Journal of Medicinal Plant Research

Online ISSN: 1996-0875
Publications
Article
Cimicifugeae is one of the rich sources for various active components and the health promoting and therapeutic values of the components have been corroborated by long-term use in folk medicine and traditional Chinese medicine. Increasing interest in Cimicifugeae pharmaceutical resources has led to the further discoveries of triterpenoid saponins, phenolic compounds, chromones, and many other compounds in various species of Cimicifugeae, and to the investigations on their chemotaxonomy, molecular phylogeny, and bioactivities. Based on our pharmacophylogenetic studies, the progress in phytochemistry, chemotaxonomy, molecular biology, and phylogeny of Cimicifugeae had been summarized since 2007, especially Cimicifuga L. ex Wernisch. and Actaea L., and their relevance to therapeutic efficacy. An exhaustive literature survey is used to characterize the global scientific effort in the phytochemical and biological studies of Cimicifugeae. More triterpenoid saponins have been found in various species, among which the cimigenol type (type A) is predominant. The versatile bioactivities of saponins and extracts, as well as those of phenolics and other ingredients, were summarized and discussed. The morphology-based five-genus classification of Cimicifugeae is not supported by molecular phylogeny. Molecular phylogeny based on nuclear and chloroplast DNA sequences tends to merge Cimicifuga Wernisch., Souliea Franch., and Actaea L. into a single genus. It is indispensable to integrate the emerging technologies into Cimicifugeae research for both the sustainable utilization of Cimicifugeae pharmaceutical resources and finding novel compounds with potential clinical utility and less adverse effects. Systems biology and omics technologies would play an increasingly important role in booming pharmaceutical research involving bioactive compounds of Cimicifugeae.
 
Variance analysis of the effect of 1000 grain weight on sprouting of Salvia L.
Comparison of studied traits.
Article
The harmful effects of chemicals and the side effects of chemical drugs on human health have a widely focused global attention on herbal drugs and medicinal plants. The increasing use of medicinal plants in the world is more than enough to show the significance of cultivating and producing such plants. Grains of most medicinal plants have very long dormancy periods, so shortening the dormancy period and increasing sprouting rates through laboratory methods may be effective in the proliferation of medicinal plants. Salvia L. is a member of the family Lamiaceae with more than 900 species all over the world. One of the major standards of seed quality is the 1000 grain weight, which is effective on sprouting, seed potential, seedling growth, and plant performance. Therefore, an experiment was run in the form of completely random blocks in 3 iterations to study the effects of the 1000 grain weight on the sprouting rate and seed potential of Salvia L. Ten different 1000 seed weight treatments were used in a standard sprouting test and accelerated aging test inside a germinating machine. The experiments were run in three iterations, each including 50 seeds based on rules set forth by the International Seed Testing Association. The experiments tended to analyze seed properties such as sprouting percentage, seedling length, and weight of dried seedling. Obtained data was analyzed by SAS software. Results indicated that in the standard sprouting test, the 1000 grain weight had significant effects on growth traits such as length and weight of dry seedling, yet it had no significant effect on sprouting percentage. In addition, in the accelerated aging test the 1000 seed weight had a significant effect on sprouting percentage and dry seedling weight, while there was no significant effect on seedling length.
 
