Showing papers in "Chinese Pharmacological Bulletin in 2009"

Experimental study of theacrine on antidepressant effects

Abstract: Aim To investigate the antidepressant effects of theacrine (TC).Methods The antidepressant effects of TC were examined by a series of experiments including tail suspension test,forced swimming test,ambulatory activity test,yohimbine induced toxicity test,reserpine test and 5-HTP induced head-twitching test(The doses of groups were 3,10,30 mg·kg-1,bid respectively for continued 7 days of intragastric administration).Results Compared with the control group,TC could effectively shorten the immobility time in the tail suspension test (P0.05)and enhance the rotatory locomotor activities during forced swimming test (P0.05),but didn't affect the ambulatory activity.Moreover,TC also enhanced the mortality induced by yohimbine,and increased the head-twitching number induced by 5-HTP (P0.01).In addition,TC could markedly improve such symptoms as hypothermia (P0.01),akinesia (P0.05),and eye ptosis of mice (P0.01) induced by reserpine.Conclusions TC has antidepressant effects in various depression models of mice,which may be contributed to its influence on monoamine neurotransmitter.

Pharmacokinetic study of saikosaponin a in rat plasma by LC-ESI-MS

Abstract: Aim To establish an LC-ESI-MS method for determination of saikosaponin a and investigate its application to pharmacokinetic study in rats.Methods 12 rats were given 5 or 50 mg·kg-1 saikosaponin a via iv or ig.Drug plasma concentration was determined by LC-ESI-MS and pharmacokinetic parameters were evaluated by DAS Software.Results Calibration curve was linear between 0.025~5 mg·L-1 and LOQ was 25 μg·L-1,the recoveries of saikosaponin a from plasma over than 80%,and RSD of inter-day and intra-day assay was limited in 10%.After iv administration of 5 mg·kg-1 saikosaponin a,the pharmacokinetic parameters of Tmax,Cmax,AUC0-t,T1 2β,CL and Vd were 5 min,1 907 μg·L-1,6 4370 mg·h-1·L-1,100.6 min,0.0867 L·min-1·kg-1,21.89 L·kg-1 respectively.Conclusion The above-mentioned method is sensitive and specific,and suitable for pharmacokinetic studies of saikosaponin a in rats.

The effect of Cistanche deserticola polysaccharides(CDPS) on marcrophages activation

Abstract: Aim To investigate the activation effect of Cistanche deserticola polysaccharides(CDPS) on marcrophages.Methods The phagocytic ability of peritoneal macrophage(MΦ) was evaluated with neutral red dye phagocytosis assay;nitric oxide(NO) production was examined by Griess reaction;the activity of tumor necrosis factor-α(TNF-α) and interleukin-1(IL-1) secreted were measured by L929 cells bioassay and proliferation of mouse thymocyte.Results CDPS at the concentration range of 6.25～50 mg·L-1 enhanced the phagocytic ability and NO release of peritoneal macrophage in a dose-dependent manner.CDPS at the dose of 2.8～100 mg·L-1 significantly increased NO secretion of normal or immunesuppressed RAW264.7 cells.TNF-α production by RAW264.7 cells was promoted after incubation of RAW264.7 cells with CDPS at the concentration of 0.56～7.2 mg·L-1.IL-1 release of RAW264.7 cells was obviously elevated by CDPS at the dose of 3.6～10 mg·L-1,and they both showed a good dose-effect relationship.Conclusions CDPS can markedly promote the phagocytic and secretory functions of MΦ,and activate MΦ,which suggests that the immunomodulatory effect of CDPS may be related to it.

Preliminary study on cardiotoxicity of celastrol to zebrafish embryo

Abstract: Aim To study the cardiotoxicity of celastrol to zebrafish embryo.Method 48 h post-fertilization zebrafish embryos were used as model for analysis of heart toxicity and were treated with various concentrations of celastrol.6,12,24 h after treatment morphological and functional changes of embryos hearts were observed.Results Cardiotoxicity was not found in embryos during 24 h treatment with 1 μmol·L-1 concentration of celastrol.Toxic symptoms of embryos were caused by 2,3,4 μmol·L-1 concentrations of celastrol with appearance of heart linearization,heart membrane hemorrhage and hemocytes accumulation in cardiac region.Moreover,heart rate decreased significantly with increase of concentration and prolongation of treatment.The EC50(24 h)of decrease of heart rate was about 1.78 μmol·L-1.Conclusion Celastrol is cardiotoxic to zebrafish embryo.

