Paeoniflorin Promotes Angiogenesis in A Vascular Insufficiency Model of Zebrafish in vivo and in Human Umbilical Vein Endothelial Cells in vitro

Objective: To investigate the pro-angiogenic effects of paeoniflorin (PF) in a vascular insufficiency model of zebrafish and in human umbilical vein endothelial cells (HUVECs). Methods: In vivo, the pro-angiogenic effects of PF were tested in a vascular insufficiency model in the Tg(fli-1:EGFP)y1 transgenic zebrafish. The 24 h post fertilization (hpf) embryos were pretreated with vascular endothelial growth factor (VEGF) receptor tyrosine kinase inhibitor Ⅱ (VRI) for 3 h to establish the vascular insufficiency model and then post-treated with PF for 24 h. The formation of intersegmental vessels (ISVs) was observed with a fluorescence microscope. The mRNA expression of fms-like tyrosine kinase-1 (flt-1), kinase insert domain receptor (kdr), kinase insert domain receptor like (kdrl) and von Willebrand factor (vWF) were analyzed by real-time polymerase chain reaction (PCR). In vitro, the pro-angiogenic effects of PF were observed in HUVECs in which cell proliferation, migration and tube formation were assessed. Results: PF (6.25–100 μmol/L) could rescue VRI-induced blood vessel loss in zebrafish and PF (25–100 μmol/L) , thereby restoring the mRNA expressions of flt-1, kdr, kdrl and vWF, which were down-regulated by VRI treatment. In addition, PF (0.001–0.03 μmol/L) could promote the proliferation of HUVECs while PF stimulated HUVECs migration at 1.0–10 μmol/L and tube formation at 0.3 μmol/L. Conclusion: PF could promote angiogenesis in a vascular insufficiency model of zebrafish in vivo and in HUVECs in vitro.     

Anti-Proliferative Effects of Ginsenosides Extracted from Mountain Ginseng on Lung Cancer

Objective

To investigate the effect of three major ginsenosides from mountain ginseng as anticancer substance and explore the underlying mechanism involved in lung cancer.

Methods

The inhibitory proliferation of lung cancer by major five ginsenosides (Rb1, Rb2, Rg1, Rc, and Re) was examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Calculated 50% inhibition (IC₅₀) values of five ginsenosides were determined and compared each other. Apoptosis by the treatment of single ginsenoside was performed by fluorescence-assisted cytometric spectroscopy. The alterations of apoptosis-related proteins were evaluated by Western blot analysis.

Results

The abundance of ginsenosides in butanol extract of mountain ginseng (BX-MG) was revealed in the order of Rb1, Rg1, Re, Rc and Rb2. Among them, Rb1 was the most effective to lung cancer cell, followed by Rb2 and Rg1 on the basis of relative IC₅₀ values of IMR90 versus A549 cell. The alterations of apoptotic proteins were confirmed in lung cancer A549 cells according to the administration of Rb1, Rb2 and Rg1. The expression levels of caspase-3 and caspase-8 were increased upon the treatment of three ginsenosides, however, the levels of caspase-9 and anti-apoptotic protein Bax were not changed.

Conclusion

Major ginsenosides such as Rb1, Rb2 and Rg1 comprising BX-MG induced apoptosis in lung cancer cells via extrinsic apoptotic pathway rather than intrinsic mitochondrial pathway.

     

Jinmaitong(筋脉通)Alleviates the Diabetic Peripheral Neuropathy by Inducing Autophagy

Objective:To observe the deregulation of autophagy in diabetic peripheral neuropathy(DPN) and investigate whether Jinmaitong(筋脉通i,JMT) alleviates DPN by inducing autophagy.Methods:DPN models were established by streptozotocin-induced diabetic rats and Schwann cells(SCs) cultured in high glucose medium.The pathological morphology was observed by the improved Bielschowsky's nerve fiber axonal staining and the Luxol fast blue-neutral red myelin staining.The ultrastructure was observed by the transmission electron microscopy.Beclinl level was detected by immunohistochemistry and Western blot.The proliferation of cultured SCs was detected by methylthiazolyldiphenyl-tetrazolium bromide.Results:Diabetic peripheral nerve tissues demonstrated pathological morphology and reduced autophagic structure,accompanied with down-regulation of Beclinl.JMT apparently alleviated the pathological morphology change and increased the autophagy[in vivo,Beclinl integral optical density(IOD) value of the control group 86.6±17.7,DM 43.9±8.8,JMT 73.3 ±17.8,P0.01 or P0.05,in vitro Beclinl IOD value of the glucose group 0.4710.25 vs the control group 0.88 ±0.29,P0.05].Consequently,inhibition of autophagy by 3-methyladenine resulted in a time- and concentrationdependent decrease of the proliferation of SCs(P0.05,P0.01).Conclusions:Down-regulation of autophagy in SCs might contribute to the pathogenesis of DPN.JMT alleviates diabetic peripheral nerve injury at least in part by inducing autophagy.     

