was most closely related to Libanotis buchtormensis, which contradicts previous reports. Therefore, these findings provide a valuable foundation for future studies on the genetic diversity and evolution of S. divaricata.
... Although there is no approved hepatolipotoxic drug in the clinic, evidence emerged that phytochemicals from medicinal herbs own a vigorous capacity to ameliorate lipotoxicity [17]. Among those plant-derived compounds, cimifugin (Cim) is a coumarin derivative extracted from the root of Saposhnikovia divaricata, which is a traditional Chinese medical herb called Fang-feng (防风) with the usage history for the clinical therapy of allergy, rheumatism, headache, and convulsion, especially for allergic dermatitis and skin pruritus [18]. It has been known that Cim possesses a strong ability to prevent inflammation, allergy, and oxidative stress [19][20][21][22]. ...
Objective:
Hepatic metabolic disorder induced by lipotoxicity plays a detrimental role in metabolic fatty liver disease pathogenesis. Cimifugin (Cim), a coumarin derivative extracted from the root of Saposhnikovia divaricata, possesses multiple biological properties against inflammation, allergy, and oxidative stress. However, limited study has addressed the hepatoprotective role of Cim. Here, we investigate the protective effect of Cim against lipotoxicity-induced cytotoxicity and steatosis in hepatocytes and clarify its potential mechanisms.
Methods:
AML-12, a nontransformed mouse hepatocyte cell line, was employed in this study. The cells were incubated with palmitate or oleate to imitate hepatotoxicity or steatosis model, respectively.
Results:
Cim significantly reversed palmitate-induced hepatocellular injury in a dose-dependent manner, accompanied by improvements in oxidative stress and mitochondrial damage. Cim pretreatment reversed palmitate-stimulated TLR4/p38 MAPK activation and SIRT1 reduction without affecting JNK, ERK1/2, and AMPK pathways. The hepatoprotective effects of Cim were abolished either through activating TLR4/p38 by their pharmacological agonists or genetical silencing SIRT1 via special siRNA, indicating a mechanistic involvement. Moreover, Cim treatment improved oleate-induced hepatocellular lipid accumulation, which could be blocked by either TLR4 stimulation or SIRT1 knockdown. We observed that SIRT1 was a potential target of TLR4 in palmitate-treated hepatocytes, since TLR4 agonist LPS aggravated, whereas TLR4 antagonist CLI-095 alleviated palmitate-decreased SIRT1 expression. SIRT1 knockdown did not affect palmitate-induced TLR4. In addition, TLR4 activation by LPS significantly abolished Cim-protected SIRT1 reduction induced by palmitate. These results collaboratively indicated that TLR4-regulated SIRT1 pathways was mechanistically involved in the protective effects of Cim against lipotoxicity.
Conclusion:
In brief, we demonstrate the protective effects of Cim against lipotoxicity-induced cell death and steatosis in hepatocytes. TLR4-regulated p38 MAPK and SIRT1 pathways are involved in Cim-protected hepatic lipotoxicity. Cim is a potential candidate for improving hepatic metabolic disorders mediated by lipotoxicity.
... В настоящее время в традиционной китайской медицине корни данного растения используются в виде отвара в качестве жаропонижающего средства, а этанольный экстракт обладает противовоспалительным и противопростудным действиями [1, с. 20]. Этанольный экстракт сапожниковии растопыренной корней обладает антиоксидантной активностью in vitro, значительно снижая продукцию NO клетками RAW, активированными LPS [21]. В научном исследовании M. Kim и др. ...
... A group of Chinese researchers claimed that 0.1-0.5% by weight of radix saposhnikovia (the root of the perennial herbaceous plant Saposhnikovia divaricata) has high anti-wart properties. Hence, they made and tried this on the wart as an application with podophyllotoxin (0.02-0.05%), berberine (0.02-0.05%), atractylodin (0.1-0.2%) and alkannin (0.1-0.5%) with good outcomes [50,51]. A patent for another traditional Chinese topical remedy for warts was applied for by a team of researchers in China. ...
Warts, small skin protuberances caused by human papillomavirus, are not life-threatening, but can be unsightly, embarrassing, uncomfortable and potentially contagious. They may regress naturally but over a prolonged time. Hence wart-afflicted people seek prescribed or over-the-counter therapies. Out of the innumerable marketable anti-wart medications, only a few are effective. Besides, frequent applications over months and certain undesirable side effects limit their success rates. Recurrences are also known. Hence the search for an effective, safe, cheap and quick medicine that is easy to apply continues. A number of patents for anti-wart therapies have been filed in the last few years. In this article, we enlist the recent anti-wart patents and find out those holding promise in the future.
... In recent years, the pharmacological activity and chemical constituents of RS have been extensively studied by researchers worldwide (Kreiner et al., 2017). Anti-inflammatory, analgesic, antipyretic, antiallergic, antioxidant, antitumoral and anti-viral activities have been shown on different Saposhnikovia extracts in previous studies (Wang et al., , 1999Okuyama et al., 2001;Kuo et al., 2002;Tal and Cheung, 2007;Khan et al., 2013;Kong et al., 2013;Salman and Shik, 2013;Yua et al., 2015;Chun et al., 2016;Yang et al., 2015). The study of the chemical composition of Saposhnikovia divaricata led to the release of chromones, coumarins, unsaturated fatty acids, volatile oils, and polyacetylene compounds (Sasaki et al., 1982;Baba et al., 1987;Okuyama et al., 2001;Kang et al., 2008;Yu et al., 2009;Gui et al., 2011;Kim et al., 2011;Yang et al., 2015;Chen et al., 2018). ...
... В недавних исследованиях Kim M. и др. по взаимосвязи между антимикробной и антиоксидантной активностью и общим содержанием полифенолов показано, что эфирная и этилацетатная вытяжки, в которых наблюдается относительно большое содержание фенольных соединений, обладают более выраженными антимикробной активностью против грамотрицательных и грамположительных бактерий и антирадикальной активностью in vitro [6]. Имеются данные по фармакокинетическому исследованию ингибирующего влияния полисахаридов сапожниковии растопыренной корней на процесс разрушения хромонов в кишечном тракте [7]. ...
The aim of this work was to determine the total sum of flavonoids, tannins and the quantitative content of polysaccharide fractions
of Saposhnikoviae divaricatae radices collected in Russia, Mongolia and obtained in China.
Plant material on different phases of development was collected during field works in Tarbagataisky, Mukhorshibirsky, Ivolginsky
and Kyakhtinsky districts of the Republic of Buryatia (Russia), in Aginsky district of the Transbaikal Territory (Russia) and in Khentei
aimak (Mongolia), and obtained in a pharmacy store in Xining, Qinghai Province (China).
The total sum of flavonoids was determined by spectrophotometric method. The content of tannins was determined by permanganatometric method in terms of tannin. The quantitative content of polysaccharide fractions was carried out by gravimetric method.
The total sum of flavonoids in terms of rutin in Saposhnikoviae divaricatae radices was 0.29−0.48%. The quantitative content of
tannins was ranged from 0.38 to 0.55%. Water-soluble polysaccharides were represented by an amorphous yellowish-gray powder, soluble in water. The content ranged from 6.8 to 8.3%. The fraction of pectin substances was represented by an amorphous gray
powder, soluble in water. The quantitative content of the sample from Tarbagataisky region was the highest − 1.1%, while the amount
of the rest ranged from 0.3 to 0.4%. The quantitative content of hemicellulose A in studied samples varied significantly, and ranged
from 2.5% to 13.8%. The yield of grayish-brown powder of hemicellulose B was 1.8-14.3%.
Conclusion. For the first time the content of the total sum of flavonoids, tannins and polysaccharides in the roots of Saposhnikovia
divaricata from Russian and Mongolian flora was studied in this research. It was found that this species accumulates significant
quantity of flavonoids and tannins along with coumarins and chromones. Relatively high content of water-soluble polysaccharides in S. divaricata shows promising studying of their composition and structure.
... Schischk. of the family Umbelliferae. It has been widely used as a traditional medicine for alleviating inflammation, rheumatic conditions, spasms, and other ailments in East Asia (3)(4)(5)(6)(7). Numerous active compounds with pharmaceutical potential have been identified in Saposhnikoviae Radix, including chromones (prim-O-glucosylcimifugin, 4'-O-β-d-glucosyl-5-O-methylvisamminol, 3'-O-angeloylhamaudol, sec-O-glucosylhamaudol, and cimifugin), and coumarins (nodakenin, psoralen, bergapten, imperatorin, phellopterin, and anomalin), among others (4,8,9). ...
Although the dried root of Saposhnikovia divaricata (Turcz.) Schischk. (Umbelliferae) is a popular medicinal plant in East Asia, there has been no systemic toxicological evaluation of a water extract of Saposhnikoviae Radix (SRE). In this experiment, an oral acute and 13-week subchronic toxicological evaluations of SRE (500-5,000 mg/ kg body weight) were performed in both sexes of Crl:CD(SD) rats. Based on the results from mortality, clinical signs, effects on body weight and organ weight, clinical biochemistry, hematology, urinalysis, and histopathology, significant acute, 4-week repeated dose range finding (DRF) and 13-week subchronic toxicity of SRE was not observed in either sex of rats; thus, the no observed adverse effect level (NOAEL) was 5,000 mg (kg/day). To identify anti-hyperuricemia potential of SRE, the suppressive effect of SRE was determined in mice challenged with potassium oxonate (PO; 250 mg/kg) via intraperitoneal injection for 8 days (each group; n = 7). SRE supplementation suppressed the uric acid level in urine through significant xanthine oxidase (XO) inhibitory activity. Kidney dysfunctions were observed in PO-challenged mice as evidenced by an increase in serum creatinine level. Whereas, SRE supplementation suppressed it in a dose-dependent manner. Collectively, SRE was safe up to 5,000 mg (kg/day) based on NOAEL found from acute and 13-week subchronic toxicological evaluations. SRE had anti-hyperuricemia effect and lowered the excessive level of uric acid, a potential factor for gout and kidney failure.
... In recent years, the pharmacological activity and chemical constituents of RS have been extensively studied by researchers worldwide. Pharmacological studies have shown that the ethanol extract of RS has anti-inflammatory [2][3][4], antioxidant [5][6][7], analgesic [8], antipyretic [9], anticancer [10], and antiviral effects [11]. RS mainly contains chromones [12], volatile oils, coumarins [13], polysaccharides [14], and organic acids, with chromones being the main active components. ...
