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ABSTRACT: The root of Astragalus membranaceus (AR), which has been widely used in Traditional Chinese herbal formulae for treating foot ulcer, was found to exhibit anti-inflammatory property, but its molecular mechanism still remains unknown. We previously identified the anti-inflammatory sub-fraction using bioassay-guided fractionation. The objective of the present study was to investigate the anti-inflammatory mechanism of the major active fraction (MAF) (0.039 to 0.156 mg/mL) using lipopolysaccharide (LPS)-stimulated mouse macrophage RAW 264.7 cells. MAF was shown to inhibit LPS-induced mRNA and protein expression of inducible nitric oxide synthase by 54.7% and 65.1%, respectively. Additionally, MAF down-regulated the protein expression of cyclooxygenase-2 and MAPK regulator by 45.0% to 74.6%, as well as the reduction of DNA binding activity of nuclear factor kappa B (NFκB) by 66.5%. It also attenuated the production of prostaglandin E2 , interleukin-1 beta (IL-1β), IL-6 and tumor necrosis factor alpha by 21.2% to 86.2%. Furthermore, the chemical constituents of MAF were identified. A total of 13 known chemical compounds were found in MAF, including five isoflavonoids and eight saponins. In conclusion, a bioactive fraction of AR was identified which possessed anti-inflammatory property by reducing the release of inflammatory mediators and inactivation of NFκB through MAPK signalling pathway. Copyright © 2013 John Wiley & Sons, Ltd.
Phytotherapy Research 05/2013; · 2.09 Impact Factor
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ABSTRACT: ETHNOPHARMACOLOGICAL RELEVANCE: The herbal formulaDG, containing roots of Salvia miltiorrhiza (Danshen) and Pueraria lobata (Gegen),has long history in treatingcardiovascular diseases. It has been shown to be able to reduce intima-media thickening in coronary patients in our previous clinical study.Since intima-media thickening is the hallmark of atherosclerotic disease,the etiology of which is inflammation of arterial wall, the mechanism underlying the effect of DGmay be related to its anti-inflammatory activities. AIM OF STUDY: The present study aims to determine the anti-inflammatory activity of DG and elucidate its underlying mechanisms with regards to its molecular basis of action. MATERIAL AND METHOD: The anti-inflammatory effect of DG was studied by using lipopolysaccharide (LPS)-stimulated activation of nuclear factor κB (NFκB) pathway and subsequent productions of inflammatory mediators, including nitric oxide (NO), prostaglandin E(2) (PGE(2)), interleukin-1 beta (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α) and macrophage chemotactic protein-1 (MCP-1), in mouse RAW 264.7 macrophages. RESULTS: The present study demonstrated that DG could suppress productions of NO and PGE(2)through inhibition of iNOS and COX-2 genes.Also, DG could inhibitproductions of IL-1β, IL-6 and MCP-1, but not TNF-α, through inhibition of respectivemRNA expressions. Further investigations showedthe inhibitory effect of DG on activation of IKKα/β and degradation of IκBα, thuspreventing nuclear translocation of NFκB. All these results suggested the inhibitory effects of DG on productions of inflammatory mediators through inhibition of the NFκB pathway. CONCLUSIONS: The inhibitory effects of DG on production of inflammatory mediators by LPS-stimulated RAW 264.7 macrophages, are accomplished by inhibitingthe nuclear translocation of NFκB through inactivatingIKKα/β and preventing degradation ofIκBα.
Journal of ethnopharmacology 12/2012; · 2.32 Impact Factor
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ABSTRACT: Herba Epimedii, an herb commonly used in East Asian medicine, is commonly used for treatment of impotence, osteoporosis and many inflammatory conditions in traditional Chinese medicine. Recent studies revealed that Herba Epimedii also has anti-tumor or anti-cancer activities, which may possibly be mediated through anti-angiogenesis. This study aims to examine and confirm the anti-angiogenic activity in the herb using both in vivo and in vitro approaches. The 95% ethanol extract and four subsequent fractions (n-hexane, ethyl acetate (EA), n-butanol and aqueous fractions) of Herba Epimedii were tested on the zebrafish model by the quantitative assay for endogenous alkaline phosphatase; then, the active fraction was further tested on Tg(fli1a:EGFP)y1 zebrafish embryos and human umbilical vein endothelial cells (HUVECs) for the anti-angiogenic effects. In addition, the action mechanism of Herba Epimedii was further investigated on wild-type zebrafish embryos and HUVECs. The EA fraction showed anti-angiogenic effects in both in vivo and in vitro models. Further experiments demonstrated that it might affect angiogenesis by acting on multiple molecular targets in zebrafish embryos and ERK signaling pathway in HUVECs. In conclusion, Herba Epimedii can inhibit angiogenesis, which may be the mechanism for its anti-inflammatory, anti-tumor and anti-cancer actions. Copyright © 2012 John Wiley & Sons, Ltd.