Effects of Senegenin against Hypoxia/Reoxygenation-Induced Injury in PC12 Cells

Objective:To investigate the effect and the potential mechanism of Senegenin(Sen) against injury induced by hypoxia/reoxygenation(H/R) in highly differentiated PC12 cells.Methods:The cultured PC12 cells were treated with H/R in the presence or absence of Sen(60 μmol/L).Four groups were included in the experiment:control group,H/R group,H/R+Sen group and Sen group.Cell viability of each group and the level of lactate dehydrogenase(LDH) in culture medium were detected for the pharmacological effect of Sen.Hoechst 33258 staining and annexin V/propidium iodide double staining were used to analyze the apoptosis rate.Moreover,mitochondrial membrane potential(△Ψm),reactive oxygen species(ROS) and intracellular free calcium([Ca~(2+)]i) were measured by fluorescent staining and flow cytometry.Cleaved caspase-3and activity of NADPH oxidase(NOX) were determined by colorimetric protease assay and enzyme linked immunosorbent assay,respectively.Results:Sen significantly elevated cell viability(P0.05),decreased the leakage of LDH(P0.05) and apoptosis rate(P0.05) in H/R-injured PC12 cells.Sen maintained the value of△Ψm(P0.05) and suppressed the activity of caspase-3(P0.05).Moreover,Sen reduced ROS accumulation(P0.05) and[Ca~(2+)]i increment(P0.05) by inhibiting the activity of NOX(P0.05).Conclusion:Sen may exert cytoprotection against H/R injury by decreasing the levels of intracellular ROS and[Ca~(2+)]_i,thereby suppressing the mitochondrial pathway of cellular apoptosis.     

Pure Total Flavonoids from Citrus paradisi Macfad Induce Leukemia Cell Apoptosis In Vitro

Objective

To investigate the potential effect of pure total flavonoids from Citrus paradisi Macfad peel (PTFC) on the proliferation of human myeloid leukemia cells Kasumi-1, HL-60 and K562, and the underlying mechanisms.

Methods

PTFC was extracted from Citrus paradisi Macfad peel and was identified by high performance liquid chromatography. The effect of PTFC on the proliferation and apoptosis of leukemia cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, fluorescent microscopy and flow cytometry, respectively. The effect of PTFC on the expression levels of apoptosis-related regulators was determined by Western blot assay.

Results

Treatment with PTFC inhibited leukemia cell proliferation in a dose- and time-dependent manner and triggered Kasumi-1 cell apoptosis. Treatment with PTFC significantly increased the levels of activated poly adenosine diphosphate-ribosepolymerase and caspase-3/-9, but reduced the levels of Mcl-1 expression in Kasumi-1 cells. However, PTFC did not obviously induce HL-60 cell apoptosis.

Conclusion

PTFC inhibited leukemia cell proliferation and induced their apoptosis by modulating apoptosisrelated regulator expression in leukemia cells in vitro.

     

Salvianolate Reduces Murine Myocardial Ischemia and Reperfusion Injury via ERK1/2 Signaling Pathways in vivo

Objective

To analyze the effects of salvianolate on myocardial infarction in a murine in vivo model of ischemia and reperfusion (I/R) injury.

Methods

Myocardial I/R injury model was constructed in mice by 30 min of coronary occlusion followed by 24 h of reperfusion and pretreated with salvianolate 30 min before I/R (SAL group). The SAL group was compared with SHAM (no I/R and no salvianolate), I/R (no salvianolate), and ischemia preconditioning (IPC) groups. Furthermore, an ERK1/2 inhibitor PD98059 (1 mg/kg), and a phosphatidylinositol-3-kinase (PI3-K) inhibitor, LY294002 (7.5 mg/kg), were administered intraperitoneal injection (i.p) for 30 min prior to salvianolate, followed by I/R surgery in LY and PD groups. By using a double staining method, the ratio of the infarct size (IS) to left ventricle (LV) and of risk region (RR) to LV were compared among the groups. Correlations between IS and RR were analyzed. Western-blot was used to detect the extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (AKT) phosphorylation changes.