The effect of peroxyl radical scavenging activities (Trolox equivalent, M) of PSE-1 different concentrations in vitro system. The oxygen radical absorbance capacity (ORAC) value is calculated by dividing the area under the sample curve by the area under the Trolox curve, with both areas being corrected by subtracting the area under the blank curve. One ORAC unit is assigned as the net area of protection provided by Trolox at a final concentration of 1 M. The area under the curve for the sample is compared to the area under the curve for Trolox, and the anti-oxidative value is expressed in micromoles of Trolox equivalent per liter. The results represent the mean ± SE. of values obtained from three measurements. Different corresponding letters indicate significant differences at p < 0.05 by Duncan's multiple range test. Kim et al. (2007). 
The effect of supplementation in vitro with different concentrations of PSE-1 on 200 μM H2O2 induced DNA damage in human leukocytes. NC, 1% DMSO (without oxidative stimulus) treated negative control; PC, 200 μM H2O2 treated positive control. The results represent the mean ± SE. of values obtained from three measurements. Different corresponding letters indicate significant differences at p < 0.05 by Duncan's multiple range test. Kim et al. (2007). 
Effect of PSE-1 on glutamate-induced cytotoxicity in N18-RE-105 cells. (A) MTT reduction assay. (B) LDH release assay. Values are mean with standard error of triplicate experiments. Cells were pretreated with the indicated concentration of PSE-1 for 30 min, and then further treated with 20 mM glutamate for 24 h. The MTT reduction rate was the difference in absorbance values between the treated and control wells divided by the control absorbance. The LDH release rate was presented as percentage of control values. As the results of LDH release assay, data were normalized to the activity of LDH released from vehicle-treated cells (100%) and expressed as percentage of the control (obtained from separate plating). Kim et al. (2007). 
Effects of PSE-1 on glutamate-induced morphologic changes in N18-RE-105. (a) Control; (b) N18RE-105 cells exposed to 20 mM of glutamate for 24 h; (c) N18-RE-105 cells treated with 100 µM PSE-1 for 30 min before exposure to 20 mM of glutamate for 24 h; (d) N18RE-105 cells treated with 250 μM PSE-1 for 30 min before exposure to 20 mM of glutamate for 24 h. Photographs were taken with a phase-contrast microscope at magnification × 100 and show that glutamate treatment decreased the number of viable cells and induced shrinkage and aggregation of cell bodies, whereas PSE-1 pretreatment attenuated the effects of glutamate treatment. Kim et al. (2007). 
Inhibition of glutamate-induced apoptosis in N18-RE-105 cells by PSE-1. Cells were preincubated with PSE-1 at the indicated concentrations for 30 min. The cells were then stimulated with 20 mM of glutamate for 24 h and then used for cell staining with Hoechst 33342. (a) Control; (b) 20 mM of glutamate; (c) 20 mM glutamate + PSE-1 100 μM; (d) 20 mM glutamate + PSE-1 250 μM. The nuclear morphology of cells was examined by flourescence microscopy (magnification × 400). Kim et al. (2007). 
Article
Neurodegenerative conditions, such as the Alzheimer and Parkinson diseases, are associated with the production of reactive oxygen species and resultant oxidative stress. Glutamate is the major excitatory neurotransmitter of the central nervous system and may induce cytotoxicity through persistent activation of glutamate receptors and through oxidative stress mechanisms. On the basis of this information, we established a screening system using N18-RE-105 cells to identify therapeutic agents that can protect cells from glutamate toxicity. During the course of our screening program, we recently isolated an active compound, 8,13-dihydroxy-9,11-octadecadienoic acid (PSE-1), from peanut sprouts, which prevents glutamate-induced cell death. The chemical structure of PSE-1 was identified using spectroscopic methods and by comparison with the value in the literature. The antioxidant and neuroprotective effects of PSE-1 were evaluated using the oxygen radical absorbance capacity assay, Comet assay, the 3-(4,5-dimethylthiazol-2-yl)-2,5,-diphenyltetrazolium bromide reduction assay, the lactate dehydrogenase release assay, a morphological assay and Hoechst 33342 staining. The results of the assays demonstrate that PSE-1 has neuroprotective effects and that PSE-1 could be a new potential chemotherapeutic agent against neuronal diseases.
 
calculated dipole moment μ (Debye) versus dielectric constant ( ɛ ) at bulk and five different solvents. 
Article
Quantum Monte Carlo (QMC), Molecular Dynamics (MD) simulations and Density Functional Theory (DFT) calculations at the level of B3LYP/3-21G carried out on the structure and stability of B 10 N 11 H 7 (Thr) 2 in bulk and different solvent medium. NMR parameters and thermodynamic properties were calculated to obtain chemical shift, stability and solvent effect. It was found that computationally efficient solvent modeling is possible and can reveal fine details of molecular structure, stability and dynamics. The results showed high stability of B 10 N 11 nanocone, it can be the best candidate and much favorable in biological systems and drug delivery. Key words: Molecular dynamics simulations, solvent effect, DFT, B 10 N 11 H 7 (Thr) 2 , nanocone, Monte Carlo.
 
The main HMBC correlations of Euphorbia factor L1 (HC).
Article
In this article, we reported the isolation and characterization of Euphorbia factor L1 from Caper Euphorbia seed. The complete 1 H and 13 C NMR signals were assigned with the help of 1D, 2D NMR techniques. In addition, the anticancer activities of Euphorbia factor L1 were investigated. It exhibited potent cytotoxicity to KB, KBv200, MCF-7 and MCF-7/ADR cells with the IC 50 values of 30.83 ± 2.93, 28.11 ± 3.08, 39.47 ± 4.03 and 42.69 ± 4.27 g/ml, respectively.
 