A rapid and sensitive quantitative analytical method was established for the simultaneous determination of five chromones (prim-O-glucosylcimifugin, cimifugin, 4′-O- β -D-glucosyl-5-O-methylvisamminol, 5-O-methylvisammiol, and sec-o-glucosylhamaudol) in the plasma of RS-treated rats for the first time using ultra performance liquid chromatography- (UPLC-) tandem mass spectrometry. The Waters Acquity UPLC BEH C18 (50 mm × 2.1 mm, 1.7 μ m) was used as the chromatographic column, 0.1% formic acid water and 0.1% formic acid acetonitrile comprised the mobile phases, and all samples were determined under positive ion mode. The results showed that all analytes had good linearity ( r>0.9902 ), between-day and within-day precisions less than 15%, accuracy between −5.50% and 5.53%, and extraction recovery between 88.26% and 97.65%. Both the matrix effect and stability met the requirements. This method was successfully applied for the comparative pharmacokinetics of five active components of RS in normal and febrile rats. The results showed that the pharmacokinetic behavior of RS extract significantly differed between the two types of rats.
... Schischk belongs to the Apiaceae family, and is a traditional Chinese herb dating back to Qin-Han dynasty. It is mainly cultivated in the northern and northeastern regions of China (Kreiner et al. 2017), and is widely found and studied in Japan and Korea (Tal and Cheung 2007;Khan and Kim 2013). The diverse pharmacological effects of the herb include antipyretic, analgesic, antiinflammatory and anti-arthritic effects (Wang et al. 1999). ...
Saposhnikovia divaricata (Turcz.) Schischk is a traditional herb of East Asia. Bioactive chromones and volatile components in its roots are known to exhibit pharmacological functions. However, limited information is available on the drought resistance of this herb. In this study, potted Saposhnikovia divaricata seedlings were subjected to a progressive drought stress of 20 days by withholding water followed by twice rehydration, which resulted in some physiological, biochemical and secondary metabolite responses as well as drought acclimatization. A decline in leaf water content but increase in electrolyte leakage, malondialdehyde (MDA), hydrogen peroxide (H2O2), glutathione (GSH), proline, soluble sugar, prim-O-glucosylcimifugin and 4′-O-β-d-glucosyl-5-O-methylvisamminol content was observed. After rehydration, some of the indices recovered except proline, soluble sugar, prim-O-glucosylcimifugin and 4′-O-β-d-glucosyl-5-O-methylvisamminol content. Moreover, mild (day 8), moderate (days 12–16) and severe (day 20) drought phases were identified. A total of 18 volatile components were identified by GC–MS under different drought phases, of which aromatic alcohols (42.02%) and sesquiterpenes (37.35%) were the major components. The characteristic component named falcarinol was decreased by severe drought stress. This study demonstrated that Saposhnikovia divaricata had strong drought acclimatization, and resisted drought by activating the antioxidant system and accumulating osmolytes. In addition, moderate and severe drought stress promoted bioactive secondary metabolites prim-O-glucosylcimifugin and 4′-O-β-d-glucosyl-5-O-methylvisamminol accumulation. Severe drought stress reduced falcarinol relative content, which provided an insight for improving the quantity of Saposhnikovia divaricata bioactive components.
... Schischk. [1]), Radix Astragali (RA; Huangqi; the roots of Astragalus mongholicus Bunge [1,2]) and Rhizoma Atractylodis macrocephalae (RAM; Baizhu; the rhizomes of Atractylodes 264.7 murine macrophages [19]. More than 100 compounds, such as isoflavonoids, triterpene saponins, polysaccharides, and amino acids have been identified in RA so far [20,21]. ...
Yu Ping Feng San (YPFS) is a classical TCM formulation which has been traditionally used for treatment of immune system related diseases such as chronic bronchitis, allergic rhinitis and asthma. The formula is a mixture of Radix Saposhnikoviae (Fangfeng), Radix Astragali (Huangqi), and Rhizoma Atractylodis macrocephalae (Baizhu). TLC- and LC-DAD-ESI-MS/MS methods have been developed for the analysis of the metabolic profiles of the single herbs and of the formula. Decoctions and ASE extracts were analysed in order to trace components of the individual herbs in YPFS. Nine constituents of Radix Saposhnikoviae, ten constituents of Radix Astragali and five constituents of Rhizoma Atractylodis macrocephalae have been assigned in the chemical profiles of the formula, which now allow the standardization of YPFS. The pharmacological testing showed that all extracts significantly inhibited expression of TNF-α, IFN-γ, and IL-1β in U937 cells, while the inhibition of IL-4 was consistently low. Compared to conventional analyses which are focused on a limited set of compounds, metabolomics approaches, together with novel data processing tools, enable a more holistic comparison of the herbal extracts. In order to identify the constituents which are relevant for the immunomodulatory effects of the formula, metabolomics studies (PCA, OPLS-DA) have been performed using UPLC/QTOF MS data.
... Hiroe] (Apiaceae) inhabits Mongolia, China, Korea, eastern Siberia (Republic of Buryatia, Zabaikalsky Krai) and the Russian Far East (Sakhalin Oblast, Priamurye, Primorye) and has been used for several thousand years in traditional medicine of China, Japan, and Korea [1]. It possesses a broad spectrum of pharmacological activity including anticarcinogenic and neuroprotective [2][3][4][5]. ...
... (Radix Saposhnikoviae) has been used for many years in China as an important herbal medicine and is officially listed in the pharmacopoeia of the People's Republic of China [19]. Radix Saposhnikoviae has been shown to have activities against various human diseases and conditions, including antipyretic, analgesia [20,21], antioxidation [22,23], anti-inflammatory [20,24], inhibition of platelet aggregation and anticlotting [21]. Chromones were determined to be effective ingredients of the species, and among the chromones, prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside, and sec-O-glucosylhamaudol are representative constituents of Radix Saposhnikoviae that are often used for quality control of Radix Saposhnikoviae [25,26]. ...
Ionic liquid-based ultrasonic-assisted extraction (ILUAE) was successfully applied to the extraction of the four chromones (prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside, and sec-O-glucosylhamaudol) from Saposhnikovia divaricata (Radix Saposhnikoviae) for the first time. A series of l-alkyl-3-methylimidazolium ILs differing in anion and cation compositions was evaluated for extraction efficiency, and [C3MIM]Br was selected as the optimal solvent. In addition, ultrasound extraction parameters were optimized, and the chromones were directly quantified and analyzed by rapid resolution liquid chromatography-electrospray ionization/mass spectrometry (RRLC-ESI/MS). The optimal conditions were as follows: 0.4 M concentration of [C3MIM]Br, 20:1 solvent to solid ratio, and ultrasonic time, temperature, and frequency of 5 min, 40 °C, and 50 kHz, respectively. This approach obtained the highest extraction yield of 10.188 ± 0.473 mg g-1 for total chromones. Compared with regular UAE, the proposed approach exhibited a higher efficiency (61.56% increase) and shorter extraction time (nine times shorter). Also, ILUAE was an efficient, rapid, and simple sample preparation technique for extraction of chromones, and the established RRLC-DAD method could serve as a rapid and effective technique for extracting chromones from Radix Saposhnikoviae.
... Prim-O-glucosylcimifugin, the chromone with the highest content in the roots of SD, showed significant anti-inflammatory effects on LPS-induced inflammatory responses in RAW 264.7 cells and significantly protected mice against LPS-induced acute lung injury [25]. SD ethanol extract and chromones isolated from SD showed potential anti-inflammatory and protective effects in LPSactivated RAW 264.7 cells [11,26]. In the present study, we confirmed the presence of the chromones, which were prim-O-glucosylcimifugin, 4 -O-glucosyl methylvisamminol, and sec-O-glucosylhamaudol in SDE, and this partially explains the anti-inflammatory activity of SDE. ...
... Prim-O-glucosylcimifugin, the chromone with the highest content in the roots of SD, showed significant anti-inflammatory effects on LPSinduced inflammatory responses in RAW 264.7 cells and significantly protected mice against LPS-induced acute lung injury [25]. SD ethanol extract and chromones isolated from SD showed potential anti-inflammatory and protective effects in LPS-activated RAW 264.7 cells [11,26]. In the present study, we confirmed the presence of the chromones, which were prim-O-glucosylcimifugin, 4 -O--D-glucosyl-5-O-methylvisamminol, and sec-O-glucosylhamaudol in SDE, and this partially explains the anti-inflammatory activity of SDE. ...
Saposhnikovia divaricata
Schischkin has been used in traditional medicine to treat pain, inflammation, and arthritis. The aim of this study was to investigate the anti-inflammatory and antiosteoarthritis activities of
Saposhnikovia divaricata
extract (SDE). The anti-inflammatory effect of SDE was evaluated
in vitro
in lipopolysaccharide- (LPS-) treated RAW 264.7 cells. The antiosteoarthritic effect of SDE was investigated in an
in vivo
rat model of monosodium iodoacetate- (MIA-) induced osteoarthritis (OA) in which rats were treated orally with SDE (200 mg/kg) for 28 days. The effects of SDE were assessed
in vivo
by histopathological analysis and by measuring weight-bearing distribution, cytokine serum levels, and joint tissue inflammation-related gene expression. SDE showed anti-inflammatory activity by inhibiting the production of nitric oxide (NO), prostaglandin E
2
(PGE
2
), tumor necrosis factor-
α
(TNF-
α
), and interleukin-6 (IL-6) in LPS-induced RAW 264.7 cells. In addition, SDE promoted recovery of hind limb weight-bearing, inhibited the production of proinflammatory cytokines and mediators, and protected cartilage and subchondral bone tissue in the OA rat model. Therefore, SDE is a potential therapeutic agent for OA and/or associated symptoms.
... Schizonepeta tenuifolia has a wide range of the physiological activities against inflammationrelated diseases and oxidative stress [56,57,58]. Saposhnikovia divaricata is used to treat rheumatoid arthritis via inhibiting NF-κB and MARKs [59], and has potent antioxidant, antiinflammatory, and protective properties on lipopolysaccharide-activated RAW 264.7 cells [60]. Poria cocos has been traditionally used in Oriental medicine as a diuretic and sedative [61] and 25-methoxyporicoic acid A and other triterpene acids from Poria cocos have anti-tumor-promoting effects [62]. ...