Phytotherapy Research 11/2012; · 2.09 Impact Factor
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ABSTRACT: Kaempferia galanga L. (Zingiberaceae) is an aromatic herb and a popular spice used as condiment in Asian cuisine. The ethanol extract of the dried plant and its successive four sub-fractions were investigated on zebrafish model by quantitative endogenous alkaline phosphatase assay. Both n-hexane and ethyl acetate fractions had antiangiogenic activity, and two major active components (trans-ethyl p-methoxycinnamate and kaempferol) showed potent antiangiogenic effects on wild-type zebrafish. Due to its much stronger effect and no antiangiogenic activity reported, trans-ethyl p-methoxycinnamate was further investigated for its action mechanism. It dose-dependently inhibited vessel formation on both wild- and Tg(fli1a:EGFP)y1-type zebrafish embryos. The semi-quantitative reverse transcription polymerase chain reaction assay suggested trans-ethyl p-methoxycinnamate affect multiple molecular targets related to angiogenesis. In vitro, it specifically inhibited the migration and tube formation of human umbilical vein endothelial cells. In vivo, it could block bFGF-induced vessel formation on Matrigel plug assay.
Journal of Agricultural and Food Chemistry 10/2012; · 2.82 Impact Factor
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Qi Zhang,
Chi Chun Fong,
Wai Kin Yu,
Yao Chen,
Fan Wei,
Chi Man Koon,
Kit Man Lau,
Ping Chung Leung, Clara Bik San Lau,
Kwok Pui Fung,
Mengsu Yang
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ABSTRACT: Astragali Radix (AR) and Rehmanniae Radix (RR) have long been used in traditional Chinese Medicine and as the principal herbs in treating diabetic foot ulcer. In this study, we investigated the effect of NF3, which comprises of AR and RR in the ratio of 2:1(w/w), on skin fibroblast cell migration and the activation of selected genes and proteins related to wound healing. Human skin fibroblast cell line Hs27 was treated with NF3 at 4mg/ml for 24h, and in vitro scratch wound healing and quantitative cell migration assays were performed, respectively. The expression of transformation growth factor (TGF-β1) and bone morphogenetic protein 6 (BMP6) in Hs27 cells with or without NF3 treatment was analyzed by western blot analysis. In addition, the expression of a panel of genes involved in human TGF-β signaling pathway was analyzed in Hs27 cells upon NF3 treatment (4mg/ml, 24h) by quantitative real-time PCR (qRT-PCR). Furthermore, the expression of several genes and proteins associated with ECM synthesis was investigated by qRT-PCR analysis or/and ELISA techniques. The results suggested that NF3 promoted the migration of human skin fibroblast cells. Western blot analysis demonstrated that NF3 up-regulated TGF-β1 and BMP-6 synthesis. qRT-PCR analysis revealed that the expression of 26 genes in Hs27 cells was changed upon NF3 induction, including TGF-β superfamily ligands and down stream effectors genes, and genes involved in TGF/Smad pathway, and Ras/MAPK (non-Smad) pathway. Among the extracellular matrix (ECM)-related molecules, it was found that NF3 up-regulated the expression of type I and III collagens, fibronectin as well as TIMP-1, and down-regulated the MMP-9 expression in skin fibroblast cells. This study demonstrated that herb formula NF3 could enhance skin fibroblast cell migration and activated genes involved in TGF-β1 pathway. NF3 could regulate gene transcription for extracellular matrix synthesis via the Smad pathway, and gene transcription for cell motility via the Ras/MAPK (non-Smad) pathway.
Phytomedicine: international journal of phytotherapy and phytopharmacology 10/2012; · 2.17 Impact Factor
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ABSTRACT: The root of Rehmannia glutinosa (RR) is commonly used to reduce inflammation in various traditional Chinese herbal formulae; however, little is known regarding its active component(s). Aim of study: The objective of the present study was to examine the active component(s) responsible for the anti-inflammatory activity of RR via anti-nitric oxide production assay-guided fractionation; and the underlying anti-inflammatory mechanism of action of such component(s) was further investigated.