Protection and its mechanism of two flavone morphons from Yulangsan on hypoxia-reoxygenation induced injury in myocardial cells

Abstract: Aim To observe the protective effects of two flavone morphons on hypoxia/reoxygenation(H/R) injury in myocardial cells and explore the mechanisms.Methods The hypoxia/reoxygenation injury model of cultured neonatal rat cardiomyocytes was developed. The cell morphology,the spontaneous beating,the survival rate,the content of LDH,NOS,T-SOD,MDA and the activity of Na+,K+-ATP,Ca2+,Mg2+-ATP enzyme were determined in the cultured neonatal rat cardiomyocytes injuried by H/R.Results Compared with the model,preconditioning by two flavone morphons enhanced the spontaneous beating and the survival rate. Meanwhile,it increased the activity of T-SOD,Na+,K+-ATP,Ca2+,Mg2+-ATP enzyme and decreased the release of LDH,NOS,MDA(P0.05) in a dose-dependent manner.Conclusion The two flavone morphons have good effects on hypoxia/reoxygenation injury in cultured neonatal rat cardiomyocytes,which is related with scavenging free radicals,inhibiting Ca2+-overloading.

The effect of Jiaweisinisan on cAMP response element binding protein and phosphorylation in PC12 cells injured by Corticosterone and Glutamate

Abstract: Aim To study the effect of Chinese prescription-Jiaweisinisan(JWSNS)on cAMP response element binding protein(CREB) and phosphorylation in PC12 cells injured by Corticosterone(Cort) and Glutamate(Glu).Method 200 μmol·L-1 Cort and 50 μmol·L-1 Glu were used to injure PC12 cells which were taken as the pharmacological models in vitro.JWSNS drug-containing serum was prepared and immunohistochemical and Western blot methods were adopted to detect the expressions of CREB and p-CREB in PC12 cell models.Results 200 μmol·L-1 Cort and 50 μmol·L-1 Glu could decrease the expressions of CREB and p-CREB in PC12 cells significantly(the CREB positive cell rate was 17.1%±4.2% and 20.8%±5.7% respectively,the expression of p-CREB was 0.587±0.123 and 0.515±0.157 respectively)(P0.05 or P0.01),while 10% JWSNS drug-containing serum increased those significantly(the CREB positive cell rate was 62.6%±11.7% and 79.5%±7.6% respectively,the expression of p-CREB was 1.298±0.112 and 1.269±0.128 respectively)(P0.05 or P0.01).Conclusion JWSNS can protect nerve and anti-depression by regulating cAMP-CREB signal pathway.

Study on the molecular mechanism of C-phycocyanin from Spirulina platensis induced apoptosis in HeLa cells

Abstract: Aim To investigate the influence and molecular mechanism of C-phycocyanin(CPC) from Spirulina platensis on apoptosis of HeLa cells in vitro.Methods Firstly,the effect of purified CPC on proliferation of HeLa cells in vitro was determined by MTT assay,and then electron microscope was exploited to observe the characteristic apoptotic features of cells treated with CPC.Subsequently,genomic DNA changes of HeLa cells were observed by agarose electrophoresis.Flow cytometric analysis revealed the influence of different concentrations of CPC on cell cycle of HeLa cells.The expressions of apoptosis related genes of CPC treated HeLa cells were determined by immunohistochemistry analysis.In addition,the activities of caspases and the release of cytochrome c from mitochondria into the cytosol were detected.Results Compared with control cells untreated with CPC,a significant decreased in the numbers of HeLa cells in survival treated with CPC and concentration dose effects existed.CPC could induce characteristic apoptotic features including cell shrinkage,membrane blebbing,microvilli loss,chromatin margination and condensation into dense granules or blocks.DNA of HeLa cells treated with CPC showed fragmentation pattern(DNA ladder of oligomers of 180~200 bp) typical for apoptotic cells.HeLa cells treated with different concentrations of CPC demonstrated an increasing percentage of cells in sub-G0/G1 phase.In addition,CPC could promote the expression of pro-apoptotic gene(Fas and ICAM-1);meanwhile,held back the anti-apoptotic gene(Bcl-2) expression,and then facilitated the transduction of tumoral apoptosis signals that resulted in the apoptosis of HeLa cells in vitro.In CPC treated HeLa cells,CPC treatment of HeLa cells also resulted in activation of caspases and release of cytochrome C from mitochondria into the cytosol.Conclusion C-phycocyanin from Spirulina platensis can induce the apoptosis of HeLa cells in vitro.By virtue of the promotion of the apoptosis signals transduction in HeLa,CPC realizes its antitumor activities.