The preventive and therapeutic mechanisms in multiple sclerosis are not clearly understood. We investigated whether Hyungbangpaedok-san (HBPDS), a traditional herbal medicine, has a beneficial effect in experimental autoimmune encephalomyelitis (EAE) mice immunized with myelin oligodendrocyte glycoprotein peptide (MOG35-55). Onset-treatment with 4 types of HBPDS (extracted using distilled water and 30%/70%/100% ethanol as the solvent) alleviated neurological signs, and HBPDS extracted within 30% ethanol (henceforth called HBPDS) was more effective. Onset-treatment with HBPDS reduced demyelination and the recruitment/infiltration and activation of microglia/macrophages in the spinal cord of EAE mice, which corresponded to the reduced mRNA expression of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β), iNOS, and chemokines (MCP-1, MIP-1α, and RANTES) in the spinal cord. Onset-treatment with HBPDS inhibited changes in the components of the blood-brain barrier such as astrocytes, adhesion molecules (ICAM-1 and VCAM-1), and junctional molecules (claudin-3, claudin-5, and zona occludens-1) in the spinal cord of EAE mice. Onset-treatment with HBPDS reduced the elevated population of CD4+, CD4+/IFN-γ+, and CD4+/IL-17+ T cells in the spinal cord of EAE mice but it further increased the elevated population of CD4+/CD25+/Foxp3+ and CD4+/Foxp3+/Helios+ T cells. Pre-, onset-, post-, but not peak-treatment, with HBPDS had a beneficial effect on behavioral impairment in EAE mice. Taken together, HBPDS could alleviate the development/progression of EAE by regulating the recruitment/infiltration and activation of microglia and peripheral immune cells (macrophages, Th1, Th17, and Treg cells) in the spinal cord. These findings could help to develop protective strategies using HBPDS in the treatment of autoimmune disorders including multiple sclerosis.
... In addition, induction of apoptosis in the activated cancer cells may be an effective strategic approach for cancer therapy. Moreover, some of recent studies reported that several traditional herbal medicine extracted from herbal plants as Sho-saiko-to, Saposhnikovia divaricata and Salvia miltiorrhiza Bunge induced apoptosis and cell cycle arrest in HL-60 cell line (Sung et al., 1999;Makino et al., 2006;Tai and Cheung, 2007). ...
Chios gum mastic (CGM) is produced from Pistiacia lentiscus L var chia, which grows only on Chios Island in Greece. CGM is a kind of resin extracted from the stem and leaves, has been used for many centuries in many Mediterranean countries as a dietary supplement and folk medicine for stomach and duodenal ulcers. CGM is known to induce cell cycle arrest and apoptosis in some cancer cells. This study was undertaken to investigate the alteration of the cell cycle and induction of apoptosis following CGM treatment of HL-60 cells. The viability of the HL-60 cells was assessed using the MTT assay. Hoechst staining and DNA electrophoresis were employed to detect HL-60 cells under-going apoptosis. Western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, MMP activity and proteasome activity analyses were also employed. CGM treatment of HL-60 cells was found to result in a dose-and time-dependent decrease in cell viability and apoptotic cell death. Tested HL-60 cells showed a variety of apoptotic manifestations and induced the downregulation of G1 cell cycle-related proteins. Taken collectively, our present findings demonstrate that CGM strongly induces G1 cell cycle arrest via the modulation of cell cycle-related proteins, and also apoptosis via proteasome, mitochondrial and caspase cascades in HL-60 cells. Hence, we provide evidence that a natural product, CGM could be considered as a novel therapeutic for human leukemia.
... Saposhnikoviae radix is used for affections of skin and mucosa [24][25][26]. It was found to inhibit NO production through iNOS and its mRNA expression in LPS-induced RAW 264.7 cells [107]. ...
Oral mucositis is one of the common side effects of chemotherapy treatment with potentially severe implications. Despite several treatment approaches by conventional and complementary western medicine, the therapeutic outcome is often not satisfactory. Traditional Chinese Medicine (TCM) offers empirical herbal formulas for the treatment of oral ulceration which are used in adaptation to chemotherapy-induced mucositis. While standard concepts for TCM treatment do not exist and acceptance by conventional oncologists is still low, we conducted a review to examine the evidence of Chinese herbal treatment in oral mucositis. Eighteen relevant studies on 4 single herbs, 2 combinations of 2 herbs, and 11 multiherbal prescriptions involving 3 or more compounds were included. Corresponding molecular mechanisms were investigated. The knowledge about detailed herbal mechanisms, especially in multi-herbal prescriptions is still limited. The quality of clinical trials needs further improvement. Meta-analysis on the existent database is not possible but molecular findings on Chinese medicinal herbs indicate that further research is still promising for the treatment of chemotherapy-induced oral mucositis.
Ethnopharmacological relevance:
Saposhnikoviae Radix (SR) is a traditional Chinese medicine, known as "Fangfeng". As one of the main active components, Saposhnikoviae Radix polysaccharides (SP) has demonstrated a range of biological activities, especially immunity regulation activity.
Aims of the study:
This study aimed at exploring whether polysaccharides have activity after degradation, then discovering the potential effective fragments of SP.
Materials and methods:
Here we establish the chromatographic fingerprints method for 32 batches of 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatives of oligosaccharides by HPLC, meanwhile evaluating its immunomodulatory activity in vivo. Then, the potential effective fragments of SP were screened out based on the spectrum-effect relationship analysis between fingerprints and the pharmacological results. Besides, liquid chromatography ion trap-time of flight mass spectrometry (LC-IT-TOF MS) coupled with multiple data-mining techniques was used to identify the potential effective oligosaccharides.
Results:
These findings showed that the hydrolysate of SP have significant immunomodulatory, and the immunity regulation activity varies under different hydrolysis conditions. The 4 potential effective peaks of the hydrolysate of SP were mined by spectrum-effect relationship. Finally, the chemical structure of 4 potential effective oligosaccharide fragments of SP was elucidated based on LC-IT-TOF MS. F10 was inferred tentatively to be Hex1→6Hex1→6Hex1→6Hex1→6Hex1→6Gal; F18 was confirmed to be Rhamnose; F14 was inferred tentatively to be Hex1→4Hex1→ 4Hex1→4Gal and F25 was tentatively inferred to be Ara1→6Gal.
Conclusions:
This study may provide a sound experimental foundation in the exploration of the active fragments from macromolecular components with relatively complex structures such as polysaccharides.
Ethnopharmacological relevance:
The incidence of allergic disease is constantly increasing, but its pathogenesis is not fully understood. Saposhnikovia divaricata (SD), called 'Fangfeng' in China, not only can be used for antipyretic, analgesic and anti-inflammatory as a traditional Chinese medicine, but also as an active ingredient in about 8% prescriptions. However, its effects on type I allergy and pseudoallergy have not been clarified.
Aim of the study:
To explore the treatment and potential mechanisms of SD and its major bioactive component Prim-O-glucosylcimifugin (POG) on type I allergy and pseudoallergy in vitro and in vivo.
Materials and methods:
The inhibitory effect of SD decoction and POG on type I allergy and its possible mechanism were evaluated by using RBL-2H3 cells model in vitro and the passive cutaneous anaphylaxis (PCA) mouse model in vivo. The cell degranulation of RBL-2H3 cells induced by DNP-IgE/DNP-BSA and Compound 48/80 (C48/80) was investigated, and the molecules of degranulation related signaling pathway was further detected by qRT-PCR and Western Blot analysis. Meanwhile, therapeutic effect of SD Decoction and POG were evaluated using PCA models in vivo. The molecular docking technology was conducted to explore the potential mechanisms.
Results:
In cells model induced by DNP-IgE/DNP-BSA, the release rate of β-Hex in high dose of SD and POG groups were 43.79% and 57.01%, and the release amount of HA in high dose of SD and POG groups were 26.19 ng/mL and 24.20 ng/mL. They were significantly lower than that in the model group. Besides, SD decoction and POG could significantly inhibit intracellular Ca2+ increasing and cell apoptosis. But there is no obvious effect on cells degranulation induced by C48/80. The molecular docking results showed that 5-O-Methylvisamioside and POG could bind with FcεRI α with stronger binding ability, but weak binding ability to Mrgprx2. Moreover, qPCR and Western blot analyses indicated that SD could down-regulate Lyn/Syk/PLCγ, MAPK and PI3K/AKT/NF-κB signal pathway to inhibit IgE-dependent cell degranulation. In mice PCA model, both SD and POG could dose-dependently attenuate the Evans Blue extravasation, paw and ear swelling induced by DNP-IgE/DNP-BSA, but no significant inhibition under the PCA models induced by C48/80.
Conclusion:
In conclusion, SD is effective for the therapeutic of type I allergies, suggesting that SD is a potential candidate for the treatment of type I allergy, and the underlying mechanism of these effects needs to be further studied.
The dried root of Saposhnikovia divaricata (Turcz.) Schischk. is popular as a good medicinal material, however the abundant aerial part is often discarded, which caused the waste of resources. In order to exploit resources, the essential oils of the plant aerial part and root were extracted, separately called as VOA and VOR, their chemicals were identified. The tumor necrosis factor-α, interleukin-6, nitric oxide and interleukin-1β were detected to evaluate the oils anti-inflammatory activities. Then, the oils free radical scavenging rates were measured with DPPH, ABTS and hydroxyl free radical. The oils antitumor activities were evaluated with HeLa and HCT-8 cancer cell lines. The results showed the concentrations of VOA and VOR were separately 0.261% and 0.475%. Seventeen components of VOA were identified, accounting for 80.48% of VOA, including phytol, spathulenol, phytone, 4(15),5,10(14)-Germacratrien-1-ol, neophytadiene, etc. Seven components of VOR were determined, representing 90.73% of VOR, consisted of panaxynol, β-bisabolene, etc. VOA and VOR significantly inhibited the secretion of nitric oxide, interleukin-1β, interleukin‐6 and tumor necrosis factor-α, effectively scavenged the DPPH, ABTS and hydroxyl free radicals, and showed significant antiproliferative activity against HeLa and HCT-8. The two oils presented important biological activity, which provided a hopeful utilized basis, and helped to reduce the waste of the aerial non-medicinal resources of S. divaricata.
Ethnopharmacological relevance:
Ulcerative colitis (UC) has become a healthy burden worldwide due to its insidious onset and repetitive relapse, with a rather complex etiology, including inappropriate immune response, dysbiosis, genetic susceptibility, and unhealthy diets. The Wu-Shi-Cha (WSC) formula is a widely utilized drug to protect against gastrointestinal disorders.
Aim of the study:
The study aspired to dissect the pertinent mechanisms of the WSC to treat UC.
Materials and methods:
Network pharmacology and weighted gene co-expression network analysis (WGCNA) were performed to predict the targets of WSC in the context of UC and colorectal cancer. Dextran sodium sulfate (DSS) was used to construct murine models of experimental colitis, and the WSC was given to colitis mice for 14 days. Feces and colon samples were subjected to 16S rRNA gene sequencing combined with liquid chromatography-mass spectrometry (LC-MS) and biochemical experiments, respectively.