Metabolic profile of different 14 C substrates and 14 C content into primary metabolic fractions into root of catharanthus. All values in (Bq/g. F. mass root).
Article
Studies were performed using 14 C radiolabel to elucidate the extent to which carbon from four sources (CO 2 , sucrose, glucose and acetate) contribute to the primary metabolites and alkaloids produced by the roots, stems and leaves of Catharanthus roseus plant. 14 CO 2 derived metabolites were most preferred for biosynthesis of leaf and root alkaloids. Roots were the major accumulators of metabolites accompanied by higher 14 C content into total alkaloids. 14 C-glucose derived metabolites were the second most preferred substrate for leaf and stem alkaloid accumulation and the stem showed highest incorporation into alkaloids. 14 C-sucrose was the third most preferred carbon source for biosynthesis of leaf, stem and root alkaloids. Among these substrates, 14 C-acetate was the least preferred by the plant for biosynthesis of alkaloids of leaf, stem and root. There were variations in the metabolite mobilization of sugars, amino acids and organic acids between leaf, stem and root when different C sources were supplied. A floating metabolic pool seems to influences the alkaloid biosynthesis occurring in leaf, stem and root. The relative higher incorporation of 14 CO 2, 14 C-sucrose and 14 C-glucose suggests the preferential utilization of metabolites from MEP/terpenoid pathway that contribute to alkaloid accumulation as compared to 14 C acetate suggesting lower contribution by mevalonate/terpenoid pathway derived metabolites.
 
Article
IC 50 for a novel antibiotic against mouse hepatoma H 22 was determined to be 0.019 μg/mL by MTT assays. In KM mice bearing this kind of tumor that was treated by s.c. injection of antibiotic BS, marked cures were obtained. In terms of tumor weight, the treated/control=0.35. The conclusion is that BS merit further investigation as a potential anti-cancer candidate drug.
 
Article
Zingiberaceae plants exhibit various biological activities, but scientific knowledge of the plants in antimetastatic aspects is still very limited. The objective of this study is to examine the anti-metastatic potentials of 30 crude extracts from ten selected local Zingiberaceae species on hormone-independent, highly metastatic human breast cancer cells, MDA-MB-231. The Zingiberaceae rhizomes were extracted with petroleum ether, chloroform, and methanol using Soxhlet extractor system. Effects of the 30 extracts on proliferation and migration of MDA-MB-231 cells were evaluated using 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and scratch wound assay, respectively. Besides cytotoxicity, it is also vital to determine the potential toxicity of the extract in order to select the most promising extracts for anti-cancer drugs development. Thus, a special parameter–Selectivity Index (SX) was utilized as the selectivity indicator of tested extracts towards tumour cells. The results revealed that petroleum ether extracts of Alpinia galanga, Boesenbergia rotunda, and Zingiber zerumbet; as well as chloroform extracts of Alpinia galanga, Boesenbergia rotunda, and Curcuma domestica were screened to possess the most effective anti-proliferation and anti-migration activities against MDA-MB-231 cells. Three most desirable extracts were selected based on both their anti-metastatic potentials and SX, and were subjected for qualitative analysis using thin-layer chromatography (TLC).
 
Article
At least 15 elements can be considered as an essential nutrients for human health but their toxic concentration should also be considered for preparation of herbal medicines. 24 medicinal plants were selected for metals analysis renowned for their therapeutic medicinal use. The metals analyzed include sodium (Na), potassium (K), lithium (Li), calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), lead (Pb), manganese (Mn), cupper (Cu), nickel (Ni), chromium (Cr) and cadmium (Cd) by flame emission spectroscopy and atomic absorption spectroscopy. It was investigated that the level of essential elements was higher as compared to trace elements as Zn, Pb, Mn, Cu, Ni, Cr and Cd were not detected, depicting their lesser concentration in selected plants. The highest concentration (ppm) of macro nutrients Na (792), Li (78) were found in Gauzuban (Onosma bracteatum ); and Ca (164), K (180 found in Dandelion (Taraxacum officinale ); and that of Mg (11.5) both in Neem ( Azardirachta indica ) and Gainda (Tagetes minuta ). The results also showed that Methi ( Trigonella foenum-graceum ) has highest concentration of Fe (29.8); Gainda ( T. minuta ) has highest concentration of Zn (29.8). Presence of different metals in these medicinal plants was correlated to their medicinal use.
 