Results

There were no significant differences between RR to LV ratio among the SHAM, I/R, IPC and SAL groups (P>0.05). The SAL and IPC groups had IS of 26.1%±1.4% and 22.3%±2.9% of RR, respectively, both of which were significantly smaller than the I/R group (38.5%±2.9% of RR, P<0.05, P<0.01, respectively). Moreover, the phosphorylation of ERK1/2 was increased in SAL group (P<0.05), while AKT had no significant change. LY294002 further reduced IS, whereas the protective role of salvianolate could be attenuated by PD98059, which increased the IS. Additionally, the IS was not linearly related to the RR (r=0.23, 0.45, 0.62, 0.17, and 0.52 in the SHAM, I/R, SAL, LY and PD groups, respectively).

Conclusion

Salvianolate could reduce myocardial I/R injury in mice in vivo, which involves an ERK1/2 pathway, but not a PI3-K signaling pathway.

     

Aqueous Extracts of Tribulus terrestris Protects Against Oxidized Low-Density Lipoprotein-Induced Endothelial Dysfunction

Objective:To investigate the role of aqueous extracts of Tribulus terrestris(TT) against oxidized low-density lipoprotein(ox-LDL)-induced human umbilical vein endothelial cells(HUVECs) dysfunction in vitro.Methods:HUVECs were pre-incubated for 60 min with TT(30 and 3 μg/mL respectively) or 10~(-5) mol/L valsartan(as positive controls) and then the injured endothelium model was established by applying 100 μg/mL ox-LDL for 24 h.Cell viability of HUVECs was observed by real-time cell electronic sensing assay and apoptosis rate by Annexin V/PI staining.The cell migration assay was performed with a transwell insert system.Cytoskeleton remodeling was observed by immunofluorescence assay.The content of endothelial nitric oxide synthase(eNOS) was measured by enzyme-linked immunosorbent assay.Intracellular reactive oxygen species(ROS) generation was assessed by immunofluorescence and flow cytometer.Key genes associated with the metabolism of ox-LDL were chosen for quantitative real-time polymerase chain reaction to explore the possible mechanism of TT against oxidized LDL-induced endothelial dysfunction.Results:TT suppressed ox-LDL-induced HUVEC proliferation and apoptosis rates significantly(41.1%and 43.5%after treatment for 3 and 38 h,respectively;P0.05).It also prolonged the HUVEC survival time and postponed the cell's decaying stage(from the 69 th h to over 100 h).According to the immunofluorescence and transwell insert system assay,TT improved the endothelial cytoskeletal network,and vinculin expression and increased cell migration.Additionally,TT regulated of the synthesis of endothelial nitric oxide synthase and generation of intracellular reactive oxygen species(P0.05).Both 30 and 3 μg/mL TT demonstrated similar efficacy to valsartan.TT normalized the increased mRNA expression of PI3Kα and Socs3.It also decreased mRNA expression of Akt1,AMPKα1,JAK2,LepR and STAT3 induced by ox-LDL.The most notable changes were JAK2,LepR,PI3 K α,Socs3 and STAT3.Conclusions:TT demonstrated potential lowering lipid benefits,anti-hypertension and endothelial protective effects.It also suggested that the JAK2/STAT3 and/or PI3K/AKT pathway might be a very important pathway which was involved in the pharmacological mechanism of TT as the vascular protective agent.     

Qualitative Analysis of A Sulfur-fumigated Chinese Herbal Medicine by Comprehensive Two-Dimensional Gas Chromatography and High-Resolution Time of Flight Mass Spectrometry Using Colorized Fuzzy Difference Data Processing

Objective: To investigate the chemical transformation of volatile compounds in sulfur-fumigated Radix Angelicae Sinensis. Methods: A comprehensive two-dimensional gas chromatography(GC×GC) and high-resolution time-of-flight mass spectrometry(HR-TOF/MS) with colorized fuzzy difference(CFD) method was used to investigate the effect of sulfur-fumigation on the volatile components from Radix Angelicae Sinensis. Results: Twenty-five compounds that were found in sun-dried samples disappeared in sulfur-fumigated samples. Seventeen volatile components including two sulfur-containing compounds were newly generated for the first time in volatile oils of sulfur-fumigated Radix Angelicae Sinensis. Conclusion: The strategy can be successfully applied to rapidly and holistically discriminate sun-dried and sulfur-fumigated Radix Angelicae Sinensis. GC×GC-HR-TOF/MS based CFD is a powerful and feasible approach for the global quality evaluation of Radix Angelicae Sinensis as well as other herbal medicines.     