Results:
Network pharmacology analysis predicted that the WSC formula could orchestrate inflammation, infection, and tumorigenesis, and WGCNA based on The Cancer Genome Atlas (TCGA) database showed a potent anti-neoplastic effect of the WSC therapy for colorectal cancer. The WSC therapy rescued bursts of pro-inflammatory cytokines and colonic epithelial collapse in DSS-induced colitis mice. Moreover, the high dose of WSC treatment facilitated the alternative activation of peritoneal macrophages (Mφs) and these Mφs were conducive to the survival of intestinal stem cells (ISCs), and the disturbed homeostasis of gut microbiota was re-established after WSC treatment, as evidenced by the decreased colonization of pathological taxa in the fecal samples.
Conclusion:
The WSC formula suppresses inflammation and re-establishes the homeostasis of gut microbiota, thereby ameliorating colitis progression.
Low-temperature stratification has a beneficial effect on releasing seed dormancy and germination. Taking Saposhnikovia divaricata (Trucz.) Schischk. seeds as the research object, the effects of low-temperature stratification on dormancy release and germination of S. divaricata seeds were studied. The morphological, physiological and biochemical indexes, hormone content and transcriptome of dormant seeds, seeds in the dormancy release period and germination period were discussed. The results showed that 4 °C low-temperature stratification treatment increased the contents of soluble sugar, gibberellin and indoleacetic acid, enhanced amylase and α-amylase activity, but decreased the contents of starch and abscisic acid. A total of 100,690 unigenes were obtained from S. divaricata transcriptome. RNA-seq analysis showed that compared with dormant seeds (CK), the number of up-regulated genes in seeds in the dormancy release period (S) and germination period (G) was 13,998 and 16,817, and the number of down-regulated genes was 9237 and 14,054, respectively. This study identified a large number of differentially expressed genes related to plant hormone signal transduction, starch and sucrose, plant energy metabolism pathway and hormones in low-temperature stratification reaction. The transcriptomic gene expression profiles present a valuable genomic tool to the molecular mechanisms of S. divaricata seed germination underlying low-temperature stratification, which can provide theoretical basis and practical guidance for cultivation and planting of S. divaricata.
The perennial herb Saposhnikovia divaricata SCHISCHKIN, used as a source of traditional medicines, is known for its anti-bone erosion, anti-inflammatory, and analgesic effects. We examined the osteogenic properties of Sec-O-glucosylhamaudol (SOGH) extracted from S. divaricata roots as a bone-protective compound. We found that SOGH promoted osteoblast differentiation and mineralization and induced the expression of Bsp, Ocn, and Opn without any cytotoxic effects. SOGH also increased nuclear RUNX2 levels in pre-osteoblasts via the BMP2 and Wnt3a signaling pathways, but it had no evident effects on the autophagic and necroptotic signaling pathways. Furthermore, SOGH promoted bone-forming activities by inducing the adhesion and migration of osteoblasts and the expression of RUNX2 target proteins, including VEGF and MMP13. In sum, SOGH has osteogenic properties mediated via the BMP2 and Wnt3a signaling pathways and RUNX2 expression, indicating the bone-protective potential of this compound in diseases such as periodontitis and osteoporosis.
One new chromone, (2′S,4′S)-5′-hydroxy-5-O-methylvisamminol (1), was isolated from the roots of Saposhnikovia divaricata. The structure of compound 1 was identified by spectroscopic methods including 1D, 2D NMR, and HR-ESI-MS data. The absolute configuration of compound 1 was elucidated by electronic circular dichroism (ECD) calculation. Compound 1 showed moderate cytotoxic activity against human prostatic cancer PC-3 and breast cancer MCF-7 cell lines, with IC50 values of 52.66 ± 1.73 and 83.12 ± 8.50 μM, respectively.
Aims
The current study aims to investigate the effect of Yupingfeng (YPF) powder on immunosuppression, and explore the possible mechanisms.
Main methods
Firstly, the monomer components of YPF powder were analyzed by UPLC-QTOF-MS combined with UNIFI automatic analysis platform, then the mechanism of YPF on immunosuppressive treatment was investigated using network pharmacological method, and finally the prediction was verified in a Candida albicans (Can)-induced immunosuppressive BALB/c mouse model.
Key findings
98 monomer compounds in YPF were obtained. Through virtual analysis and screening on the oral utilization and drug likeness properties of the components, 47 effective components were got. 9 core targets obtained were enriched in IL-17 signaling pathway. In the mouse model, YPF could reduce the number of Can and alleviate Can-induced inflammation in the kidney effectively, upregulate Can-induced low proportion of CD4⁺/CD8⁺ of splenic lymphocytes, and increase Can-induced low activity of IL-17 pathway.
Significance
These results demonstrate that YPF could improve the immunity of Can-induced immunosuppression in BALB/c mice through upregulating the activity of IL-17 pathway.
Prim‐O‐glucosylcimifugin, cimifugin and 5‐O‐methylvisamminoside are three major chromone derivatives of Saposhnikovia divaricata that have many pharmacological activities, such as anti‐inflammatory and antitumor activities. In the present work, an effective method for the simultaneous separation of prim‐O‐glucosylcimifugin, cimifugin and 5‐O‐methylvisamminoside with high purities was established using HPD‐300 resin coupled with preparative high‐performance liquid chromatography. The adsorption kinetics curves of the three compounds on the HPD‐300 resin were studied and found to fit well according to the pseudo‐second‐order equation. The adsorption isotherm results indicated that the adsorption process of the three compounds was exothermic. After a one‐run treatment with the resin, the contents of prim‐O‐glucosylcimifugin, cimifugin and 5‐O‐methylvisamminoside increased from 0.29%, 0.06% and 0.37% to 13.07%, 2.83% and 16.91% with recovery yields of 76.38%, 78.25% and 76.73%, respectively. Finally, the purities of the three compounds were found to reach more than 95% after further separation using preparative high‐performance liquid chromatography. The method developed in this study was effective and could simultaneously separate three chromones from Saposhnikovia divaricate. The experimental results also showed that the HPD‐300 resin is suitable for the separation of chromone derivatives. This article is protected by copyright. All rights reserved
Four new polyacetylene substances, sadivaethynes A–D, were isolated from the ethanol extract of the roots of Saposhnikovia divaricata (Turcz.) Schischk using repeated column chromatography. Structural elucidation of compounds 1–4 was established by 1D and 2D NMR spectra referring to the literature, together with high-resolution mass spectrometric analysis. All compounds were evaluated for cytotoxicity against two human cancer cell lines (MGC-803, Ishikawa) in vitro.
Natural products are favored because of their non-toxicity, low irritants, and market reacceptance. We collected examples, according to ancient wisdom, of natural products to be applied in transdermal delivery. A transdermal delivery system, including different types of agents, such as ointments, patches, and gels, has long been used for skin concerns. In recent years, many novel transdermal applications, such as nanoemulsions, liposomes, lipid nanoparticles, and microneedles, have been reported. Nanosized drug delivery systems are widely applied in natural product deliveries. Nanosized materials notably enhance bioavailability and solubility, and are reported to improve the transdermal permeation of many substances compared with conventional topical formulations. Natural products have been made into nanosized biomaterials in order to enhance the penetration effect. Before introducing the novel transdermal applications of natural products, we present traditional methods within this article. The descriptions of novel transdermal applications are classified into three parts: liposomes, emulsions, and lipid nanoparticles. Each section describes cases that are related to promising natural product transdermal use. Finally, we summarize the outcomes of various studies on novel transdermal agents applied to skin treatments.
Background:
5-O-methylvisammioside (MeV), also known as 4'-O-β-D-glucosyl-5-O-methylvisamminol, is a conventional marker compound for quality control of roots of Saposhnikovia diviaricata (Radix Saposhnikoviae), which exhibits anti-inflammatory and neuroprotective activities.
Purpose:
According to the activity of MeV, we speculated that MeV may have antidepressant effect on LPS induced depression, and further explored its mechanism.
Study design:
First, to explore the effect and mechanism of MeV on LPS-induced depression in mice, and then to further explore the effect and mechanism of MeV on LPS-activated BV-2 microglia.
Methods:
By the OFT, EPM, TST and FST behavioral tests, to explore the effect of MeV pretreatment on the behavior of LPS-induced depression mice. ELISA and Griess method were used to detect the changes of the serum TNF-α and IL-6 levels, the hippocampus SOD and MDA levels, and the NO, SOD, MDA, TNF-α and IL-6 levels in the culture medium of LPS-stimulated BV-2 microglia. Western blot was used to analyze the protein expression in the Nf-κB/IκB-α and BDNF/TrkB pathway in the hippocampus of mice and BV-2 microglia.
Results:
MeV (4 mg/kg, i.p.) pretreatment significantly improves the activity and exploration ability of LPS-induced depression mice, and reduces the immobility time. MeV inhibited the production of pro-inflammatory cytokines in the serum of mice induced by LPS, such as IL-6 and TNF-α. MeV also increased the levels of SOD and reduces the expression of MDA in the hippocampus, thus promoting the alleviation of depressive symptoms in mice. Western blotting analysis showed that the antidepressant activity of MeV was related to the decrease of Nf-κB nuclear transport, the inhibition of IκB-α phosphorylation, and the increase of BDNF and TrkB expression. MeV (40 μM) significantly reduced the contents of NO, MDA, TNF-α and IL-6 in the culture medium of LPS-stimulated BV-2 microglia, and increased the content of SOD.
Conclusion:
MeV can regulate the neurotrophic factors in the mouse brain, reduce the content of inflammatory factors by the Nf-κB/IκB-α pathway, improve oxidative stress, and inhibit the excessive activation of LPS-stimulated BV -2 microglia. It effectively reversed the depression-like behAavior induced by LPS in mice.
Previously, we established a 1H NMR metabolomics method using reversed-phase solid-phase extraction column (RP-SPEC), and succeeded in distinguishing wild from cultivated samples of Saposhnikoviae radix (SR), and between SR and its substitute, Peucedanum ledebourielloides root (PR). Herein, we performed LC-HR/MS metabolomics using fractions obtained via RP-SPEC to identify characteristic components of SR and PR. One and three characteristic components were respectively found for SR and PR; these components were isolated with their m/z values and retention times as a guide. The characteristic component of SR was identified as 4′-O-β-d-glucosyl-5-O-methylvisamminol (1), an indicator component used to identify SR in the Japanese Pharmacopoeia. In contrast, the characteristic components of PR were identified as xanthalin (2), 4′-O-β-d-apiosyl (1 → 6)-β-d-glucosyl-5-O-methylvisamminol (3), and 3′-O-β-d-apiosyl (1 → 6)-β-d-glucosylhamaudol (4) based on spectroscopic data such as 1D- and 2D-NMR, MS, and specific optical rotation. Among them, 4 is a novel compound. For the correlation between the NMR metabolomics results in the present and our previous report, only 1 and 2 were found to correlate with the chemical shifts, and the other compounds had no correlation. As the chemical shifts for compounds 1, 3, and 4 were similar to each other, especially for the aglycone moiety, they could not be distinguished because of the sensitivity and resolution of 1H NMR. Accordingly, combining NMR and LC/MS metabolomics with their different advantages is considered useful for metabolomics of natural products. The series of methods used in our reports could aid in quality evaluations of natural products and surveying of marker components.