Article
At least 15 elements can be considered as an essential nutrients for human health but their toxic concentration should also be considered for preparation of herbal medicines. 24 medicinal plants were selected for metals analysis renowned for their therapeutic medicinal use. The metals analyzed include sodium (Na), potassium (K), lithium (Li), calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), lead (Pb), manganese (Mn), cupper (Cu), nickel (Ni), chromium (Cr) and cadmium (Cd) by flame emission spectroscopy and atomic absorption spectroscopy. It was investigated that the level of essential elements was higher as compared to trace elements as Zn, Pb, Mn, Cu, Ni, Cr and Cd were not detected, depicting their lesser concentration in selected plants. The highest concentration (ppm) of macro nutrients Na (792), Li (78) were found in Gauzuban (Onosma bracteatum); and Ca (164), K (180 found in Dandelion (Taraxacum officinale); and that of Mg (11.5) both in Neem (Azardirachta indica) and Gainda (Tagetes minuta). The results also showed that Methi (Trigonella foenum-graceum) has highest concentration of Fe (29.8); Gainda (T. minuta) has highest concentration of Zn (29.8). Presence of different metals in these medicinal plants was correlated to their medicinal use.
 
Minimum inhibition concentration. 
Minimum fungicidal concentration. 
MIC and MFC results of fluoconazole. 
Article
3β-hydroxyllup-20(29)-en-28-oic acid (Betulinic acid) a Pentacyclic lupane triterpene known for anti-HIV activity and cytotoxic activity against various malignant versus nonmalignant cancer cell lines, was isolated from malaleuca bracteata. The structure was elucidated on the basis of spectroscopy analysis, including 2D-NMR correlation spectroscopy (COSY), nuclear overhauser enhancement spectroscopy (NOESY), heteronuclear multiple bond correlation (HMBC) and heteronuclear single quantum correlation (HSQC) experiments. The inhibitory zone (mm) ranges from 20 ± 0.02 to 30 ± 0.01 against the test organisms. Trichophyton tonsurans was the most sensitive organism (30 ± 0.01) which was observed to be greater than the standard drugs Fluoconazole (14 ± 0.90), Nystatin (17 ± 0.03) and Fulcin (23 ± 0.50) used as positive control. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) results indicated that a concentration of 0.625 and 2.5 mg/ml inhibited and completely kill Candida guielemondi, Candida stelletoidea, Candida parapsilosis, Candida pseudotropicalis, T. tonsurans and Microsporum canis except T. tonsurans which was kill at 1.25 mg/ml. While the rest of the organisms were inhibited and kill by a concentration of 1.25 and 2.5 mg/ml respectively, the results shows that the compound have great potential as antifungal drug.
 
Article
The cultured mycelium of strain Paecilomyces militaris CMG 01 (Pm) was extracted sequentially by hexane, ethyl acetate (EtOAc) and butanol (BuOH). Among them, the BuOH extract exhibited potent anti-proliferative activity on B16/F10 mouse melanoma and HT-29 human colon cancer cells. Active chemical fractions of the Pm BuOH extract led to the isolation of a nucleoside compound, which structure was identified as cordycepin on the basis of 1 H, 13 C NMR and MS data. The further evaluated on cordycepin showed that cordycepin significantly inhibited B16/F10 and HT-29 cells proliferation and its content in BuOH extract was 68.89 mg/g. our results suggest that the BuOH extract of Pm is a potential source of natural anticancer agents.
 
Article
Yacon (Smallanthus sonchifolius) tubers and leaves have been used widely as foodstuff and as remedy for urinary ailments, muscle pain, hyperlipidemia and diabetes mellitus. Recent studies have investigated on isolating active components for their anti-cancer potential against melanoma, cervical cancer and colon cancer. In this study, the cytotoxicity potential of hexane, methanol and DCM extracts of yacon leaves was assessed against MCF-7 (breast cancer), HT-29 (colon cancer) and HDFn (normal human dermal fibroblast) cell lines by using AlamarBlue® assay. Results showed significant reduction in cellular viability of MCF-7 cell lines caused by hexane, methanol and DCM extracts in a dose dependent manner, with DCM being the most potent. The DCM extract also produced significant cytotoxic activity against HT-29 cells, with IC50 lower than 5-fluorouracil. Effect on HDFn showed that three yacon extracts produced significantly lower cytotoxicity compared to drug controls with the DCM extract showing the least toxicity. Key words: Yacon, alamar, breast, colon, cancer, MCF-7, HT-29..
 
Article
Glucose deprivation, a feature of poorly vascularized solid tumors, activates the unfolded protein response (UPR) which is a stress-signaling pathway in tumor cells that is associated with the molecular chaperone, glucose-regulated protein 78 (GRP78). Induction of GRP78 protects cells against programmed cell death. Methanolic extract of Carthamus tinctorius (CTE) induced selective cytotoxicity in glucose-deprived HT-29 human colon carcinoma cells; this effect was not seen under normal growth conditions. CTE also suppressed the accumulation of the GRP78 protein and was highly toxic in HT-29 cells, with IC50 values for colony formation of < 50 μg/ml under 2-deoxyglucose (2DG) supplemented and glucose-deprived conditions. Interestingly, apoptotic activity of CTE was also detected by Hoechst staining and flow cytometric analysis. Therefore, these results suggest that CTE prevent the upregulation of GRP78 and exhibit selective cytotoxicity in glucose-deprived HT-29 cells.
 