Anti-nitric oxide (NO) activities with lipopolysaccharides (LPS)-stimulated RAW264.7 murine macrophages was used as screening platform. Gene, protein and inflammatory mediators' expression were also studied using real-time PCR, western blotting and ELISA, respectively.
Using anti-NO assay-guided fractionation, sub-fraction C3 (from 31.25 to 62.5μg/ml, p=0.001 to 0.01) possessed 100-fold more potent anti-inflammatory effect than that of the aqueous extract of RR. Characterization of C3 showed that the anti-inflammatory effect could be partly due to the presence of rehmapicrogenin, which could significantly inhibit NO production (p<0.001). C3 was further demonstrated in blocking inflammation by inhibiting gene (p<0.001) and protein expression of inducible NO synthase (iNOS) dose-dependently. Besides, C3 also significantly inhibited the production of prostaglandin E(2) (p<0.001 to 0.01), IL-6 (p<0.001 to 0.05) and COX-2 (p<0.05).
Rehmapicrogenin was, for the first time, shown to possess nitric oxide inhibitory activities. Bioassay-guided fractionation demonstrated that rehmapicrogenin-containing subfraction C3 exhibited potent anti-inflammatory effect by inhibiting iNOS, COX-2 and IL-6, while rehmapicrogenin was only partially responsible for the anti-inflammatory effect of RR.
Journal of ethnopharmacology 10/2012; 143(3):867-75. · 2.32 Impact Factor
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ABSTRACT: Erxian Decoction (EXD), a traditional Chinese herbal formula, has been used to treat menopausal symptoms and other aging diseases for several decades. Recently, our laboratory found that EXD could inhibit the proliferation of breast cancer cells. This activity may be mediated by anti-angiogenic action. To investigate the anti-angiogenic activity of EXD, its inhibitory effect on blood vessel formation was evaluated using both wild type and transgenic zebrafish embryos with fluorescent vasculature in vivo. Both semi-quantitative and real-time qPCR were carried out to evaluate the effect of EXD on the expression of several genes closely associated with angiogenesis in zebrafish. EXD was found to inhibit vessel formation in zebrafish embryos in a dose- and time-dependent manner. Furthermore, it reduced the mRNA expression of vascular endothelial growth factor A (VEGF-A) and the protein level of hypoxia inducible factor 1α (HIF-1α) in the embryos, suggesting the involvement of HIF-1 mediated VEGF-A signaling pathway in the anti-angiogenic action of EXD. The anti-angiogenic activity of EXD provides new insights to its clinical application and may in the future lead to the development of potential drugs for treating various cancers, especially in menopausal period.
Biological & Pharmaceutical Bulletin 09/2012; · 1.66 Impact Factor
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ABSTRACT: The roots of Salvia miltiorrhiza (Danshen) and Pueraria lobata (Gegen) are principle herbs of Chinese herbal formulae which have long been used to treat cardiovascular diseases.
The present study validated the anti-atherogenic effects of three extracts, Danshen alone (DE), Gegen alone (GE) as well as DGE and interpreted their combination effects statistically.
The anti-atherogenic effects of the three extracts were studied in three assays with regards to inflammation, foam cell formation and vascular smooth muscle cell (vSMC) proliferation using lipopolysaccharide (LPS)-induced nitric oxide production model, macrophage foam cell formation model and platelet-derived growth factor (PDGF)-induced vSMC proliferation model, respectively. The combination effects of DGE were statistically analyzed using combination index (CI) and fixed-ratio experimental design.
The anti-atherogenic effects of the three extracts including anti-inflammation, anti-foam cell formation and anti-vSMC proliferation were demonstrated in this study. Their combination effects in anti-inflammation, anti-foam cell formation and anti-vSMC proliferation were found to be synergistic, additive and antagonistic, respectively.
This study provided scientific support for the combination use of DGE on atherosclerosis and presented one of the first applications of statistical interpretations of the combination effects of the 2-herb formula.