Brucine Inhibits Bone Metastasis of Breast Cancer Cells by Suppressing Jagged1/Notch1 Signaling Pathways

Objective: To examine the effects of brucine on the invasion, migration and bone resorption of receptor activator of nuclear factor-kappa B ligand(RANKL)-induced osteoclastogenesis. Methods: The osteoclastogenesis model was builded by co-culturing human breast tumor MDA-MB-231 and mouse RAW264.7 macrophages cells. RANKL(50 ng/m L) and macrophage-colony stimulating factor(50 ng/m L) were added to this system, followed by treatment with brucine(0.02, 0.04 and 0.08 mmol/L), or 10 μmol/L zoledronic acid as positive control. The migration and bone resorption were measured by transwell assay and in vitro bone resorption assay. The protein expressions of Jagged1 and Notch1 were investigated by Western blot. The expressions of transforming growth factor-β1(TGF-β1), nuclear factor-kappa B(NF-κB) and Hes1 were determined by enzyme-linked immunosorbent assay. Results: Compared with the model group, brucine led to a dose-dependent decrease on migration of MDA-MB-231 cells, inhibited RANKL-induced osteoclastogenesis and bone resorption of RAW264.7 cells(P 0.01). Furthermore, brucine decreased the protein levels of Jagged1 and Notch1 in MDA-MB-231 cells and RAW264.7 cells co-cultured system as well as the expressions of TGF-β1, NF-κB and Hes1(P0.05 or P0.01). Conclusion: Brucine may inhibit osteoclastogenesis by suppressing Jagged1/Notch1 signaling pathways.     

Relationship between Two Blood Stasis Syndromes and Inflammatory Factors in Patients with Acute Coronary Syndrome

Objective

To investigate the relationship between inflammatory factors and two Chinese medicine (CM) syndrome types of qi stagnation and blood stasis (QSBS) and qi deficiency and blood stasis (QDBS) in patients with acute coronary syndrome (ACS).

Methods

Sixty subjects with ACS, whose pathogenesis changes belongs to qi disturbance blood stasis syndrome, were divided into 2 groups: 30 in the QSBS group and 30 in the QDBS group. The comparative analysis on them was carried out through comparing general information, coronary angiography and inflammatory factors including intracellular adhesion molecule-1 (ICAM-1), chitinase-3-like protein 1 (YKL-40) and lipoprotein-associated phospholipase A2 (Lp-PLA2).

Results

Compared with the QSBS group, Lp-PLA2 and YKL-40 levels in the QDBS group showed no-significant difference (P>0.05); ICAM-1 was significantly higher in the QDBS group than in the QSBS group in the pathological processes of qi disturbance and blood stasis syndrome of ACS (P<0.05).

Conclusions

Inflammatory factor ICAM-1 may be an objective basis for syndrome typing of QSBS and QDBS, which provides a research direction for standardization research of CM syndrome types.

     

Curcumin Reduces Cardiac Fibrosis by Inhibiting Myofibroblast Differentiation and Decreasing Transforming Growth Factor β1 and Matrix Metalloproteinase 9 / Tissue Inhibitor of Metalloproteinase 1