We investigated the mechanism of action of panaxynol (PAL) extract from the root of Saposhnikovia diviaricata (Turcz.) Schischk for treating acute liver injury caused by lipopolysaccharide (LPS) and D-galactosamine (D-Gal N) in mice. A mouse model of acute liver failure induced by LPS/D-Gal N was established. Mice were divided randomly into three equal groups: control group, LPS/D-Gal N group and PAL group. After seven days of continuous PAL administration, all animals except controls were injected with 50 μg/kg LPS and 800 mg/kg D-Gal N; blood and liver samples were collected after 8 h. Compared to the LPS/D-Gal N group, the levels of catalase, glutathione and superoxide dismutase were increased in the liver of the PAL group. The inflammatory response index indicated that PAL attenuated LPS/D Gal N-induced liver pathological injury and decreased levels of hepatic malondialdehyde, serum alanine aminotransferase, aspartate transaminase, tumor necrosis factor-α, and interleukins 1β and 6. PAL also inhibited LPS/D-Gal N induced nuclear factor-kappa B (Nf-κB), inhibitor kappa B-α (IκB-α) activation, and up-regulated Nrf2 and heme oxygenase-1 (HO-1) expression. PAL can prevent LPS/D-Gal N induced acute liver injury by activating Nrf2/HO-1 to stimulate antioxidant defense and inhibit the IkB-α/NF-κB signaling pathway.
H NMR-based metabolomics has been applied in research on food, herbal medicine, and natural products. Although excellent results were reported, samples were directly extracted with a deuterated solvent (e.g., methanol-d4 or D2O) in most reports. As primary metabolites account for most of the results, data for secondary metabolites are partially reflected. Consequently, secondary metabolites tend to be excluded from factor loading analysis, serving as a significant unfavorable feature of ¹H NMR-based metabolomics when investigating biologically active or functional components in natural products and health foods. Reversed-phase solid-phase extraction column (RP-SPEC) was applied for sample preparation in ¹H NMR-based metabolomics to overcome this feature. The methanol extract from Saposhnikoviae radix (SR), an important crude drug, was fractionated with RP-SPEC into 5% methanol-eluting fractions, and the remaining fraction was collected. Each fraction was subjected to ¹H NMR-based metabolomics and compared to results from conventional ¹H NMR-based metabolomics. Based on principal component analysis (PCA) and partial least squares projections to latent structures discriminant analysis (PLS-DA), the 5% methanol fraction and conventional method reflected the amount of saccharides such as sucrose on the PC1/PLS1 axes, and wild and cultivated samples were discriminated along those axes. The remaining fraction clearly distinguished SR from Peucedanum ledebourielloides root. The compounds responsible for this discrimination were deemed falcarindiol derivatives and other unidentified secondary metabolites from the s-plot on PLS-DA. The secondary metabolites from original plants were, therefore, presumed to be concentrated in the remaining fraction by RP-SPEC treatment and strongly reflected the species differences. The developed series is considered effective to perform quality evaluation of crude drugs and natural products.
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Saposhnikovia divaricata, Peucedanum japonicum, and Glehnia littoralis of the family Umbelliferae, have long been used as traditional herbal medicine in Asian countries. This study is the first to compare and analyse the relationship between antimicrobial activity, antioxidant activity, and total polyphenol content of Saposhnikovia divaricata, Peucedanum japonicum, and Glehnia littoralis. The four fractions of hydro-methanol extract of these 3 plants were evaluated using the disc diffusion method to determine the minimum inhibitory concentration against 5 different bacterial strains. The ethyl acetate fraction was the most effective against the bacterial strains investigated. The 1,1-diphenyl-2-picryl-hydrazyl free radical scavenging activity was evaluated to measure antioxidant activity. The antioxidant activity of the ether and ethyl acetate fractions was in the order of Saposhnikovia divaricata>Peucedanum japonicum>Glehnia littoralis. The total polyphenol content of Saposhnikovia divaricata was greater than that of Peucedanum japonicum and Glehnia littoralis. Even though the 3 plants belong to the same family and are used for similar medical purposes, their antimicrobial activity, antioxidant activity, and total polyphenol content was different. This study would help researchers to uncover the critical aspects of the activities possessed by plants.
As a traditional Chinese multiherbal formula, Yu-Ping-Feng (YPF) is frequently used to treat cold, flu and inflammation-associated diseases. We aimed to evaluate the immunostimulatory effects of polysaccharide isolated from YPF (YPF-PS) in vitro and in vivo. In in vitro experiment, macrophage cell proliferation, phagocytosis rate, cytokine and costimulatory molecule release, T lymphocyte proliferation, cell cycle distribution, CD4⁺ and CD8⁺ T cell percentages were determined. To investigate the in vivo effects of YPF-PS treatment, different doses YPF-PS were administered to chicken vaccinated against Newcastle disease. The immune organ index, lymphocyte proliferation, antibody titer, cell cycle distribution, and the cell percentage of CD4⁺ and CD8⁺ were assessed. In vitro results indicated that YPF-PS at 15.62 μg mL⁻¹ could increase the LPS-induced macrophage cell proliferation and phagocytosis rate significantly. The levels of cytokine (nitric oxide, tumor necrosis factor-alpha, interleukin-1 beta, interleukin-6, and interferon beta) and costimulatory molecules (CD80 and CD86) were also considerably enhanced. Moreover, YPF-PS could significantly enhance T lymphocyte proliferation individually or synergistically with phytohemagglutinin. It promoted lymphocyte entry into S and G2/M phases and increased the percentages of CD4⁺ and CD8⁺ T cells effectively. In addition, in vivo experiments showed that YPF-PS could enhance serum HI antibody titer. The results about T lymphocyte proliferation, cell cycle distribution, CD4⁺ and CD8⁺ cell percentages in chickens were also confirmed. YPF-PS has efficacious immunomodulatory properties and could be used as a new potential immune stimulator for food and medical purposes.
Introduction
4’-O-β-D-glucosyl-5-O-methylvisamminol (GML) is a conventional marker compound for quality control of Radix Saposhnikoviae. Despite that, neither pharmacodynamic or pharmacokinetic information is available with regard to GML. As such, the aim of thisstudy was to assess the conventional evaluation indices for the quality of Radix Saposhnikoviae.
Materials and methods
Pyretic animal model, hot plate test, and ear edema model were established to evaluate and compare the antipyretic, analgesic, and anti-inflammatory effect of the chromone derivativescimifugin, prime-O-glucosylcimifugin (PGCN), and GML in Radix Saposhnikoviae. High performance liquid chromatography separation and analysis was used to obtain pharmacokinetic parameters. Simulated gastric fluid and simulated intestinal fluid was used to investigate the metabolite profiles of PGCN and GML in gastrointestinal tract.
Results
Cimifugin exerted a marked dose-dependent antipyretic, analgesic, and anti-inflammatory effect, whereas the effects of PGCN were relatively lower. GML had feeble pharmacodynamic effects. Pharmacokinetic study showed that only cimifugin was detected in the plasma sample of cimifugin and PGCN-treated animals, with drug concentration in the former much higher than the latter. No components were traced in the plasma samples from GML-treated rats. Stability study showed that PGCN and GML was predominantly biotransformed into cimifugin and 5-O-methyvisammiol, respectively. The latter was proven to be extremely unstable in liver tissue homogenate and plasma.
Conclusions
A feeble antipyretic, analgesic, and anti-inflammatory activities was observed when GML was orally delivered. Given that Radix Saposhnikoviae extract is generally administered orally, we speculate that this compound might be a nonpharmacolagically active agent in real usage. Thus, it might be unscientific to evaluate the quality of Radix Saposhnikoviae based on the content of GML.
SUMMARY
GML-derived cimifugin, which represents the potential pharma codynamic component of Radix Saposhnikoviae chromones, in plasma was almost nil in contrast to cimifugin and PGCN. And thus, feeble antipyretic, analgesic, and anti-inflammatory activities were found with GML.
Abbreviations used: AUC:area under concentration-time curve, DNP:2,4-Dinitrophenol, HPLC:high performance liquid chromatography, HPLC-MS:high performance liquid chromatography- mass spectrography, GML:4’-O-β-D-glucosyl-5-O-methylvisamminol, MVL:5-O-methyvisammiol, PGCN:prime-O-glucosylcimifugin, SGF:alkaline phosphatase. SIF:simulated intestinal fluid
Ethnopharmacological relevance:
Chang-Kang-Fang formula (CKF), a multi-herb traditional Chinese medicinal formula, has been clinically used for treatment of irritable bowel syndrome (IBS). The mechanisms of CKF for treating IBS and the components that are responsible for the activities were still unknown.
Aim of the study:
To investigate the chemical profiles and effects of CKF on IBS model.
Materials and methods:
The chemical profiles of CKF were investigated by ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS/MS). On colon irritation induced rat neonates IBS model, the influence of CKF on neuropeptides, including substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and 5-hydroxytryptamine (5-HT), were measured by ELISA, and the effect on intestinal sensitivity was assessed based on the abdominal withdrawal reflex (AWR) scores. In addition, the activities of CKF against acetic acid-induced nociceptive responses and prostigmin methylsulfate triggered intestinal propulsion in mice were also evaluated.
Results:
80 components were identified or tentatively assigned from CKF, including 11 alkaloids, 20 flavanoids, 4 monoterpenoids, 9 iridoid glycoside, 9 phenylethanoid glycosides, 10 chromones, 7 organic acid, 3 coumarins, 2 triterpene and 5 other compounds. On IBS rat model, CKF was observed to reduce AWR scores and levels of SP, CGRP, VIP and 5-HT. Moreover, CKF reduced the acetic acid-induced writhing scores at all dosages and reduced the intestinal propulsion ration at dosage of 7.5 and 15.0 g/kg/d.
Conclusions:
CKF could alleviate the symptoms of IBS by modulating the brain-gut axis through increasing the production of neuropeptides such as CGRP, VIP, 5-HT and SP, releasing pain and reversing disorders of intestinal propulsion. Berberine, paeoniflorin, acteoside, flavonoids and chromones may be responsible for the multi-bioactivities of CKF.