Arecoline improved glucose and lipid metabolism in type 2 diabetic rats

Abstract: Aim To investigate the effects of arecoline on glucose and lipid metabolism in type 2 diabetic rats and its mechanisms in glucose metabolism.Methods A type 2 diabetic rat model was established by fed with high fructose-high fat diet.The animals were randomly divided into 7 groups: control group,high fructose-high fat diet group(HF)and high fructose-high fat diet+arecoline(1 mg·kg-1,5 mg·kg-1,10 mg·kg-1,20 mg·kg-1,50 mg·kg-1)groups.The blood glucose,lipid level,hepatic function and liver histology were measured.The mRNA expression of liver glucose-6-phosphatase(G6Pase),phosphoenolpyruvate carboxykinase(PEPCK),Forkhead Box O1(FoxO1)and peroxisome proliferator-activated receptor-γ coactlvator-1α (PGC-1α)were observed through RT-PCR.Results In comparison with the high fructose-high fat diet group,the fasting blood glucose and TC of the rats were significantly decreased by arecoline in a dose-dependment manner in high fructose-high fat diet+arecoline group.But hepatic function was damaged by 10 mg·kg-1,20 mg·kg-1 and 50 mg·kg-1 arecoline.The mRNA expression of hepatic G6Pase,PEPCK,FoxO1 and PGC-1α was decreased by treatment with 1 mg·kg-1 and 5 mg·kg-1 arecoline compared with the high fructose-high fat diet group.Conclusions Low dose arecoline can decrease fasting blood glucose and TC in type 2 diabettic rats,and the mechanism in glucose metabolism may be related to its effect on the inhibition of hepatic gluconeogenesis.

Effects of biatractylolide on the AD rats induced by Aβ_(1-40)

Abstract: Aim To observe the effects of biatractylolide on ability of remembering and the content of CHE in AD rats induced by Aβ1-40 to study the mechanism of biatractylolide.Methods 48 rats were randomly divided into 6 groups:the control group,the model group,the positive control group and 3 experimental drug-treated groups.All groups except the control were injected Aβ1-40 into the lateral cerebral ventricle.After administration for 10 days,the ability of remembering of rats was studied by the water maze method and the content of CHE was measured by the method of colorimetry.Results The swimming time of 3 experimental groups was obviously shorter than that of model group and the mistakes in the maze were much fewer (P0.05 or P0.01).The content of CHE of 3 experimental groups was obviously lower than that of model group.Conclusion Biatractylolide can improve the studying memory of the AD rats induced by Aβ1-40 and reduce CHE activity.

Protective effects of fenazinel dihydrochloride on focal cerebral ischemic injury in rats

Abstract: Aim To investigate the protective effect of fenazinel dihydrochloride(FD) on focal cerebral ischemia injury in rats and to explore its possible mechanism.Methods Middle cerebral artery occlusion(MCAO) was used to induce focal cerebral ischemia model.After 24 h ischemia,the infarct area was determined by the method of TTC staining,and the ischemia brains of some other rats were used for measurements of super oxide dismutase(SOD) activity,malondialdehyde(MDA) content,lactic acid(LA) content and the histopathological change.Cortical neurons of fetal rats were cultured in vitro.The protective effects of FD on injuries by treating cells with FBS retraction medium and with N-methyl-D-aspartic acid(NMDA) were observed by determinations of cell death rate,activity of SOD and change of MAD/LA level.Results FD decreased the infract area in MCAO model,increased the activity of SOD in brain,lowed the MDA/LA level,and protected against the brain histological injury.In neuron cells,FD not only increased neuron survival rate and activity of SOD,but also downregulated the MDA/LA level.Conclusion FD is able to protect cerebral ischemia with involvement of decreasing NMDA level,attenuating oxygen-derived free radicals and improving energy metabolism.