Journal of ethnopharmacology 08/2012; 143(3):859-66. · 2.32 Impact Factor
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ABSTRACT: Foot ulceration, if not treated properly, will eventually result in amputation. Inflammation may impede the wound healing process if not properly controlled. The root of Astragalus membranaceus (AR) is one of the Chinese herbs commonly found in Chinese herbal formulae used for treating foot ulcer. In this study, we aimed to identify the active fractions and/or compounds from AR aqueous extract, which are responsible for the anti-inflammatory effect using in vitro bioassay-guided fractionation. The anti-inflammatory effect was monitored by the inhibition of nitric oxide (NO) released from lipopolysaccharide-stimulated mouse macrophage RAW 264.7 cells after treated with AR aqueous extract or its fractions and isolated components. Two major active fractions (P2-3-2-2-2 and P2-3-2-2-3) were found to significantly inhibit NO production at 0.156 mg/mL (p < 0.01). In addition, three chemical components (formononetin, calycosin and astragaloside IV) were successfully isolated from P2-3-2-2-3. Only formononetin could significantly inhibit NO production (p < 0.01), whereas the other two components had no significant effects at concentrations ranging from 0.039 to 0.156 mg/mL. In conclusion, two major anti-inflammatory active fractions that may enhance wound healing were identified, and formononetin was one of the active ingredients in the active fractions. Copyright © 2012 John Wiley & Sons, Ltd.
Phytotherapy Research 06/2012; · 2.09 Impact Factor
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Wing-Sum Siu,
Hing-Lok Wong,
Ching-Po Lau,
Wai-Ting Shum,
Chun-Wai Wong,
Si Gao,
Kwok-Pui Fung, Clara Bik-San Lau,
Leung-Kim Hung,
Chun-Hay Ko,
Ping-Chung Leung
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ABSTRACT: An innovative anti-osteoporosis herbal formula containing Epimedii Herba, Ligustri Lucidi Fructus and Psoraleae Fructus (ELP) has been previously shown its bone protecting effects in ovariectomized osteoporotic rats and also in post-menopausal osteopenic women. This study aimed to investigate the efficacy of ELP against bone loss during physical inactivity or weightlessness. A hindlimb unloading tail-suspended rat model was used for studying the effects of ELP on bone mineral density (BMD) and bone micro-architecture. For in vitro mechanistic studies, rat mesenchymal stem cells (MSCs) and mouse macrophage cells (RAW264.7) were used for studying the effects of ELP on osteogenic/adipogenic differentiations and osteoclastogenesis, respectively. Our data illustrated that ELP had a significant preventive effect against bone loss induced by tail-suspension (TS) at day 28 (p < 0.01) as indicated in the reduction in BMD loss and the preservation of bone micro-architecture. ELP could significantly promote the osteogenesis and suppress the adipogenesis (p < 0.05) in MSCs. Besides, significant inhibition of osteoclast formation (p < 0.01) was found in ELP-treated RAW264.7 cells upon receptor activator of nuclear factor kappa-B ligand induction. Our study presents the first scientific evidence that ELP had a significant preventive effect against bone loss induced by TS through the actions of enhancing osteogenesis, suppressing adipogenesis and osteoclastogenesis. Copyright © 2012 John Wiley & Sons, Ltd.
Phytotherapy Research 05/2012; · 2.09 Impact Factor
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Xiao-Qiang Han,
Ben Chung Lap Chan,
Cai-Xia Dong,
Yin-Hua Yang,
Chun-Hay Ko,
Grace Gar-Lee Yue,
Dan Chen,
Chun-Kwok Wong, Clara Bik-San Lau,
Peng-Fei Tu,
Pang-Chui Shaw,
Kwok-Pui Fung,
Ping-Chung Leung,
Wen-Luan Hsiao,
Quan-Bin Han
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ABSTRACT: A polysaccharide (GSP-6B) with a molecular mass of 1.86 × 10⁶ Da was isolated from the fruiting bodies of Ganoderma sinense . Chemical composition analysis, methylation analysis, infrared spectroscopy, and nuclear magnetic resonance spectroscopy were conducted to elucidate its structure. GSP-6B contains a backbone of (1→6)-linked-β-D-glucopyranosyl residues, bearing branches at the O-3 position of every two sugar residues along the backbone. The side chains contain (1→4)-linked-β-D-glucopyranosyl residues, (1→3)-linked-β-D-glucopyranosyl residues, and nonreducing end β-D-glucopyranosyl residues. An in vitro immunomodulating activity assay revealed that GSP-6B could significantly induce the release of IL-1β and TNF-α in human peripheral blood mononuclear cell (PBMC) and showed no toxicity to either PBMC or a human macrophage cell line THP-1. GSP-6B could also activate dendritic cells (DC) by stimulating the secretion of IL-12 and IL-10 from DC.