Article
To observe the effects of electroacupuncture at Quchi point on carotid artery vascular smooth muscle cells (VSMCs) apoptosis of 2K1C-RHR and expression of Bcl-2 and Bax. The therapy methods in electroacupuncture group and electroacupuncture control group were carried out through the Quchi point and the WaiGuan point, respectively, on base of 2K1C-RHR model. The expression of Bcl-2 and Bax protein in carotid artery was detected by immunohistochemical method; Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect the apoptosis of carotid artery VSMCs. Compared with the normal control group, the apoptosis of carotid artery VSMCs of model control group increased, the expression of Bcl-2 increased and the expression of Bax decreased. The apoptosis rate of carotid artery VSMCs increased in the electroacupuncture group contrary to model control group (P<0.05), the expression of Bcl-2 decreased and the expression of Bax increased. There was not a significant difference in the apoptosis rate of carotid artery VSMCs between electroacupuncture control group and model control group (P>0.05). The vascular remodeling of hypertension rats in carotid artery can be influenced through the adjustment of VSMCs apoptosis by electroacupuncture.
 
Article
Phytochemical research on the aerial parts of Scutellaria barbata used variously in ethno medicinal treatments in southern China, resulted in the isolation of ethyl 4-hydroxy-3,5-dimethoxy-benzoate. Its structure was identified purely on the basis of spectroscopic analyses including EI-MS, IR and X-ray diffraction analysis. The crystal belonged to the monoclinic space group P2 (1)/c with a = 11.5521(6) Å, b = 13.5055(7) Å, c = 16.4171(7) Å, β = 117.240(3)°and Z = 8.This compound was firstly isolated from the genus Scutellaria.
 
Article
Effects of caffeic acid (CAF) and its derivatives on human liver CYP3A4 activity were evaluated by using diazepam as a substrate. It was found that CAF inhibited CYP3A4 activity by uncompetitive inhibition with IC50 0.72 µM, whereas ester and amide analogues inhibited CYP3A4 by competitive inhibition with IC50 value of 0.31, 0.37, 0.46, 0.49, 0.53, 0.58, 0.75 and 0.82 µM for ethyl 1-(3',4'-dihydroxyphenyl) propen amide (EDPA), phenmethyl 1-(3',4'-dihydroxyphenyl) propen amide (PMDPA), phenethyl 1-(3',4'-dihydroxyphenyl) propen amide (PEDPA), phenylethyl 1-(3',4'-dihydroxyphenyl) propenate (PC), ethyl 1-(3',4'-dihydroxyphenyl) propenate (EC), octyl 1-(3',4'-dihydroxyphenyl) propen amide (ODPA), octyl 1-(3',4'-dihydroxyphenyl) propenate (OC) and phenylmethyl 1-(3',4'-dihydroxyphenyl) propenate (BC), respectively. The K i values of CAF, PMDPA, EC, PC, ODPA, PEDPA, OC, EDPA and BC were 0.24, 0.29, 0.49, 0.56, 0.57, 0.59, 0.62, 0.62 and 1.03 µM, respectively. However, CAF and its derivatives had high potential to inhibit CYP3A4. Therefore, consumption of herbal medicine containing CAF and its derivatives that are concomitant with other medications should be cautiously monitored.
 
Article
The leaf of Nelumbo nucifera Gaertn has been used for summer heat syndrome as home remedy in China, and it has been used for the treatment of obesity in traditional medical clinics in China, so we investigated the pharmacological activity of the antiobesity effect of N. nucifera leaves extract. N. nucifera alkaloid (NNA), which is the main component in the N. nucifera leaves. 3T3-L1 preadipocytes and high fat diet (HFD)-induced obese SD rats were treated with NNA, and the effects of NNA on the apoptosis in 3T3-L1 preadipocytes and body weight, as well as plasma TC, TG and LDL-C levels in HFD-induced rats were investigated. The results demonstrated that NNA decreased cell population growth of 3T3-L1 preadipocytes and increased the apoptotic cells in a time-and dose-dependent manner. Furthermore, NNA reduced the body weight, the lee's index, adipose tissue weight, and plasma lipid levels in HFD-induced obese rats. Take together, we demonstrated that NNA can efficiently induces apoptosis in 3T3-L1 preadipocytes, and have further implication in in vivo antiobesity effect.
 