Description of the drug: Long conical or long cylindrical, gradually tapering towards the lower part, some slightly tortuous, 15–30 cm long, 0.5–2 cm in diameter. Externally greyish-brown, rugged, with longitudinal wrinkles, numerous transverse-elongated lenticel-like protrudings and dotted raised rootlet scars. Root stock with obvious dense annulations, some annulations marked by brown hair-like remains of leaf bases. Texture light, easily broken, fracture uneven, bark brownish and cracked, wood yellowish. Odour, characteristic; taste, sweetish.
Vegetative but not reproductive stage of Saposhnikovia divaricate (Turxz.) schischk possesses pharmacological activities. However, our recent study showed that reproductive S. divaricate supplemented with polysaccharide showed evidently elevated pharmacological activities and increased cimifugin content in rat serum. The aims of present study were to assess the influence of polysaccharides on the chromones pharmacological activities in Radix Saposhnikoviae (RS), the dried root of vegetative stage of S. divaricate, and to explore the underlying mechanisms. Only cimifugin was detected in the plasma of chromone treated animals and RS polysaccharide significantly increased the plasma content of cimifugin. It was shown that neither cimifugin absorption nor glycoside components transformation in simulated digestive fluid was affected by RS polysaccharide. However, a significant promotion of transformation of cimifugin to more stable prime-O-glucosylcimifugin (PGCN) by RS polysaccharide, and a protective effect of polysaccharide on chromone components were observed in small intestine solutions. Meanwhile, RS polysaccharide produced a significant elevation of cimifugin and PGCN concentration in vivo. Based on these findings, we concluded that RS polysaccharide could greatly increase the content of cimifugin, which might be related to its degradation-proof effect on cimifugin, via transforming cimifugin to comparatively more stable PGCN and spatial structure protection.
Natural products have always been a pivotal source of new drug development. Dry roots of Saposhnikovia divaricata (Turcz.) Schischk. (Umbelliferae) is a perennial herb and is also known as Bang Pung in traditional medicine. Numerous in vitro and in vivo studies have revealed the diverse pharmacological effects of S. divaricata and its role in the treatment of various diseases. This herb has exhibited significant inhibitory effects against inflammation and associated disorders. The present study explored the ethnopharmacological applications and molecular mechanisms behind the anti-inflammatory effects of S. divaricata herb and a single compound blockade of multi-signaling inflammatory cascades. Taken together, this review provides insight into the potential role of S. divaricata against various inflammatory diseases.
Matrix metalloproteinases (MMPs) play an important role in normal physiological functions and pathological processes. They are involved in normal skin functions as well as in the aging, healing, embryonic development, reproduction, and inflammatory responses of skin. Previous studies report that both high MMP-1 and MMP-2 activities were found in the skin of patients with dermatitis, and large amounts of MMP-9 have been reported to be accumulated in unhealed wounds. Interleukin-8 (IL-8), a C-X-C chemokine, may mediate neutrophil recruitment and activation and is involved in various inflammatory skin diseases. In this study, eleven Chinese herbal medicines were analyzed for their potential as anti-inflammatory agents using human fibroblast WS-1 cell lines. The results indicate MMP-1 and -9, but not MMP-2, were induced by TNF-α treatment in WS-1 cells. However, when WS-1 cells were pre-treated with eleven Chinese herbal medicines before TNF-α stimulation, all these herbal medicines suppressed TNF-α-stimulated MMP-1 activity in WS-1 cells as analyzed by casein zymography. In addition, the suppression of MMP-9 activity was also observed when WS-1 cells were treated with either Paeonia suffruticosa, Scutellaria baicalensis, Saposhnikovia divaricata, Dioscorea opposita, Rubus chingii, or Salvia miltiorrhiza. Of which, R. chingii significantly inhibited IL-8 secretion induced by TNF-α treatment as well. These results revealed that some novel components present in these Chinese herbal medicines may be used for the treatment of inflammatory responses in skin cells.
In vitro Matrix metalloproteases-2 (MMP-2) inhibition assay and ultrafiltration liquid chromatography coupled to electrospray ionization tandem mass spectrometry (ultrafiltration LC-ESI-MSn) were combined to screen MMP-2 inhibitors from three Chinese medicinal herbs, Smilax glabra Roxb., Smilax china L. and Saposhnikovia divaricata (Turcz.) Schischk as potential functional food ingredients. Results showed that extracts of two Smilax species and the three major components resveratrol, engelitin and asibinn, and extract of S. divaricata and its major component 4′-O-β-D-glucosyl-5-O-methylvisamminol and three other chromones were strong MMP-2 inhibitors. Resveratrol and 4′-O-β-D-glucosyl-5-O-methylvisamminol had the highest binding affinity to MMP-2 and were the main contributors to the MMP-2 activity of Smilax and Saposhnikovia extracts, respectively. The present results further suggest that ultrafiltration LC-ESI-MSn is a valid, selective and effective method, and can be used to screen MMP-2 inhibitors from other plants such as grapes that contain resveratrol as functional foods and nutraceuticals.
A method employing high performance liquid chromatography-diode array detection (HPLC-DAD) has been developed to evaluate the quality of Saposhnikoviae Radix (SR) comprehensively through fingerprint analysis and multi-ingredient determination. In the fingerprint analysis, 12 out of 15 common peaks were identified with high performance liquid chromatography-tandem mass spectrometric (HPLC-MS/MS) and related references. To discriminate the 15 batches of samples collected from different origins, similarity analysis and principal component analysis (PCA) were performed. Meanwhile, the simultaneous quantification of four chromones was achieved by firstly applying a method of using a single standard to determine multiple components (SSDMC) with conversion factor. The method has the advantage of feasibility, economy and simplicity compared with traditional external standard method (ESM). The method possessed desirable linearity (R(2) ≥ 0.9997), precision (RSD <4.0%), accuracy (97.4 - 106.6%) and robustness. This study indicated that the method of fingerprint analysis integrated with multi-ingredient determination using the HPLC technique was reliable for the overall quality evaluation of SR.
A generalized method for analyzing the effects of multiple drugs and for determining summation, synergism and antagonism has been proposed. The derived, generalized equations are based on kinetic principles. The method is relatively simple and is not limited by 1) whether the dose-effect relationships are hyperbolic or sigmoidal, 2) whether the effects of the drugs are mutually exclusive or nonexclusive, 3) whether the ligand interactions are competitive, noncompetitive or uncompetitive, 4) whether the drugs are agonists or antagonists, or 5) the number of drugs involved.
To investigate the possible drug interaction with herbal medicine, furanocoumarin derivatives isolated from several Umbelliferous crude drugs were examined for their inhibitory effects on a typical human drug metabolizing enzyme, cytochrome P450 3A (CYP3A). Most furanocoumarins tested at 0.1 mM reduced microsomal testosterone 6beta-hydroxylation as an index of CYP3A activity to less than 50% of the control. In particular, the dimer and trimer derivatives of furanocoumarins showed striking inhibition, whose potencies were similar to that of a typical CYP3A inhibitor, ketoconazole. Preincubation of dimer types of furanocoumarins increased suppression but not most of the monomer derivatives, suggesting that the inhibition on CYP3A activity was caused by at least plural mechanisms. These results raised the possibility that the furanocoumarin containing herbal medicines may alter pharmacokinetics of co-ingested drugs similar to the case with grapefruit juice.
A series of polyacetylenes, falcarinone, panaxynol, falcarindiol, panaxydol, and panaxytriol, were isolated from Saposhnikovia divaricata (Turcz.) Schischk and Panax quinquefolium L. These polyacetylenes were identified as active principles on the inhibition of nitrite production by inducible nitric oxide synthase (iNOS). Treatment with 10 microM of panaxynol, falcarindiol, panaxydol and panaxytriol decreased the LPS/IFN-gamma-stimulated accumulation of nitrite by 71.92 +/- 3.07, 69.95 +/- 3.68, 45.48 +/- 6.11 and 36.85 +/- 8.80%, respectively. The IC50 value of falcarinone, panaxynol, falcarindiol, panaxydol and panaxytriol was > 20, 2.23, 1.98, 6.58 and 9.85 microM, respectively.
By activity-oriented separation using the writhing method in mice, the analgesic components of Saposhnikovia root (Saposhnikovia divaricata Schischkin; Umbelliferae) were identified to be chromones, coumarins, polyacetylenes and 1-acylglycerols. Two new components, divaricatol and (3'S)-hydroxydeltoin, were also isolated. The most potent analgesia was observed in chromones such as divaricatol, ledebouriellol and hamaudol, which inhibited writhing inhibition at an oral dose of 1 mg/kg in mice. Acylglycerols also showed inhibition significantly at a dose of 5 mg/kg. In some pharmacological tests using sec-O-glucosylhamaudol, the compound showed analgesia by the tail pressure and the Randall & Selitto methods, and its writhing inhibition was not reversed by naloxone.
To investigate the possible drug interaction with herbal medicine, hot water decoctions or 40% ethanol infusions of several Umbelliferous or Citrus crude drugs and their prescriptions were examined in vitro for their abilities to inhibit human cytochrome P450 3A (CYP3A). Addition of each decoction or infusion from Baizhi (Angelica dahurica and varieties), Qianghuo (Notopterygium incisum or N. forbesii), Duhuo (Angelica biserrata), Fangfeng (Saposhnikovia divaricata), Danggui (Angelicasinensis), Zhishi or Zhiqiao (Citrus aurantium) resulted in various degrees of human CYP3A inhibition as determined by microsomal testosterone 6beta-hydroxylation. The inhibitory potency was consistent with the abundance of the hydrophobic components for each sample. Experiments on the infusion of a Japanese Baizhi (BZ1) showed the major role of furanocoumarins on human CYP3A inhibition. Some of the crude drugs and a related prescription showed increased inhibition after the preincubation, suggesting the involvement of a mechanism-based inhibition. Some formulated prescriptions, however, showed intense inhibition with their hydrophobic fractions rather than with their hydrophobic fractions, suggesting that components other than furanocoumarins in herbal prescriptions may also cause CYP3A inhibition. These results indicate the necessity of intensive investigations on the possible drug interaction with traditional medicines.