Journal of Agricultural and Food Chemistry 04/2012; 60(17):4276-81. · 2.82 Impact Factor
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ABSTRACT: Multidrug resistance is a major problem in hepatocellular carcinoma. Hedyotiscone A, a compound isolated from Chinese herbal medicine Hedyotis corymbosa (HC, family Rubiaceae), was used as the chemical marker to distinguish between HC and an anticancer herb Hedyotis diffusa (HD) in our previous study. The present study aimed to investigate whether HA exhibited antiproliferative activities in multidrug-resistant hepatocellular carcinoma cells R-HepG2 and the parental cells HepG2 using MTT assay and [(3)H]-thymidine incorporation assay. Our results showed that HA could significantly inhibit cell proliferation in R-HepG2 and HepG2 (IC(50) = 43.7 and 56.3 µg/mL, respectively), but not in normal human liver cells WRL-68 (IC(50) > 100 µg/mL) cells, suggesting its selective cytotoxic effects. Besides, HA induced apoptosis in R-HepG2 cells, as confirmed by annexin-V & propidium iodide staining, and DNA fragmentation assay. The caspase cascade was activated as shown by a significant increase of cleaved caspases-3, -7 and -9 in HA-treated R-HepG2 cells. The activities and protein expression of P-glycoprotein as well as mRNA expression of MDR1 were also decreased in HA-treated R-HepG2 cells. Our study demonstrated for the first time the antiproliferative activities of hedyotiscone A in multidrug-resistant R-HepG2 cells. The findings revealed the potential of this compound in treating multidrug-resistant tumor.
Xenobiotica 02/2012; 42(6):562-70. · 1.79 Impact Factor
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Kit-Man Lau,
Kwok-Kin Lai,
Cheuk-Lun Liu,
Jacqueline Chor-Wing Tam,
Ming-Ho To,
Hin-Fai Kwok,
Ching-Po Lau,
Chun-Hay Ko,
Ping-Chung Leung,
Kwok-Pui Fung,
Simon Kar-Sing Poon, Clara Bik-San Lau
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ABSTRACT: Astragali Radix (AR) and Rehmanniae Radix (RR) are two traditional Chinese medicines widely used in China for treating diabetes mellitus and its complications, such as diabetic foot ulcer.
In our previous study, a herbal formula NF3 comprising AR and RR in the ratio of 2:1 was found effective in enhancing diabetic wound healing in rats through the actions of tissue regeneration, angiogenesis promotion and inflammation inhibition. The aims of the present study were to investigate the herb-herb interaction (or the possible synergistic effect) between AR and RR in NF3 to promote diabetic wound healing and to identify the principal herb in the formula by evaluating the potencies of individual AR and RR in different mechanistic studies.
A chemically induced diabetic foot ulcer rat model was used to examine the wound healing effect of NF3 and its individual herbs AR and RR. For mechanistic studies, murine macrophage cell (RAW 264.7) inflammation, human fibroblast (Hs27) proliferation and human endothelial cell (HMEC-1) migration assays were adopted to investigate the anti-inflammatory, granulation formation and angiogenesis-promoting activities of the herbal extracts, respectively.
In the foot ulcer animal model, neither AR nor RR at clinical relevant dose (0.98g/kg) promoted diabetic wound healing. However, when they were used in combination as NF3, synergistic interaction was demonstrated, of which NF3 could significantly reduce the wound area of rats when compared to water group (p<0.01). For anti-inflammation and granulation formation, AR was more effective than RR in inhibiting lipopolysaccharide (LPS)-induced nitric oxide production from RAW 264.7 cells and promoting Hs27 fibroblast proliferation. In the aspect of angiogenesis promotion, only NF3 promoted cell migration of HMEC-1 cells.
AR plays a preeminent role in the anti-inflammatory and fibroblast-proliferating activities of NF3. The inclusion of RR, however, is crucial for NF3 to exert its overall wound-healing as well as the underlying angiogenesis-promoting effects. The results of present study justified the combined usage of AR and RR in the ratio of 2:1 as NF3 to treat diabetic foot ulcer and illustrated that AR is the principal herb in this herbal formula.