Article
The ethanol extract of the aerial parts of Scutellaria barbata used variously in ethno medicinal treatments in Korea and southern China was studied. A silica gel column chromatography of the ethanol extract using petroleum ether-acetone eluted a white slice crystal which was identified purely by spectral analyses as (6S, 9R)6-hydroxy-4,4,7a-trimethyl-5,6,7,7a-tetrahydro-1-benzofuran-2(4H)-one. The compound was firstly isolated from the genus S. barbata.
 
Article
This study was initiated to evaluate the lipid-lowering effect of Coriolus versicolor (CV) extracts in poloxamer 407 (P-407)-induced hypercholesterolaemic and high cholesterol-fed rats. In the chemically induced hypercholesterolaemic rat model, hypercholesterolaemia was induced by intraperitoneal administration of P-407 (500mg/kg). Serum lipid levels, including total cholesterol (TC), triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C) levels, increased significantly (p<0.001) in the P-407-induced hypercholesterolaemic rats as compared to the normal control rats. Interestingly, CV extract administration caused a significant reduction in serum TC (p<0.01) and TG (p<0.05) levels in P-407-treated rats, as well as a reduction in the coronary risk index (CRI) in these rats. In the diet-induced hypercholesterolaemic rat model, the rats were fed a high cholesterol diet containing (by weight) 2% cholesterol, 0.5% cholic acid and 10% butter. The serum TC and LDL-C levels of rats that were fed a high cholesterol diet increased significantly (p<0.001) after 14 days as compared to the rats fed a normal diet. Serum TC and LDL-C levels were significantly reduced in a dose-dependent manner in rats given CV extract orally and fed a high cholesterol diet for 14 days. The administration of CV extract also significantly increased high-density lipoprotein cholesterol (HDL-C) levels in rats fed a high cholesterol diet; in addition, CV extract reduced the CRI ratio. Hence, the 14 days administration of CV extract resulted in significant reductions in the lipid profiles of these rats compared to the untreated hypercholesterolemic rats. Our results suggest that the consumption of CV extract has the potential to reduce or prevent hypercholesterolaemia.
 
Article
The effects of two flavones (baicalein, chrysin) and two flavonols (galangin, kaempferol) on induction of differentiation in a human esophageal squamous cell carcinoma (KYSE-510) cell line were investigated in this study. Four compounds were found to be able to inhibit proliferation of KYSE-510 cells in a dose-and time-dependent manner. The inhibitory potency of them on the growth of KYSE-510 cells was in the order of galangin > > > > chrysin > > > > baicalein > > > > kaempferol. The KYSE-510 cells treated with these compounds for 24 h showed differentiation characteristics including the differentiation-specific morphological changes, the augmentation of differentiation markers and the inhibition of human telomerase reverse transcriptase. The potency of these compounds on induction of differentiation was similar to that of them on growth inhibition. To elucidate the possible mechanisms by which these compounds modulate proliferation and differentiation in KYSE-510 cells, the alterations of three genes (p21 waf1 , cyclin B1 and D1) related to proliferation and differentiation were investigated by real-time RT-PCR and Western blot. The results showed that the up-regulation of p21 waf1 and down-regulation of cyclin B1 and D1 at the mRNA and protein levels in KYSE-510 cells treated with four compounds were observed, which implied that p21 waf1 , cyclin B1 and D1 might be target genes of these compounds in inducing differentiation.
 
Article
Inducing leukemia cell apoptosis is a major therapeutic strategy. Herein we investigate the enhancing effect of the herbal constituent parthenolide on aclarubicin-induced apoptosis of human HL-60 leukemia cells. HL-60 cells were incubated with aclarubicin in the absence or presence of different doses of parthenolide. Apoptosis was assessed by flow cytometry using annexin V-propidium iodide double staining. To investigate the molecular mechanism by which parthenolide enhances aclarubicin-induced HL-60 apoptosis, caspase 3 and caspase 9 expression, as well as Cox-2 and NF-κB activity, were assessed by western blot. Following exposure to aclarubicin for 20 h, the percentage of cells undergoing apoptosis was highly correlated with dose. However, there was no significant apoptosis at a low aclarubicin concentration (0.1 µg/ml). Combined treatment at low aclarubicin and parthenolide concentration had a dose-dependent effect on apoptosis. In addition, combined parthenolide and aclarubicin treatment had a significant synergistic inhibitory effect on caspase 3, caspase 9 and Cox-2, NF-κB activity. In conclusion, parthenolide sensitizes leukemia cells to aclarubicin-induced apoptosis through increasing expression of cleaved caspase 3 and caspase 9, maybe by suppress Cox-2 and NF-κB activation, suggesting a new avenue of treatment of leukemia.
 