In this study, six common tests for measuring antioxidant activity were evaluated by comparing four antioxidants and applying them to beverages (tea and juices): Trolox equivalent antioxidant capacity assay (TEAC I-III assay), Total radical-trapping antioxidant parameter assay (TRAP assay), 2,2-diphenyl-l-picrylhydrazyl assay (DPPH assay), N,N-dimethyl-p-phenylendiamine assay (DMPD assay), Photochemiluminescence assay (PCL assay) and Ferric reducing ability of plasma assay (FRAP assay). The antioxidants included gallic acid representing the group of polyphenols, uric acid as the main antioxidant in human plasma, ascorbic acid as a vitamin widely spread in fruits and Trolox as water soluble vitamin E analogue. The six methods presented can be divided into two groups depending on the oxidising reagent. Five methods use organic radical producers (TEAC I-III, TRAP, DPPH, DMPD, PCL) and one method works with metal ions for oxidation (FRAP). Another difference between these tests is the reaction procedure. Three assays use the delay in oxidation and determine the lag phase as parameter for the antioxidant activity (TEAC I, TRAP, PCL). They determine the delay of radical generation as well as the ability to scavenge the radical. In contrast, the assays TEAC II and III, DPPH, DMPD and FRAP analyse the ability to reduce the radical cation (TEAC II and III, DPPH, DMPD) or the ferric ion (FRAP). The three tests acting by radical reduction use preformed radicals and determine the decrease in absorbance while the FRAP assay measures the formed ferrous ions by increased absorbance. Gallic acid was the strongest antioxidant in all tests with exception of the DMPD assay. In contrast, uric acid and ascorbic acid showed low activity in some assays. Most of the assays determine the antioxidant activity in the micromolar range needing minutes to hours. Only one assay (PCL) is able to analyse the antioxidant activity in the nanomolar range. Black currant juice showed highest antioxidant activity in all tests compared to tea, apple juice and tomato juice. Despite these differences, results of these in vitro assays give an idea of the protective efficacy of secondary plant products. It is strongly recommended to use at least two methods due to the differences between the test systems investigated.
Cimifugin, one of the principal analgesic and anti-hypertensive component of Fang-Feng, was isolated from the root of Saposhnikovia divaricata. Reversed-phase high-performance liquid chromatography was employed to determine the contents of cimifugin in Fang-Feng. The separation was performed on a LiChrospher 100 RP-18e column (4 x 250 mm) by isocratic elution with acetonitrile-water (18:82) as the mobile phase at a flow-rate of 1.0 mL/min, the temperature being kept at 40°C and the detection set at 220 nm. Methylparaben was used as the internal standard. The regression equation revealed linear relationship between the peak-area ratio (marker substance/internal standard) and concentration. We found that Fang-Feng extracted with twenty-fold of 70% aqueous methanol under sonication for 40 minutes gives the best yield of cimifugin. The contents of cimifugin in commercially available Saposhnikoviae Radices have been determined and the amount was found to vary from 2.66 x 10 -2% to 0.19 x 10 -2%. This HPLC method can rapidly and easily quantify the cimifugin present in Fang-Feng and can be used for quality control of this commercial crude drug.
A method for the screening of antioxidant activity is reported as a decolorization assay applicable to both lipophilic and hydrophilic antioxidants, including flavonoids, hydroxycinnamates, carotenoids, and plasma antioxidants. The pre-formed radical monocation of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS*+) is generated by oxidation of ABTS with potassium persulfate and is reduced in the presence of such hydrogen-donating antioxidants. The influences of both the concentration of antioxidant and duration of reaction on the inhibition of the radical cation absorption are taken into account when determining the antioxidant activity. This assay clearly improves the original TEAC assay (the ferryl myoglobin/ABTS assay) for the determination of antioxidant activity in a number of ways. First, the chemistry involves the direct generation of the ABTS radical monocation with no involvement of an intermediary radical. Second, it is a decolorization assay; thus the radical cation is pre-formed prior to addition of antioxidant test systems, rather than the generation of the radical taking place continually in the presence of the antioxidant. Hence the results obtained with the improved system may not always be directly comparable with those obtained using the original TEAC assay. Third, it is applicable to both aqueous and lipophilic systems.
Coumarin and its 4-OH and 7-OH derivatives, as well as o-, m- and p-coumaric acid were tested against P-815 and P-388 tumor cells in vitro. In addition, the compounds were investigated in various in vitro immunological test systems and genuine coumarin was tested furthermore against the Sarcoma-180 in CD1 mice. In vivo, coumarin showed only a moderate antitumor effect against the allogeneic Sarcoma-180 at concentrations of 10 and 40 mg/kg, with inhibition rates of 49 and 60%, respectively. However, both concentrations were markedly toxic. In vitro all compounds were more or less cytotoxic against P-815 and P-388 tumor cell lines starting at a concentration of 100 micrograms/ml. At subtoxic concentrations (less than or equal to 10 micrograms/ml) the samples showed no mitogenic activity against murine spleen lymphocytes and PHA costimulated human peripheral blood lymphocytes. Furthermore, with the coumarin derivatives neither cytotoxic macrophages could be induced against P-815 tumor cells nor an increased release of Il-2 and TNF-alpha could be observed. Only 7-OH coumarin, in concentrations of 2 and 20 micrograms/ml, caused a strong increase in phagocytosis of 124 and 84% in both, human peripheral blood granulocytes and murine peritoneal macrophages, respectively.
An acidic polysaccharide, named saposhnikovan C, was isolated from the roots and rhizomes of Saposhnikovia divaricata Schischk. It was homogeneous as judged by electrophoresis and gel chromatography, and showed remarkable reticuloendothelial system-potentiating activity in a carbon clearance test. It is composed of D-galacturonic acid:L-rhamnose:L-arabinose:D-galactose in a molar ratio of 27:7:8:8, and its molecular mass was estimated to be 132000. About 30% of the D-galacturonic acid residues exist as the methyl esters. O-Acetyl groups were identified, and the content amounted to 3.3%. Methylation analysis, carbon-13 nuclear magnetic resonance, and controlled Smith degradation studies indicated the structural features. It has a pectin-like rhamnogalacturonan backbone with branched arabinan and galactan side chains.
The effects of various coumarins (i.e. esculetin, daphnetin and fraxetin) on the formation of the 5-lipoxygenase product, 5-HETE, and the cyclooxygenase product, HHT, were studied. Esculetin (6,7-dihydroxycoumarin) was found to inhibit the formation of 5-HETE more strongly than HHT; its concentrations for 50% inhibition (IC50) were 1.46 +/- 1.02 microM for the formation 5-HETE and 57.3 +/- 17.3 microM for the formation of HHT. Daphnetin (7,8-dihydroxycoumarin) and fraxetin (6-methoxy-7,8-dihydroxycoumarin) also inhibited the formation of the 5-lipoxygenase product, 5-HETE, and the cyclooxygenase product, HHT; their IC50 values were, respectively, 6.90 +/- 2.07 microM and 2.57 +/- 0.088 microM for the formation of 5-HETE and 139.0 +/- 30.0 microM and 532.5 +/- 33.0 microM for the formation of HHT. The monohydroxy coumarin derivatives umbelliferone (7-hydroxycoumarin) and scopoletin (6-methoxy-7-hydroxycoumarin) and the coumarin glucosides fraxin (6-methoxy-7,8-dihydroxycoumarin 8-O-D-glucoside) and esculin (6,7-dihydroxycoumarin 6-O-D-glucoside) also inhibited the formation of 5-HETE, though less strongly. 4-Hydroxycoumarin and coumarin had no effect on either 5-lipoxygenase or cyclooxygenase at concentrations of up to 1 mM. Esculetin inhibited the formation of 5-HETE noncompetitively. In contrast, the dimethoxycoumarin fraxidin (6,8-dimethoxy-7-hydroxycoumarin) inhibited the formation of HHT more strongly than the formation of 5-HETE at a concentration of 1 mM.
Terminal differentiation of human promyelocytic leukemia (HL-60) cells can be induced by a variety of chemical agents and this process can be monitored readily by the generation of morphologically, histochemically, and functionally mature granulocytes and monocytes/macrophages. The availability of this model has heightened interest in the possible therapeutic role of inducers of myeloid differentiation for the treatment of leukemia and other neoplasms. In addition, however, potent cancer chemopreventive agents induce HL-60 cell differentiation at very low dose levels. Thus, as part of our search for natural product chemopreventive agents, extracts derived from nearly 400 plants were tested for their potential to induce HL-60 cell differentiation. As a result, 17 plant extracts were judged to be active (ED50 values < or = 4 micrograms/ml). One of most potent leads was an extract derived from Dirca occidentalis Gray (Thymelaeaceae) (ED50, 0.14 micrograms/ml), and bioassay-guided fractionation led to the identification of genkwanin (I), (+/-)-lariciresinol (II) and sitoindoside II (IV) as active principles, with ED50 values of 18.3, 1.1 and 0.069 microM, respectively. Based on these data, we conclude that the HL-60 cell differentiation system is a valid and useful model for the discovery of natural product cancer chemopreventive or chemotherapeutic agents.
The beneficial effects of moderate red wine consumption have been attributed, in part, to the presence of antioxidant components. Oxidant stress is an activating stimulus for the NF (nuclear factor)-KB/Rel family of transcription factors, which have binding sites in the promoter regions of many genes involved in inflammatory and immune responses. The effect of lipopolysaccharide (LPS)-stimulated activation of NF-KB and the subsequent production of tumor necrosis factor alpha (TNF-alpha) and NO was determined in the macrophage cell line RAW 264.7. Unexpectedly, the wine polyphenolics quercetin and resveratrol and the antioxidant N-acetylcysteine (NAC) did not inhibit LPS-induced activation of the NF-KB complex p50/65, as determined by mobility shift. Quercetin inhibited LPS-induced p50/50. Northern blot analysis indicated that quercetin (0.1 and 0.2 mM) inhibited LPS-dependent production of inducible nitric oxide synthase (iNOS) mRNA and decreased NO release, as measured by the Griess reaction. This flavonoid had no effect on LPS-induced TNF-alpha mRNA, but decreased LPS-stimulated TNF-alpha release, as measured by ELISA. Resveratrol (0.05 and 0.1 mM) posttranscriptionally decreased LPS-induced nitrite release. It increased basal levels of TNF-alpha mRNA and protein and enhanced LPS-induced TNF-alpha mRNA and cytokine release. Our results do not support the view that wine antioxidants inhibit LPS-induced NF-KB activation but instead that they have a more selective action on genes activated by LPS.
Twenty-seven Citrus flavonoids were examined for their activity of induction of terminal differentiation of human promyelocytic leukemia cells (HL-60) by nitro blue tetrazolium (NBT) reducing, nonspecific esterase, specific esterase, and phagocytic activities. 10 flavonoids were judged to be active (percentage of NBT reducing cells was more than 40% at a concentration of 40 microM), and the rank order of potency was natsudaidain, luteolin, tangeretin, quercetin, apigenin, 3, 3, '4, '5, 6, 7, 8-heptamethoxyflavone, nobiletin, acacetin, eriodictyol, and taxifolin. These flavonoids exerted their activity in a dose-dependent manner. HL-60 cells treated with these flavonoids differentiated into mature monocyte/macrophage. The structure-activity relationship established from comparison between flavones and flavanones revealed that ortho-catechol moiety in ring B and C2-C3 double bond had an important role for induction of differentiation of HL-60. In polymethoxylated flavones, hydroxyl group at C3 and methoxyl group at C8 enhanced the differentiation-inducing activity.