Journal of ethnopharmacology 02/2012; 141(1):250-6. · 2.32 Impact Factor
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Chun-Hay Ko,
Wing-Sum Siu,
Hing-Lok Wong,
Si Gao,
Wai-Ting Shum,
Ching-Po Lau,
Sau-Wan Cheng,
Jacqueline Chor-Wing Tam,
Leung-Kim Hung,
Kwok-Pui Fung, Clara Bik-San Lau,
Quan-Bin Han,
Ping-Chung Leung
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ABSTRACT: Antiresorptive drugs, alendronate and raloxifene, are effective in lowering bone mineral density (BMD) loss in postmenopausal women. However, long-term treatment may be associated with serious side effects. Our research group has recently discovered that a Chinese herbal formula, ELP, could significantly reduce BMD loss in animal and human studies. Therefore, the present study aimed to investigate the potential synergistic bone-protective effects of different herb-drug combinations using ovariectomized rats. To assess the efficacy of different combinations, the total BMD was monitored biweekly in the 8-week course of daily oral treatment. Bone microarchitecture, bone strength, and deoxypyridinoline level were also determined after 8 weeks. From our results, coadministration of ELP and raloxifene increased the total tibial BMD by 5.26% (2.5 mg/kg/day of raloxifene; P = 0.014) and 5.94% (0.25 mg/kg/day of raloxifene; P = 0.026) when compared with the respective dosage groups with raloxifene alone. Similar synergistic effects were also observed in BMD increase at distal femur (0.25 mg/kg/day; P = 0.001) and reduction in urinary deoxypyridinoline crosslink excretion (2.5 and 0.25 mg/kg/day; both P = 0.02). However, such interactions could not be observed in all alendronate-treated groups. Our data provide first evidence that ELP could synergistically enhance the therapeutic effects of raloxifene, so that the clinical dosage of raloxifene could be reduced.
Evidence-based Complementary and Alternative Medicine 01/2012; 2012:203732. · 4.77 Impact Factor
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ABSTRACT: Both danshen (D) and gegen (G) have proven relaxant effects on vascular smooth muscle, thus their potential bladder inhibitory effects have impending interests in urology. The aim of this study was to demonstrate the novel effects of D and G on detrusor smooth muscle contractility. Urothelium-intact (+UE) and urothelium-denuded (-UE) detrusor strips were isolated from the rat. Isometric tension was measured using a myograph system. Carbachol (CCh) was used to pre-contract the detrusor strips prior to stepwise relaxation by adding extracts of D, G, and a DG (7:3) formulation. Tonic relaxation level and phasic contractile activity under the herbal treatments were analyzed. There was no difference in the herbal effects between +UE and -UE strips. D alone induced a much smaller relaxation than G alone or DG. G alone also suppressed phasic amplitude but not phasic frequency while DG suppressed both parameters. D and G acted synergistically to yield the observed effects on detrusor smooth muscle. The findings showed that the DG formulation were able to relax the detrusor as well as suppress phasic contractions, both actions important in maintaining normal bladder filling and urine storage processes. Hence DG may have new application in the management of bladder disorders.
International Journal of Physiology, Pathophysiology and Pharmacology 01/2012; 4(1):36-44.
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Ben Chung-Lap Chan,
Hua Yu,
Chun-Wai Wong,
Sau-Lai Lui,
Claude Jolivalt,
Carine Ganem-Elbaz,
Jean-Marc Paris,
Barbara Morleo,
Marc Litaudon, Clara Bik-San Lau,
Margaret Ip,
Kwok-Pui Fung,
Ping-Chung Leung,
Quan-Bin Han
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ABSTRACT: Bacterial resistance to antibiotics has become a serious problem of public health that concerns almost all currently used antibacterial agents and that manifests in all fields of their application. To find more antibacterial agents from natural resources is all the time considered as an important strategy. Sophora flavescens is a popularly used antibacterial herb in Chinese Medicine, from which prenylated flavones were reported as the antibacterial ingredients but with a major concern of toxicity. In our screening on the antibacterial activities of various chemicals of this herb, 18 fractions were obtained from 8 g of 50% ethanol extract on a preparative high-speed counter-current chromatography (HSCCC, 1000 ml). The system of n-hexane/ethyl acetate/methanol/water (1:1:1:1) was used as the two-phase separation solvent. A chalcone named kuraridin was isolated from the best anti-MRSA fraction, together with sophoraflavanone G, a known active ingredient of S. flavescens. Their structures were elucidated by analysis of the NMR spectra. Both compounds exhibited significant anti-MRSA effects, compared to baicalein that is a well known anti-MRSA natural product. More important, kuraridin showed no toxicity on human peripheral blood mononuclear cells (PBMC) at the concentration up to 64 μg/ml while sophoraflavanone G inhibited over 50% of cellular activity at 4 μg/ml or higher concentration. These data suggested that opening of ring A of the prenylated flavones might decrease the toxicity and remain the anti-MRSA effect, from a viewpoint of structure-activity relationship.