Protective effects of timosaponin BII on primary neurons against beta amyloid peptide 25-35

Abstract: Aim To study the protective effects of timosaponin BⅡ on primary neurons against beta amyloid peptide 25-35 induced toxicity.Methods Immunofluorescence was used to identify primary neurons.MTT(tetrazolium salt) assay was used to measure neurons metabolic state.Spectrophotometric method was used to measure the release of LDH(lactate dehydrogenase),the activity of SOD(superoxide dismutase),the activity of AChE(acetylcholine) and the production of MDA(malonaldehyde) in the culture medium.Results Treatment with beta amyloid peptide 25-35(20 μmol·L-1) for 24 h caused a significant damage in primary neurons.Timosaponin BⅡ 10-4、10-5 mol·L-1 markedly improved neurons metabolic activity,decresed the release of LDH and the production of MDA,markedly increased the activity of SOD and decresed the activity of AChE.Conclusion Timosaponin BⅡ could significantly reduce the neurotoxicity induced by beta amyloid peptide 25-35 in primary neurons.The mechanism of which may be related with resisting oxidative damage and regulating the cholinergic system.

Progress in research of antitumor mechanisms of marine polysaccharides

Abstract: polysaccharide has become a research focus in tumor therapy because of possessing positive immunomodulating and antitumor activities.At present,a series of polysaccharides with novel structures and unique function have been isolated from marine resources,which are potential choices for anti-tumor drugs.However,few reviews are reported about the antitumor mechanism of marine polysaccharide,so the progress in research of antitumor mechanisms of marine polysaccharide are reviewed in this paper.

Proliferation-inhibited and apoptosis-inducted effects of evodiamine on human hepatoma cell line HepG2

Abstract: Aim To explore the proliferation-inhibited,apoptosis-induced and cell cycle-regulated effect of evodiamine on human hepatoma cell line HepG2.Methods MTT,Dapi assay,flow cytometry analysis,comet assay were used.Results Evodiamine could significantly inhibit the growth of human hepatoma cell line HepG2.After 72 hours of treatment with evodiamine at different concentrations(64,16,4,1,0.25 μmol·L-1),the inhibitory rate of HepG2 was 74.0%,69.0%,60.5%,44.0% and 16.4%,respectively.Meanwhile,HepG2 showed typical apoptosis.After 24 and 36 hours' treatment with evodiamine(1 μmol·L-1),a typical subdiploid peak before G0/G1 phase was observed by flow cytometry and cell cycle was arrested in the G2/M phase,while the rate of apoptosis was 4.4%,18.0% and 30.3% of treatment with evodiamine for 12,24 and 36 hours respectively.After 24 and 36 hours' treatment with evodiamine(1 μmol·L-1),the average optical density was lower than that of the control and the length of tail increased compared with the control.Meanwhile the changes were related with time.Conclusion Evodiamine inhibits the proliferation and induces apoptosis of HepG2.

Tibet-medicine effects on β-amyloid pathology in a transgenic mouse model of Alzheimer's disease

Abstract: Aim To investigate the effect of Ratanasampil(RNSP),a traditional Chinese Tibet medicine on the β-amyloid peptide as a therapeutic target in Alzheimer's disease.Methods 25 mice of 13～14 months old female Tg2576 and C57BL/6XDBA control mice were used in drug test.All mice sacrificed after 8 weeks of RNSP treatment at 15～16 months of age.Open field activity and Y-maze tests were performed for animal behavioral change and memory ability.The serum levels of β-amyloid(Aβ) peptides and beta-site amyloid precursor protein cleaving enzyme-1(BACE-1) were measured by Western blot.Immunostaining with 1E8(1:25) was carried out on brain sections of drug and vehicle-treated mice.Results Drug-treated Tg2576 mice decreased training time by Y-maze tests(P0.01).Two months RNSP treatment in Tg2576 mice caused a decrease in open field behavior.The level of Aβ and BACE-1 in the serum was significantly decreased after RNSP treatment(P0.01).Preliminary plaque counting of the sections showed reduced plaque numbers in the drug-treated mice in brain.Conclusions Ratanasampil can improve the learning and memory ability of Tg2576 mice.It may reduce beta-amyloid generation by directly inhibiting BACE-1 activity.