Objective: To study the effect of curcumin on fibroblasts in rats with cardiac fibrosis. Methods: The rats were randomly divided into 4 groups(n=12 in each group): the normal control, isoproterenol(ISO), ISO combined with low-dose curcumin(ISO+Cur-L), and ISO combined with high-dose curcumin(ISO+Cur-H) groups. ISO+Cur-L and ISO+Cur-H groups were treated with curcumin(150 or 300 mg·kg-1·day-1) for 28 days. The primary culture of rat cardiac fibroblast was processed by trypsin digestion method in vitro. The 3rd to 5th generation were used for experiment. Western blot method was used to test the expression of collagen type Ⅰ/Ⅲ, α-smooth muscle actin(α-SMA), transforming growth factor(TGF)-β1, matrix metalloproteinase(MMP)-9 and tissue inhibitor of metalloproteinase(TIMP)-1. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetr-azolium bromide(MTT) assay was applied to test the proliferation of fibroblast. Result: Curcumin significantly decreased interstitial and perivascular myocardial collagen deposition and cardiac weight index with reducing protein expression of collagen type Ⅰ/Ⅲ in hearts(P0.05). In addition, curcumin directly inhibited angiotensin(Ang) Ⅱ-induced fibroblast proliferation and collagen type Ⅰ/Ⅲ expression in cardiac fibroblasts(P0.05). Curcumin also inhibited fibrosis by inhibiting myofibroblast differentiation, decreased TGF-β1, MMP-9 and TIMP-1 expression(P0.05) but had no effects on Smad3 in Ang Ⅱ incubated cardiac fibroblasts. Conclusions: Curcumin reduces cardiac fibrosis in rats and Ang Ⅱ-induced fibroblast proliferation by inhibiting myofibroblast differentiation, decreasing collagen synthesis and accelerating collagen degradation through reduction of TGF-β1, MMPs/TIMPs. The present findings also provided novel insights into the role of curcumin as an anti-fibrotic agent for the treatment of cardiac fibrosis.     

Protective Effects of Danlou Tablet (丹蒌片) against Murine Myocardial Ischemia and Reperfusion Injury In Vivo

Objective: To observe the in vivo effect of Danlou Tablet (丹蒌片, DLT) on myocardial ischemia and reperfusion (I/R) injury. Methods: DLT effects were evaluated in mouse heart preparation using 30-min coronary occlusion followed by 24-h reperfusion and compared among sham group (n=6), I/R group (n=8), IPC group (ischemia preconditioning, n=6) and DLT group (I/R with DLT pretreatment for 3 days, 750 mg?kg-1?day-1, n=8). The effects of DLT were characterized in infarction size (IS) compared with risk region (RR) and left ventricle using the Evans blue/triphenyltetrazolium chloride double dye staining method in vivo. Furthermore, the dose-dependent effect of DLT on I/R injury was evaluated by double staining method. Five different concentrations of DLT (0.625, 1.25, 2.5, 5 and 10 g?kg-1?day-1) were chosen in this study, and dose-response curve of DLT was obtained on these data. Results: The ratio of IS to left ventricle was significantly smaller in the DLT and IPC groups than the I/R group (P<0.05 or P<0.01), the ratio of IS to RR was also reduced in the DLT and IPC groups (P<0.01), while there were no differences in RR among the four groups (P>0.05). Experiments showed incidence of arrhythmias was reduced in the DLT group (P<0.01). Furthermore, DLT produced a dose-dependent inhibitory effect with a half maximal inhibitory concentration of 1.225 g?kg-1?day-1. Conclusions: Our research concluded that DLT was effective in reducing I/R injury in mice, and provided experimental supports for the clinical use of DLT.     

Effects of Modified Qing'e Pill (加味青娥丸) on Expression of Adiponectin, Bone Morphogenetic Protein 2 and Coagulation-Related Factors in Patients with Nontraumatic Osteonecrosis of Femoral Head

Objective: To observe the regulation of Chinese herbal medicine, Modified Qing'e Pill(加味青娥丸, MQEP), on the expression of adiponectin, bone morphogenetic protein 2(BMP2), osteoprotegerin(OPG) and other potentially relevant risk factors in patients with nontraumatic osteonecrosis of the femoral head(ONFH). Methods: A total of 96 patients with nontraumatic ONFH were unequal randomly divided into treatment group(60 cases) and control group(36 cases). The treatment group were treated with MQEP while the control group were treated with simulated pills. Both groups were given caltrate D. Six months were taken as a treatment course. Patients were followed up every 2 months. The levels of plasma adiponectin, BMP2, OPG, von Willebrand factor(vWF), von Willebrand factor cleaving protease(vWF-cp), plasminogen activator inhibitor 1(PAI-1), tissue plasminogen activator(tPA), C-reactive protein(CRP), blood rheology, bone mineral density(BMD) of the femoral head and Harris Hip Score were measured before and after treatment. Results: After 6 months of treatment, compared with the control group, patients in the treatment group had significantly higher adiponectin and BMP2 levels(P0.01 and P=0.013, respectively), lower vWF, PAI-1 and CRP levels(P=0.019, P0.01 and P0.01, respectively), and lower blood rheology parameters. BMD of the femoral neck, triangle area and Harris Hip Score in the treatment group were significantly higher than those in the control group. Moreover, plasma adiponectin showed a positive association with BMP2(r=0.231, P=0.003) and a negative association with PAI-1(r=–0.159, P0.05). Conclusions: MQEP may play a protective role against nontraumatic ONFH by increasing the expression of adiponectin, regulating bone metabolism and improving the hypercoagulation state, which may provide an experimental base for its clinical effects.     