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 12/2011; 880(1):157-62. · 2.78 Impact Factor
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Qi Zhang,
Fan Wei,
Chi Chun Fong,
Wai Kin Yu,
Yao Chen,
Chi Man Koon,
Kit Man Lau,
Ping Chung Leung, Clara Bik San Lau,
Kwok Pui Fung,
Mengsu Yang
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ABSTRACT: The herbs Astragali Radix (AR) and Rehmanniae Radix (RR) have long been used in traditional Chinese Medicine and serve as the principal herbs in treating diabetic foot ulcer.
Chinese herbal formulus comprising Astragali Radix (AR) and Rehmanniae Radix (RR) have been shown to improve the healing of diabetic foot ulcer through enhancing the viability of primary fibroblasts in diabetic patients suffering insulin resistance. Our previous study demonstrated that the herbal formula NF3 comprising of AR and RR in the ratio of 2:1 was effective in promoting wound healing in diabetic rats, and in vitro data indicated that the wound healing effects of NF3 might be due to the regulation and coordination of inflammation, angiogenesis and tissue regeneration. However, the underlying molecular mechanism has not been well investigated. In this study, we investigated the cellular and molecular effects of the herbal formula NF3 on human skin fibroblast cells.
Human skin fibroblast cells Hs27 were treated with NF3 ranging from 0 to 8 mg/ml for 24h, and the cells without NF3 treatment were used as control. Cell proliferation assay and cell cycle analysis were performed. Transcriptional profiles of Hs27 cells upon NF3 treatment were acquired by using a human cDNA microarray containing 10,000 genes, and the signaling pathways differentially regulated by NF3 were identified and analyzed.
NF3 promoted Hs27 cell proliferation and cell cycle progression. Microarray analysis revealed that 116 genes were differentially expressed upon NF3 treatment. Functional analysis of the genes indicated that NF3 mainly activated Wnt and angiogenesis related pathways, which are directly related to cell proliferation, angiogenesis, extracellular matrix (ECM) formation and inflammation during the process of wound healing.
This study provides insight into the molecular mechanism of how the herbal formula Astragali Radix and Rehmanniae Radix may serve as potential therapeutics for wound healing.
Journal of ethnopharmacology 10/2011; 138(3):668-75. · 2.32 Impact Factor
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ABSTRACT: Green tea has been demonstrated recently as a potent bone supportive agent. Our previous studies showed that green tea and its polyphenolic constituents can promote bone-forming osteoblast activities and inhibit the bone-resorpting osteoclast formation. The objective of the present study was to investigate whether green tea and its components can regulate the osteogenic and adipogenic differentiation in pluripotent rat mesenchymal stem cells (MSCs). The rat MSCs were isolated from the bone marrow of tibiae and femora. The cells were treated with decaffeinated green tea extract (GTE) and six tea polyphenols under osteogenic induction. The alkaline phosphatase (ALP) activities and matrix calcium (Ca) deposition were assessed after 7 and 14 days of treatment. Our results demonstrated that GTE could significantly increase ALP dose dependently in the concentrations without cytotoxicity (0-100 μg/mL). Among six tested tea polyphenols, epigallocatechin (EGC) was shown to be the most effective in promoting osteogenic differentiation. At 20 μM, EGC increased ALP levels and Ca deposition significantly by 2.3- and 1.7-fold, respectively, when compared with the control group. EGC also increased the mRNA expression of bone formation markers runt-related transcription factor 2, ALP, osteonectin, and osteopontin. Furthermore, EGC demonstrated its antiadipogenicity by decreasing the adipocyte formation and inhibiting the mRNA expression levels of the adipogenic markers peroxisome proliferator-activated receptor γ, ccaat/enhancer-binding protein β, and fatty acid binding protein 4. In conclusion, this is the first report of the dual action of green tea polyphenol EGC in promoting osteogenesis and inhibiting adipocyte formation in MSCs. Our results provide scientific evidence to support the potential use of green tea in supporting the bone against degenerative diseases such as osteoporosis.
Journal of Agricultural and Food Chemistry 09/2011; 59(18):9870-6. · 2.82 Impact Factor
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ABSTRACT: Danshen (root of Salvia miltiorrhiza) and Gegen (roots of Pueraria lobata) are traditional Chinese medicines that have been used in combination for cardiovascular disease treatment.
The present study was performed to investigate the effect of Danshen-Gegen decoction on rat myocardium cell line H9c2 and the possible molecular mechanisms.