Studies on the anti-inflammatory molecular mechanism of chlorogenic acid extracted from Lonicera confusa DC in vitro

Abstract: Aim To investigate the anti-inflammatory molecular mechanism of chlorogenic acid extracted from Lonicera confusa DC in vitroMethods PMΦ of a rat was segregatedMTT assay was used to detect the effects of the chlorogenic acid on PMΦ cells growth activities PMΦ was stimulated with LPS for a prolonged period,ELISA was used to detect the level of TNF-α,IL-6 and PGE2 in the supernatant;COX-2 activity was determined by the level of PGE2 in the supernatantAfter stimulating PMΦ with A23187 for a short time,the 6-keto-PGF1α level in the supernatant was measured by radioimmunoassay to express COX-1 activityResults Chlorogenic acid had no inhibitive effects between 3125 mg·L-1 and 1000 mg·L-1The level of TNF-α,IL-6 and PGE2 in drug groups was lower than that of LPS-induced group,and the difference was significant,in a dose-dependent mannerThe concentration of 50 mg·L-1 group was ineffective in the expression of TNF-αLow concentration chlorogenic acid inhibited the expression of 6-keto-PGF1α,while high-concentration induced itConclusions The anti-inflammatory effect of chlorogenic acid may be related to inhibiting TNF-α,IL-6 activity and affecting exogenous AA metabolism

Induction of apoptosis by Tryptanthrin on K562 cells

Abstract: Aim To study the effect of Tryptanthrin(Try) on proliferation and apoptosis of erythroleukemia K562 cells.Methods The cell proliferation effect of Try(1.56～50 mg·L-1) on K562 cells was assessed by MTT assay.The morphologic change was observed by Hoechst 33258 fluore-scent stain.The flow cytometer was used to detect cell apoptosis and cell cycle.Results MTT showed that in the range of 3.12～50 mg·L-1 Try obviously inhibited the proliferation of K562 cells in a dose and time-dependent manner.Typical apoptosis changes were observed in K562 cells treated with Try for 48 h by flourescence inverted microscope.With Annexin V-FITC and PI double staining,folw cytometer result showed that the apoptosis state was obvious in K562 cells treated with 25,50 mg·L-1 Try for 48 h.The cell cycle distribution of K562 was changed.The G0/G1 phase was blocked and the DNA synthesis was inhibited,accompanied with subdiploid apoptotic peak.Conclusion Try has an effect on inhibiting the cell proliferation and inducing the apoptosis of K562.

The apoptosis-inducing effect of MnSOD_m on K562 cells and its molecular mechanism

Abstract: Aim To explore the apoptotic effect of mimics of manganese superoxide dismutase(MnSODm)on human leukemia cell line K562 in vitro and the possible molecular mechanisms.Methods Human leukemia K562 cells were used as the target cells.The cell proliferating activity was examined by a MTT colorimetric assay,and the apoptosis of K562 cells was assessed with FITC-Annexin V and propidium iodide(PI)double staining and morphological changes.The expressions of bcl-2 and bax mRNA were detected by reverse transcription polymerase chain reaction(RT-PCR),and flow cytometry(FCM)was employed to measure the expressions of Bcl-2 and Bax protein,mitochondrial inner membrane potential(Δψm),Cytochrome C(Cyt C)release and Caspase-3 activity.Results The proliferation of K562 cells was obviously inhibited by 0.5~10 mg·L-1 MnSODm(P0.01),and the apoptosis rate of K562 cells detected with Annexin V/PI staining was markedly increased,and the typical apoptotic morphological changes of K562 cells was observed under optical and electronic microscopic morphology.The down-regulation of expressions of bcl-2 mRNA and protein was accompanied with the up-regulation of bax mRNA and protein.The disruption of mitochondria Δψm and the increased release of Cyt C from mitochondria to cytoplasm were noted,and the activity of Caspase-3 was significantly enhanced.Conclusions MnSODm can induce apoptosis of K562 cells via mitochondrial pathway by regulating the expression of Bax/Bcl-2.