Article
Uyghur medicinal tea (UMT) is preferably used as a common nutritional additive by Uyghur population, especially by the aged people including centenarians from southern part of Xinjiang, along the Silk Road of China. The study showed an evidence for the antioxidant properties of UMT extract by using two different oxidative systems in vitro: (1) the myeloperoxidase/H 2 O 2 /HOCl enzymatic system in human HL-60 cell line and (2) the cytochrome P450/NADPH reductase system in rat liver microsomes. The EGB 761 Ginkgo biloba extract was used as the reference. Reactive oxygen species (ROS) production was measured by fluorescence of 2',7'-dichlorofluorescein (DCF) originated from 2',7'-dichlorofluorescein diacetate (DCFHDA) probe oxidation. UMT extract inhibited ROS formation by 81, 86 and 73% in the HL 60 cell line system (at the concentrations of 0.25, 0.50 and 0.75 mg/ml) and at similar rates in the microsomal system. In the same conditions, G. biloba extract inhibited ROS activity by 39, 78 and 81% in HL 60 cells, and by 78, 87 and 89% in the microsomal system. The present study shows that UMT possesses relatively high antioxidant activity, comparable to the free radical scavenger properties of EGB 761 extract. This effect could explain its benefic preventive action against age-related pathologies. Habitual consumption of UMT may provide beneficial effects in prevention of atherosclerosis and oxidative stress in aging process.
 
Article
Sinomenine (SN) a component from Sinomenium acutum, widely used as an anti-inflammatory agent in Chinese traditional medicine, was reported to have anti-tumor activities. This study aims to examine the anti-leukemic activity of SN with or without aclarubicin (Acla) on leukemia cells and its molecular mechanism. HL-60 cells were treated with SN (5 to 20 ng/ml) with or without Acla (0.05 to 0.25 µg/ml). To study apoptosis, the cells were stained with Annexin V -propidium iodide (PI) and assessed by flow cytometry. To investigate the molecular mechanism, Caspase 3, Caspase 9 and cyclooxygenase-2 (Cox-2) were assessed by Western blot, NF-kB activity and prostaglandin E2 (PGE2) were tested by ELISA. Exposure to Acla (even at 0.25 µg/ml) alone or SN alone (20 ng/ml) for 20 h did not induce significant apoptosis. However, SN (5 to 20 ng/ml) promoted apoptosis induced by 0.1 µg/ml Acla in a dose-dependent manner. Increased Caspase 3 and 9 protein expression correlated positively with SN concentration when combined with 0.1 µg/ml Acla. In addition, SN significantly inhibited Acla-induced NF-kB activation, Cox-2 gene expression and PGE2 production. In summary, SN most likely enhances apoptosis by suppressing NF-kB activation, and Cox-2 and PGE2 expression. In the future, this natural agent may provide a new avenue for anti-leukemia treatment.
 
Article
Radiation can induce the acute and chronic radiation sickness via physical, chemical and biological mechanisms. In this study, we further investigated the therapeutic effects of gingerol on radiated mice. Mice were given gingerol at 800 mg/kg B.W. by oral gavage once daily for 5 consecutive days after exposure to 1, 3, 5 and 7 Gy of 60 Co-gamma-radiation. Gingerol treated mice after irradiated by 3 and 5 Gy significantly increased the spleen index compared to 3 and 5 Gy irradiation+DDW mice, respectively. Besides, the gingerol treatment facilitated the recovery of white blood cells (WBC), lymphocytes (LYM), red blood cell (RBC) and hemoglobin (HGB) cell number at all the exposure doses. Moreover, the irradiated mice showed a dose-dependent depletion in superoxide dismutase (SOD) activities and total anti-oxidative capacity (T-AOC), while elevation in the malondialdehyde (MDA) contents, however, gingerol treated mice after irradiation increased activities of SOD and T-AOC, and decreased the MDA levels in liver to a certain extent in all the exposure dose groups. In addition, the gingerol reduced the numbers of micronuclei (MN) of polychromatic erythrocytes (PCEs) in the bone marrow after mice were irradiated by 0, 1, 3, 5 Gy of γ-ray. Together, the findings indicated that gingerol had the therapeutic effects on mice against hematopoietic suppression and anti-oxidative damage caused by irradiation.
 