Rat heart myocardium H9c2 cells were treated with or without Danshen-Gegen decoction (DG) ranging from 10 to 1000μg/ml for 24h. Cell viability was measured by Alarma blue assay and cell proliferation assay was performed by BrdU Cell Proliferation ELISA kit. The activation of mitogen-activated protein kinase and insulin pathways was analyzed by Luminex technology and the growth factors and cytokine expression of H9c2 cells induced by DG was evaluated by protein array. Moreover, a rat functional specific cDNA microarray was constructed to study the gene expression profiles of H9c2 cells upon the DG treatment at 50μg/ml for 24h.
DG promoted H9c2 cell viability and cell proliferation at dose-dependent manner within the range between 0 and 250μg/ml. A Bio-Plex assay kit (Bio-Rad Bioscience) was used to detect the expression level of phosphoprotein as well as total proteins involved in the MAPK and insulin pathways. Significant phosphorylation of ERK, c-Jun, JNK, p38, AKT, IGF-IR, IRS-1and I kappa B were observed after DG treatment at 2h or 4h. A rat cytokine antibody array was used to detect and quantify 22 growth factors and cytokines in samples collected from the control and DG treated H9c2 cells. In the category of growth factors, GM-CSF, CNIF and b-NGF were stimulated by DG, while the expression of TIMP-1 was suppressed. For cytokine expression, it was found that DG stimulated three interleukin subclasses, IL-1α, 1X and 6, respectively. However, the expression of pro-inflammatory factors such as TNF-α and IFN-γ were down-regulated significantly. Moreover, the microarray analysis revealed that DG significantly up-regulated anti-apoptosis related genes such as Cdkn2c and Ppp3ca, and several cardiovascular disease suppressers and anti-inflammatory mediators; on the other hand, pro-apoptotic related genes including Caspase and Tnf-α were down-regulated by DG. Based on the results, a tentative scheme was proposed to show that the activation of the MAPK and insulin pathways are involved in the bioactive effect of Danshen-Gegen decoction on cardiomyocytes.
Our study suggested that Danshen-Gegen decoction has proliferative effect on myocardium cells via MAPK and insulin signaling pathways. The molecular mechanism of the action may include the up-regulation of IRS/AKT and JNK pathways as well as the inhibition of TNF and p38 pathways.
Journal of ethnopharmacology 09/2011; 138(1):60-6. · 2.32 Impact Factor
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Cheuk-Lun Liu,
Ling Cheng,
Hin-Fai Kwok,
Chun-Hay Ko,
Tai-Wai Lau,
Chi-Man Koon,
Ming Zhao,
Ching-Po Lau,
Kit-Man Lau,
Chun-Wai Wong,
Ping-Chung Leung,
Kwok-Pui Fung, Clara Bik-San Lau
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ABSTRACT: The root of Rehmannia glutinosa (RR) is commonly used as a wound-healing agent in various traditional Chinese herbal formulae; while angiogenesis is one of the crucial aspects in wound-healing.
The objective of the present study was to investigate the angiogenesis effects of RR aqueous crude extract and its active component(s) using zebrafish model.
The in vivo angiogenesis effect was studied using transgenic TG(fli1:EGFP)(y1)/+(AB) zebrafish embryos by observing the capillary sprouts formation in sub-intestinal vessel (SIV) of zebrafish embryos after 72 h post-fertilization under fluorescence microscopy.
Our results indicated that RR aqueous crude extract (250 μg/ml) exhibited significant angiogenesis effect, with an increase in capillary sprouts formation in SIV. Following sequential solvent partition of the RR aqueous crude extract with dichloromethane, ethyl acetate and n-butanol successively, the dichloromethane fraction (DCM) was found to have the most sprouts formation in the SIV region. Subjected to column chromatography, DCM fraction was further fractionated into six sub-fractions and among these tested, the sub-fraction C2 exhibited the most potent angiogenesis effect. The major component, C2A, was isolated and identified as norviburtinal using nuclear magnetic resonance (NMR) and mass spectrometry (MS). The compound norviburtinal (at 50 μg/ml) was shown to possess significant angiogenesis effect in zebrafish model (p < 0.001).
Norviburtinal was, for the first time, found in the extract of RR and possessed novel angiogenesis effect. Bioassay-guided fractionation suggested that norviburtinal was not the only active component responsible for the angiogenesis effect of RR.
Journal of ethnopharmacology 08/2011; 137(3):1323-7. · 2.32 Impact Factor