Research on evaluation methods of the fever syndrome model based on metabonomics

Abstract: Aim To establish a set of consummate evaluation methods of animal models which are with syndromes of traditional Chinese medicine.Method By means of 2,4-dinitrophenol-induced fever syndrome model for the entry point,making use of meta bonomics as the platform,through analyzing metabolic fingerprint data of rat urine in the control and model group and metabolome of rat urine in model group at different time intervals,to approach the evaluation methods of animal models.Results Through research by metabonomics,the results showed that 2,4-dinitrophenol-induced animal models consisted with clinical fever syndrome of traditional Chinese medicine.Conclusion Metabonomics can be used for evaluation studies of animal models which are with syndromes of traditional Chinese medicine.

Effect of artificial musk on myocardial ischemia in animals

Abstract: Aim To observe the effect of artificial musk on myocardial ischemia in animals.Methods The animals were divided into five groups randomly:model group,nitroglycerin control group(the rats were 0.083 mg·kg-1,the mice were 0.042 mg·kg-1),artificial musk low,middle and high groups (the rats were 5,10,20 mg·kg-1,the mice were 2.5,5.0,10 mg·kg-1). To mean survival time of hypoxia,the experimental myocardial ischemia was induced by injection of pituitrin iv in adult rats;to blockade the coronary artery and the diversity of electrocardiographic sign,myocardial enzymes,the myocardial infarct size and pathology were recorded and examined.Results The survival time in hypoxia (P0.01 or P0.05) was prolonged,the experimental myocardial ischemia by injection of pituitrin(P0.05) was ameliorated,the myocardial enzymes were inhibited and the myocardial infarct size(P0.05) was diminished by artificial musk.Conclusions Artificial musk has protective effects on myocardial ischemia in animals.

Protective effects of geniposide on human umbilical vein endothelial cell injury induced by H_2O_2 in vitro

Abstract: Aim To study the protective effect and mechanism of geniposide on human umbilical vein endothelial cell(HUVEC) injury induced by H2O2.Methods The injured model was established by HUVEC treated with H2O2.HUVECs were cultured in vitro and divided into five groups:control group,H2O2 group and geniposide with different concentrations plus H2O2 group respectively.HUVECs were incubated with 400 μmol·L-1 H2O2 for 12 hours in the absence or presence of various concentrations of geniposide pre-incubation.Survival rate of HUVECs was determined by tetrazolium assay.The intracellular activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),total nitric oxide synthase(NOS) and extracellular nitric oxide(NO) level were detected.The intracellular reactive oxygen species(ROS) level and the apoptotic index and cell cycle alteration were detected by flow cytometry.Results Geniposide concentration-dependently increased the viability of injured endotheial cells,while increased the activity of SOD,GSH-Px,NOS and NO production.The intracellular ROS level and the apoptotic index were reduced by geniposide.The cell proliferation was increased with geniposide incubation.Conclusion Geniposid may be a potential anti-oxidation agent which has a protective effect against HUVEC injuries induced by H2O2.

Transcytosis mechanism of chlorogenic acid across cell monolayer model

Abstract: Aim To investigate the transcytosis mechanism of chlorogenic acid(CGA)by using Caco-2 and MDCK(Madin Darby canine kidney) monolayers models.Method ① Caco-2 and MDCK cell models:Caco-2 cell(105 cells/cm2) and MDCK cell(5×104 cells/cm2) were inoculated in Millicell-CM culture plate inserts,and the TEER of cell monolayer were detected to make sure the models are available for experiments.② Permeating experiments: to measure the value of OD of CGA and calculate the cumulative amount.Result CGA could be Absorbed and secreted on two monolayer models.Verapamil could inhibit the secretion at lower concentration of CGA on MDCK monolayer model.P-pg could partly act on the secretion of CGA on Caco-2 and MDCK cell models.Conclusion CGA can secrete and Absorb at the same time across Caco-2 and MDCK cell monolayers,P-pg partly involving in the secretion of CGA.