Effect of Health Education Based on Integrative Therapy of Chinese and Western Medicine for Adult Patients with Type 2 Diabetes Mellitus: A Randomized Controlled Study

Objective: To investigate the effects of health education based on integrative therapy of Chinese and Western medicine for type 2 diabetes mellitus(T2DM) from the aspects of knowledge, attitude and practice(KAP), health-related quality of life(HRQo L), body mass index(BMI) and glucose control. Methods: Patients were individually randomized into intervention group(receiving integrative education, n=120) and control group(receiving usual education, n=120). The primary outcome was the changes in glycosylated hemoglobin A1c(HbA1c) levels after 3, 6, 9 and 12 months from baseline. Hierarchical linear models(HLMs) were used to assess within-group changes in outcomes over time and between-group differences in patterns of change. Secondary outcomes were KAP scores, HRQo L scores and BMI after 6 and 12 months, paired-sample t test was used to assess within-group changes in outcomes in 6 and 12 months, independent-sample t test was used to assess between-group differences in patterns of change. Results: HbA1c decreased statistically from baseline to 3 months, from 3 to 6 months, from 6 to 9 months and from 9 to 12 months in the intervention group(all P0.01); and decreased significantly from baseline to 3 months, and from 3 to 6 months in the control group(P0.01). There were significant between-group differences from baseline to 3 months(P=0.044), from 6 to 9 months(P0.01) and from 9 to 12 months(P0.01). Significant improvements in the intervention group along with significant between-group differences were found in KAP and HRQo L scores respectively(all P0.05). The number in the intervention group of normal weight increased from 56 at baseline to 81(6 months), 94(12 months), the number in the control group were 63(baseline), 69(6 months), 70(12 months), the χ~2 of hierarchical analysis of BMI were 6.93(P=0.075), 10.31(P=0.016), 15.53(P0.01), respectively. Conclusion: Health education based on integrative therapy of Chinese and Western medicine is beneficial to the control of T2DM and should be recommended for T2DM.     

Contribution of Water and Lipid Soluble Substances in the Relaxant Effects of Tymus Vulgaris Extract on Guinea Pig Tracheal Smooth Muscle in Vitro

Objective:To examine the relaxant effects of hydro-ethanolic,macerated aqueous(MA) and lipidfree macerated aqueous(LFMA) extract of Tymus vulgaris on tracheal chains of guinea pigs.Methods:The relaxant effects of five cumulative concentrations of each extract(0.4,0.8,1.2,1.6 and 2.0 g/100 mL) were compared with saline as negative control and five cumulative concentrations of theophylline(0.1,0.2,0.4,0.6 and 0.8 mmol/L) on precontracted tracheal smooth muscle of guinea pig with 60 mmol/L KCI(group 1) and 10 μmol/L methacholine(group 2,n=6 for each group).Results:In group 1 all concentrations of theophylline,three higher concentrations of hydro-ethanolic,two concentrations of LFMA and last concentration of MA extracts showed significant relaxant effects compared with that of saline(P0.05 or P0.01).Two lower concentrations of LFMA and all concentrations of MA except higher one caused contraction compared with saline(P0.05 or 0.01).In group 2 experiments,all concentrations of theophylline,hydro-ethanolic,MA and LFMA extracts showed significant relaxant effects compared to that of saline(P0.05 or P0.01).In both groups,the relaxant effect of all concentrations of hydro-ethanolic extract were significantly higher than most concentrations of others(P0.05 or P0.01).The relaxant effect of different concentrations of three extracts were significantly greater in group 2 compared with group 1 experiments(all P0.01).There were significantly positive correlations between the relaxant effects and concentrations for theophylline and ail extracts in both groups(P0.05 or P0.01).Conclusion:Hydro-ethanolic extract has a potent weaker relaxant effect for other extracts from Tymus vulgaris on tracheal chains of guinea pigs.     