Article
This study determined the effect of 30% ethanol extract of plant mixture (EPM: Glycyrrhiza uralensis, Angelica gigas, Acorus calamus, Cnidium officinale, Panax ginseng, Camellia sinensis, Salvia miltiorrhiza, Zanthoxylum schinifolium, Carthamus tinctorius, Prunus persica and Scrophularia buergeriana) on promotion of hair growth in human hair dermal papilla cells and C57BL/6J mice. EPM significantly increased the proliferation of human hair dermal papilla cells in a dose-and time-dependent manner (p < 0.05 and p < 0.01). EPM also enhanced mRNA and protein levels of growth factors such as IGF-1, VEGF, KGF and HGF. Moreover, photographical and histological observation showed that application of EPM resulted in the early onset and prolongation of the anagen phase of the hair cycle. In addition, the EPM was revealed to possess potent inhibitory effect on the 5α-reductase activity. Taken together, these results suggest that EPM has hair growth promoting potential and can be used for hair growing products.
 
Estimation of IC50 of A. sativum in L.major (MRHO/IR/75/ER) promastigotes. Promastigotes (1x10 6 /250 µl/well) were incubated with concentrations of A. sativum (0-100 µg/ml) for 72 h and viability was measured. Each point corresponds to the mean ± SD of at least three experiments in triplicates.
Morphology of L. major (MRHO/IR/75/ER) promastigotes after various time period’s treatment by A. sativum extract (37 μg/ml) in light microscopy (magnification, x 100). A top: Control group 48 h after sub culture and (A down) 48 h after A. sativum extract (37 μg/ml) treatment with. (B and C) Flow cytometry analysis of promastigotes after labeling with annexin-V FLUOS and PI in control group (B) and treated cells after 48 h (C). Miltefosine failed to make necrosis in L. major (MRHO/IR/75/ER) promastigotes even prolonged administration as all the cells remained negative for PI. Lower right region (LR) belongs to apoptotic cells (annexin positive) and upper left region (UL) belongs to necrotic cells (PI positive). (B) Control and (C) test sample 48 h after treatment. 
(A) The percentages DNA content of L. major (MRHO/IR/75/ER) promastigotes by flow cytometry, found in the sub-G1 peak: untreated (top) or treated with 37 μg/ml A. sativum (down). FL3-H, fluorescence intensity. B): DNA fragmentation analysis by agarose gel electrophoresis. The DNA of untreated (Lane 1), 37 μg/ml garlic extract-treated L. major (MRHO/IR/75/ER) promastigotes after 6 h and 12 h (Lanes 2 and 3) and A. sativum extract-treated L. major (MRHO/IR/75/ER) promastigotes after 24 h (Lane 4). 
(A, B): Relative gene expression of L.major prmastigtes (A) Metacaspase gene expression in treated and control group. (B) PARP gene expression in treated and control group after different time points.The bars show mean value of PARP and metacaspase gene expression in each group. Molecular analyst software was used to analysis of density of each band of PCR products. The results indicate intensity of expression for each gene after different time points. PARP and metacaspase gene expression responses to A. sativum extract (37 µg/ml) were also expressed as the expression index defined as the ratio. Results are shown relative to HPRT, defined as 100%. The values (mean± SEM) are derived from triplicate independent tests.
Article
Leishmaniasis is a protozoan disease that affects many people in the world, hence researchers are trying to design more effective and safer medicine, because of many limits in present chemical drug. In this regard, botany- derived products are the most attractive materials. The aim of this study was to determine the possible Allium sativum - induced cell death in Leishmania major (L. major) promastigotes. RPMI1640 cultured L. major promastigotes were subjected to different concentrations of A. sativum and viability of the parasites was assayed by MTT. Annexin V- FLUOS staining was performed to study apoptotic properties of the extract using FACS flow cytometry. Furthermore, metacaspase and PARP gene expression of treated L. major was studied. Apoptotic dose-dependent death of L. major accompanied by DNA fragmentation, cell contraction and externalization of phosphatidylserine with intact of integrity of plasma membrane was observed. In addition, overexpression of metacaspase and PARP was seen 4 h after treatment. Current study revealed that A. sativum extract induced apoptotic phenomenon in standard strain of L. major.
 
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Ali Ghasemzadeh
  • Universiti Putra Malaysia
Btc Cars IGKV Bilaspur
  • BTC College of Agriculture and Research Station (IGKV), Bilaspur (Chhattisgarh)
Kuntal Das
R. K. S. Tiwari
  • Indira Gandhi Agricultural University
Divya Shrivastava
  • Otago Polytechnic