Effects of astragalosides on the expression of BDNF,TrkB and p75NTR mRNA against focal cerebral ischemia-reperfusion injury

Abstract: Aim To observe the neurological protective effects of astragalosides(AST) on focal cerebral ischemia-reperfusion(I/R) injury in rats and to explore its possible mechanism.Methods Male SD rats received right middle cerebral artery occlusion for 120 min,and were decapitated 1,3,7,and 14 days after reperfusion.AST(40 mg·kg-1) was orally administered after I/R.Neurological deficit score was daily determined,the expressions of BDNF and p75NTR mRNA were detected by RT-PCR,and the expression of TrkB mRNA was detected by real-time PCR.Results AST reduced the neurological deficit score on days 3,increased the expression of BDNF mRNA on days 3,7 and 14,decreased p75NTR mRNA and increased TrkB mRNA on days 3 and 7.Conclusions AST improves the neurological deficits after I/R in rats.The mechanism may be related with increasing BDNF,and TrkB mRNA,and decreasing p75NTR mRNA.

Protective effect of extract of astragalus against injury induced by hypoxia/reoxygenation in hippocampus neuron

Abstract: Aim To study the effect of EA on the injury induced by hypoxia/reoxygenation in primary cultures of rat hippocampal neurons.Methods Rat hippocampal neurons in primary culture were used,and a apoptosis model was induced by hypoxia/reoxygenation.MTT assay and LDH releasing rate were used to detect the cell viability.The apoptosis rate of hippocampal neurons was analyzed by Hoechst 33258 staining,flow cytometry with AnnexinV-FITC and PI staining.Western blot was used to detect the protein expression of AKT and p-AKT.Results Compared to control group,three hours of hypoxia followed by sixteen hours of reoxygenation induced hippocampal neuronal apoptosis.EA could raise the neuronal viability and reduce apoptosis rate and the damage degree of rat hippocampal neurons.EA could increase the expressing of p-AKT.Conclusions EA has protective effects on damaged neurons,and the mechanism may be related to activating the PI3K-AKT signal transduction pathway.

Correlation of enhancement radiosensitization of emodin isolated from Guangxi P. multiflorum Thunb on hypoxic nasopharyngeal cancer cells with expression of DNA repair genes

Abstract: Aim To investigate the effects of emodin isolated from Guangxi P.multiflorum Thunb on the expression of KU70/KU80 in hypoxic nasopharyngeal cancer CNE-1 cells and reveal the relationship between radiosensitization of emodin monomer and DNA repair genes.Methods The expression of hypoxia inducible factor-1α(HIF-1α)and DNA double-strand break repair genes(KU70/KU80)between the experimental groups and the control group under hypoxic condition was detected by the real-time fluorescence quantitative RT-PCR.Results Expression of HIF-1α was significantly increased under hypoxia condition.HIF-1α had no change after treatment with emodin alone.The expression level of KU70/KU80 was induced by radiation.Compared with radiation alone group,radiation combined hypoxia group obviously enhanced the expression of KU70/KU80.KU70/KU80 mRNA expression significantly reduced after radiation combined with emodin under hypoxic condition.Conclusion In the hypoxic environment,emodin combined with radiotherapy can effectively inhibit the expression of HIF-1 α and DNA double-strand break repair genes(KU70/KU80),which may be its mechanism of radiosensitization.