In Vitro Antifungal Activity of Some Traditional Persian Medicinal Plants on Pathogenic Fungi

Objective

To investigate the antifungal activities of the extracts and sub-fractions of Phlomis olivieri, Verbascum speciosum, Sambucus ebulus and Erigeron hyrcanicus, four Persian medicinal plants used in Iranian folk medicine.

Methods

Evaluation of the antifungal activity was performed on the clinical isolates of pathogenic fungi including Aspergillus fumigatus, A. flavus, Trichophyton mentagrophytes, T. rubrum, T. verrucosum, Microsporum canis, M. gypseum and Epidermophyton floccosum, and the yeast Candida albicans. The susceptibility tests were done by agar well diffusion method. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of active extracts and sub-fractions were measured using method of National Committee for Clinical Laboratory Standards (NCCLS).

Results

Only P. olivieri sub-fractions were found to have fungicidal activity among the other investigated plants. The MIC and MFC was found to be high in petroleum ether, chloroform and ethyl acetate fractions (100 and 200 mg/mL) against the studied pathogenic fungi and the yeast Candida albicans. P. olivieri sub-fractions significantly inhibited the growth of all pathogenic fungi and the yeast studied.

Conclusion

If the antifungal activity of P. olivieri is confirmed by in vivo studies and if the responsible compound (s) is isolated and identified, it could be a good remedy for mycotic infections.

     

Effect of Miscanthus sinensis var. purpurascens Flower Extract on Proliferation and Molecular Regulation in Human Dermal Papilla Cells and Stressed C57BL/6 Mice

Objective: To investigate the hair growth-promoting effect of Miscanthus sinensis var. purpurascens (MSP) flower extracton on in vitro and in vivo models. Methods: MSP flower extract was extracted in 99.9% methanol and applied to examine the proliferation of human dermal papilla cells (hDPCs) in vitro at the dose of 3.92–62.50 μg/mL and hair growth of C57BL/6 mice in vivo at the dose of 1000 μg/mL. The expression of transforming growth factor β1 (TGF-β1), hepatocyte growth factor (HGF), β-catenin, substance P was measured by relative quantitative realtime polymerase chain reaction. Histopathological and immunohistochemical analysis were performed. Results: MSP (7.81 μg/mL) down-regulated TGF-β1 and up-regulated HGF and β-catenin in hDPCs (P<0.01). MSP (1000 μg/mL)-treated mice showed the earlier transition of hair follicles from the telogen to the anagen phase. The number of mast cells was lower in the MSP-treated mice than in other groups (P<0.05 vs. NCS group). Substance P and TGF-β1 were expressed in hair follicles and skin of the MSP group lower than that in negative control. Stem cell factor in hair follicles was up-regulated in the MSP-treated mice (P<0.01). Conclusion: The MSP flower extract may have hair growth-promotion activities.     

Antimicrobial Agents from Selected Medicinal Plants in Libya

Objective: To test the in vitro antimicrobial efficacy of water and methanol extracts of 23 plant species that are commonly used in Libyan folk medicine. Methods: The antimicrobial activity was determined using the well-diffusion method. Four test microorganisms were used namely, Escherichia coli , Salmonella species, Staphylococcus aureus and Bacillus subtilis . The minimum inhibitory concentration (MIC) was determined for the high biologically active crude plant extracts. Results: Among 23 medicinal plants used in the study, only 5 methanolic extracts [Rosmarinus offcinalis L., Carduus marianium L., Lantana camara L., Rhus tripartite (ueria) Grande, and Thymus capitatus (L.) Hoffm (link)] showed the highest antimicrobial activity against Staphylococcus aureus, Bacillus subtilis and Salmonella species, while 22 methanolic and aqueous extracts showed moderate to weak antimicrobial activity on all tested organisms. However 19 of the extracts showed no activity at all against Gram –ve and Gram +ve microorganisms. MIC was found to be 1.25 mg/mL (Thymus capitatus), 3 mg/mL (Rhus tripartite), 4 mg/mL (Carduus marianium), 5 mg/mL (Rosamarinus officinalis) and 5 mg/mL (Lantana camara), respectively. Conclusions: The present results revealed that, crude methanolic extracts of the investigated Libyan folk medicinal plants exhibited mild to high in vitro antibacterial activities against Gram-positive and Gram-negative microorganisms.