L-Arginine derived nitric oxide (NO) and its derivatives, such as nitrogen dioxide and peroxynitrite, play a role in inflammation and also possibly in the multistage process of carcinogenesis. Four furanocoumarins and eight chromones isolated from the dried root of Saposhnikovia divaricata (Fang Feng in Chinese) and evaluated for their effects on the synthesis of NO induced by lipopolysaccharide (LPS) in macrophage cell line RAW 264.7. The inhibition of nitrite production, as an index for NO released from the macrophage cells, was quantitatively analyzed by Griess reaction. The results showed that imperatorin and deltoin are potential NO production inhibitor, and their IC50 values for inhibition of nitrite production were 17.3 and 11.6 microg/ml, respectively. Western-blot analysis demonstrated that iNOS enzyme activity was not inhibited by treatment with imperatorin or deltoin, but revealed that both compounds inhibited the expression of the iNOS protein.
Matrix degradation products such as fragmented hyaluronan (HA) display important proinflammatory effects on renal tubular epithelial cells (TECs) and macrophages (MPhis). We hypothesized that HA could up-regulate cyclooxygenase type 2 (COX-2) in these cells and that the subsequent production of thromboxane A2 (TXA2) could play a role in inflammatory renal lesions.
We used an in vitro approach to examine the expression of COX-1 and COX-2 and the production of TXA2 in response to fragments of HA. COX-2 mRNA, protein, and the resulting TXA2 production were measured in CD44-positive, HA-responsive cells lines of TECs and MPhi. COX-2 mRNA was also measured in vivo in MRL-Faslpr mice and in mice with anti-glomerular basement membrane (anti-GBM) nephritis.
In TECs and MPhis, HA increased the steady-state COX-2 mRNA and protein levels markedly, whereas COX-1 mRNA levels did not change. The HA-induced response was comparable to lipopolysaccharide stimulation. In comparison with MPhi, the response was much weaker in TECs. Likewise, the production of TXA2 in response to HA was markedly increased in MPhi, but less in TECs. In TECs and in MPhi, the HA-stimulated TXA2 synthesis was inhibited with the COX-2-selective inhibitors SC58125 (12.5 micromol/L) or celecoxib (0.25 to 5.00 micromol/L). COX-2 mRNA levels were increased in nephritic mice with MRL-Faslpr lupus nephritis and in mice with anti-GBM disease.
HA is a proinflammatory factor that stimulates COX-2 expression and subsequent TXA2 production. Since HA accumulates markedly in renal injury, we speculate that this matrix molecule could therefore play a significant role in thromboxane-mediated immune events in the kidney.
Two drugs that produce overtly similar effects will sometimes produce exaggerated or diminished effects when used concurrently. A quantitative assessment is necessary to distinguish these cases from simply additive action. This distinction is based on the classic pharmacologic definition of additivity that, briefly stated, means that each constituent contributes to the effect in accord with its own potency. Accordingly, the relative potency of the agents, not necessarily constant at all effect levels, allows a calculation using dose pairs to determine the equivalent of either agent and the effect by using the equivalent in the dose-response relation of the reference compound. The calculation is aided by a popular graph (isobologram) that provides a visual assessment of the interaction but also requires independent statistical analysis. The latter can be accomplished from calculations that use the total dose in a fixed-ratio combination along with the calculated additive total dose for the same effect. Different methods may be used, and each is applicable to experiments in which a single drug is given at two different sites. When departures from additivity are found, whether in "two-drug" or "two-site" experiments, the information is useful in designing new experiments for illuminating mechanisms. Several examples, mainly from analgesic drug studies, illustrate this application. Even when a single drug (or site) is used, its introduction places it in potential contact with a myriad of chemicals already in the system, a fact that underscores the importance of this topic in other areas of biological investigation.
Carnosol is a naturally occurring phytopolyphenol found in rosemary. Carnosol functions as antioxidant and anticarcinogen. In the present study, we compared the antioxidant activity of carnosol and other compounds extracted from rosemary. Carnosol showed potent antioxidative activity in alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) free radicals scavenge and DNA protection from Fenton reaction. High concentrations of nitric oxide (NO) are produced by inducible NO synthase (iNOS) in inflammation and multiple stages of carcinogenesis. Treatment of mouse macrophage RAW 264.7 cell line with carnosol markly reduced lipopolysaccharide (LPS)-stimulated NO production in a concentration-related manner with an IC50 of 9.4 microM; but other tested compounds had slight effects. Western blot, reverse transcription-polymerase chain reaction, and northern blot analyses demonstrated that carnosol decreased LPS-induced iNOS mRNA and protein expression. Carnosol treatment showed reduction of nuclear factor-kappaB (NF-kappaB) subunits translocation and NF-kappaB DNA binding activity in activated macrophages. Carnosol also showed inhibition of iNOS and NF-kappaB promoter activity in transient transfection assay. These activities were referred to down-regulation of inhibitor kappaB (IkappaB) kinase (IKK) activity by carnosol (5 microM), thus inhibited LPS-induced phosphorylation as well as degradation of IkappaBalpha. Carnosol also inhibited LPS-induced p38 and p44/42 mitogen-activated protein kinase (MAPK) activation at a higher concentration (20 microM). These results suggest that carnosol suppresses the NO production and iNOS gene expression by inhibiting NF-kappaB activation, and provide possible mechanisms for its anti-inflammatory and chemopreventive action.
In the present study, we tested ethanolic extracts from 10 Chinese herbs for their effects on K562, Raji, Wish, HeLa, Calu-1, and Vero tumor cells proliferation. On a percentage basis, panaxynol purified from Saposhnikovae divaricata had the highest inhibitory activity on various tumor cells proliferation. Cell-cycle analysis indicated that panaxynol arrested the cell cycle progression of tumor cells from the G1 transition to the S phase. In an attempt to further localize the point in the cell cycle where arrest occurred, gene expression of cyclin E, a key regulatory event leading to the G1/S boundary was examined. Results indicated that the levels of cyclin E mRNA in various tumor cells were decreased by panaxynol. Thus, the suppressant effects of panaxynol on proliferation of various tumor cells appeared to be mediated, at least in part, through impairments of cyclin E mRNA levels and arresting cell cycle progression in the cells.
Pretreatment of mice with lyophilized hot water extracts of five poly-herbal formula protected them from lethal infection by E. coli. ESR spectroscopy shows that these extracts produced radicals under alkaline condition, and scavenged radicals such as superoxide anion, hydroxyl radical and nitric oxide (NO) radical. There was a positive relationship between their radical intensity and radical scavenging activity. Among the extracts, HD-02 efficiently inhibited the production of NO and citrulline, and the expression of inducible NO synthase (iNOS) mRNA by LPS-stimulated mouse macrophage-like cells Raw 264.7. DLH-3073 not only inhibited the LPS-stimulated NO production at lower concentration, but also induced NO production at higher concentrations, suggesting the presence of two different antagonizing components in the DLH-3073 extract. These data suggest that poly-herbal extracts may alleviate radical-mediated diseases.
Sutherlandia frutescens is a South African herb used traditionally by the natives to treat cancer, and more recently to improve the overall health in HIV/AIDS patients. Gas chromatography/mass spectrometer profiling and liquid chromatographic/mass spectral investigation confirmed and quantified the presence of canavanine, GABA and arginine in the herbal preparation used in this study. In vitro study demonstrated a concentration dependent effect of Sutherlandia on several tumor cell lines, with 50% inhibition (IC50) of proliferation of MCF7, MDA-MB-468, Jurkat and HL60 cells at 1/250, 1/200, 1/150 and 1/200 dilutions, respectively. Sutherlandia treatment did not induce HL60 differentiation along the macrophage/monocyte or granulocyte lineage. It demonstrated antioxidant activity in reducing free radical cations with an estimated activity of 0.5 microl of Sutherlandia extract equivalent to that of 10 microM of Trolox. However, it did not significantly suppress lipopolysaccharide stimulated nitric oxide production by murine macrophage/monocyte RAW 264.7 cells, nor did it significantly inhibit IL-1beta and TNF-alpha mRNA expression in RAW 264.7 cells. In conclusion, Sutherlandia ethanolic extract showed a concentration dependent antiproliferative effect on several human tumor cell lines but did not show significant antioxidant effects. Further studies are needed to explore the activities of this multipurpose South African herbal preparation.
The effects of falcarindiol on the expression of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide/interferon-gamma (LPS/IFN-gamma) in rat primary astrocytes were investigated. The molecular mechanisms underlying falcarindiol that confers its effect on iNOS expression were also elucidated. Falcarindiol abrogated the LPS/IFN-gamma-mediated induction of iNOS by about 80%. Falcarindiol attenuated the induction of iNOS in a concentration-dependent manner. The inhibitory effect of falcarindiol on iNOS induction was attributable to decrease in the protein content and the mRNA level of iNOS. Treatment with 50 microM of falcarindiol for 30 min decreased LPS/IFN-gamma-induced nuclear factor-kappaB (NF-kappaB) activation by 32%. Treatment with 50 microM of falcarindiol for 60 min diminished the LPS/IFN-gamma-mediated activation of IkappaB kinase-alpha (IKK-alpha) and IKK-beta by 28.2 and 29.7%, respectively. Falcarindiol modulated the nuclear translocation of signal transducer and activator of transcription 1 (Stat1) in a time-dependent manner. Falcarindiol (50 microM) decreased the tyrosine phosphorylation of janus kinase 1 (JAK1) by 84.8% at 5 min. Falcarindiol also abrogated the tyrosine phoshorylation of JAK2 by 82.3% at 10 min.The present study demonstrates that falcarindiol attenuated the activation of IKK and JAK contributing to the blockade of activation of NF-kappaB and Stat1, thereby leading to the suppression of iNOS expression.
One-step RT-PCR to detect cytokine/chemokine induction in macrophages
- M Bongers
- Leihl E Barsig
Bongers M, Leihl E and Barsig J: One-step RT-PCR to detect
cytokine/chemokine induction in macrophages. Focus 21: 61-68,
1999.
Effect of citrus flavinoids on HL-60 cell differentiation
- S Kawaii
- Y Tomono
- E Katase
- K Ogawa
- M Yano
Kawaii S, Tomono Y, Katase E, Ogawa K and Yano M: Effect
of citrus flavinoids on HL-60 cell differentiation. Anticancer
Res 19: 1261-1270, 1999.