Ampelopsin induces apoptosis via altering expression of Bcl-2/Bax and activating caspase-3 in human hepatoma cell line Bel-7402

Abstract: Aim To investigate the effects of ampelopsin on induction of apoptosis in human hepatocellular carcinoma Bel-7402 cells.Methods Bel-7402 cells were treated with ampelopsin with different concentrations for 24,48 and 72 h.The cell proliferation was detected by MTT assay.The morphological change of cells was observed through microscope observation by fluorescence staining.DNA fragmentation was visualized by agarose gel electrophoresis.The apoptosis rate was analyzed by flow cytometry.The expressions of caspase-3,Bcl-2 and Bax protein were detected by Western blot.Results Ampelopsin inhibited the proliferation of Bel-7402 cell line in a dose-and time-dependent manner.The IC50 values were 89.6±16.1,36.2±6.5 and 15.3±3.0 mg·L-1 at 24,48 and 72 h,respectively.The fluorescence microscope showed clearly cell apoptosis with apoptotic body.Agarose gel electrophoresis result showed that Bel-7402 treated with ampelopsin produced a DNA ladder band.The sub-G1 peak was detected and resulted in dose-and time-dependent increasing of the population of sub-G1 DNA content by flow cytometry.The expression of Bcl-2 protein was down-regulated,while the expression of Bax protein was up-regulated.The pro-caspase-3 protein was down-expressed and activated.Conclusions Ampelopsin could inhibit Bel-7402 proliferation through inducing cell apoptosis.The mechanism might be that ampelopsin could directly or indirectly enhance the level of anti-apoptosis protein Bcl-2 and decrease the level of apoptosis protein Bax.The pathway of pro-caspase-3 activated was initiated and effector caspase-3 was sequentially activated.

In vitro comparison of rotundine metabolism in liver microsomes of human,dog and rat

Abstract: Aim To investigate the inter-species differences of rotundine (RTD) metabolism in liver microsomes of human,dog and rat by comparing enzyme kinetics of the parent drug and the formation of its major metabolites.Methods The incubation systems of RTD with liver microsome of the three species were optimized in terms of RTD concentration and incubation time.The concentration of RTD in incubates was determined by a validated LC-MS method.The metabolites of RTD in incubates were separated and identified by LC-MS/MS.Results The biotransformation of RTD by human liver microsome was the slowest among the three species.The Km,Vmax,CLint and T12 of RTD obtained from human liver microsome were 2.67 μmol·L-1,0.095 μmol·L-1·min-1,14.6±0.91 ml·min-1·kg-1 and 298±18.0 min,respectively. The corresponding kinetic parameters for rat and dog liver microsomes were 3.24 and 5.31 μmol·L-1,0.122 and 0.228 μmol·L-1·min-1,87.5±2.79 and 139±1.43 ml·min-1·kg-1 71.0±2.30 and 62.3±0.647 min.Four isomeric metabolites were all found in the liver microsomes of three species,which were formed in incubates by O-demethylation at four methyl groups of RTD. It was observed that the relative amounts of these four metabolites generated in liver microsomes of three species were different.Conclusions The major metabolic pathway of RTD is O-demethylation in the liver microsomes from all three species.However,the inter-species differences are observed in terms of relative amounts of the metabolites measured in incubates,as well as in metabolic characteristics of RTD.

Mechanisms of curcumin protecting endothelial cells against ischemia and reperfusion injury

Abstract: Aim To investigate the mechanisms in protecting HUVEC against ischemia/reperfusion(I/R) injury directed by curcumin.Methods Hypoxia/reoxgenation(H/R) model was established on HUVEC.MTT colorimetric assay was used to observe the injury degree of hypoxia and reoxygenation at the different time.With preconditioning by different concentration of Cur,the survival rate of HUVEC subjected to H/R was assessed by MTT colorimetric assay.Pretreated with Cur(5 μmol·L-1),the expression of LC3,cathepsin B,cathepsin L,Bax and Bcl-2 were observed by fluorescent staining and Western blot in HUVEC during H/R process.Results Cur(1.25~5 μmol·L-1) played a protective role during H/R in HUVEC in a dose-dependent manner.During H/R,the expressions of LC3,cathepsin B and the ratio of Bax/Bcl-2 increased,and the nuclear translocation of cathepsin L was induced;when cur was pretreated,LC3 was furtherstrengthened,at the same time,the up-regulation of cathepsin B,the ratio of Bax/Bcl-2 and the nuclei-location of cathepsin L were inhibited partly by Cur.Conclusions Cur can raise the survival rate of HUVEC in the process of H/R.Cur increases the autophagy activity,depresses cathepsins and Bax/Bcl-2 to protect the endothelial cells.