| Objective:This study used the principle and method of Cochrane systematic evaluation to evaluate the efficacy and safety of Jianpi Bushen Recipe in the treatment of Alzheimer's disease.In the pre-study of self-made "Bushen Yijing Recipe" by Prof.Yu in the treatment of Alzheimer's disease,there was positive effect in clinical efficacy and experimental research,but most of the experiments were on reinforcing kidney drugs.In the previous study in vitro,it proved that atractylodes decoction,one of the jianpi herbs of Jianpi Bushen Recipe,inhibited apoptosis.Therefore,this study investigated the effect of atractylenolide I,one of the main component of Atractylodes,on H2O2 mediated oxidative stress injury in bEnd.3 cells in vitro and its possible mechanism.Method:1.Evaluate the efficacy and safety of Jianpi Bushen Prescription in the treatment of Alzheimer's disease:using Meta-analysis,computer search Chinese database including China Biomedical Literature Database(CNM),China Journal Full-text Database(CNKI),Wip Database,Wanfang Database The search time was from 2010 to 2018.The validated data were extracted by searching,screening,and conforming to the documents that met the standards,and the results were systematically evaluated using Review Manager 5.2 software.Extract the valid data,through searching and screening,select the researches that meet the inclusive criteria,and use Review Manager 5.2 software to systematically evaluate the research results.2.To investigate the effect and mechanism of atractylenolide I on H2O2-mediated oxidative stress injury in bEnd.3 cells in vitro:The optimal experimental concentration of H2O2 and the suitable experimental concentration of atractylenolide I was screened by MTT method.To evaluate the effect of atractylenolide I,bEnd.3 cells were exposed to 400?M H2O2 for 6h after different concentrations of atractylenolide I pretreatment 24h.Cell viability was assessed using MTT method.To examine whether atractylenolide I could reverse the injury of H2O2-mediated on the bEnd.3 cells' viability by Hoechst33258 staining.The expressions of related apoptotic proteins Bax,Bcl-2,Cleaved caspase-3,Caspase-3,Cleaved caspase-9,Caspase-9,Cleaved caspase-12 and Caspase-12 were detected by Western Blot.Results:L.Eight randomized controlled trials were selected from 701 articles for systematic analysis.Meta-analysis results showed that the Jianpi Bushen Recipe group improved the Simple Mental State Scale(MMSE)and Daily Living Ability Assessment Scale(AD)in AD patients,and the difference was statistically significant.In terms of safety,3 of the 8 articles included in the study did not describe the adverse reactions,and the remaining 5 had no adverse reactions.2.The results of MTT assay showeed that:(1)bEnd.3 cells were treated with H2O2 concentration range of 100,200,400,600?mol/L for 6h,the viability inhibitory rate of bEnd.3 cells was close to 50%when H2O2 concentration was 400?mol/L.(2)Concentration range of atractylenolide I below 50 ?mol/L had no significant damage to bEnd.3 cells.(3)Results showed that H2O2 treatment significantly reduced the viability of bEnd.3 cells and that the high dose of atractylenolide I blocked the injury caused by H2O2.3.The results of Hoechst33258 staining showed that:(1)More dense and stained nucleus was observed after treatment with H2O2 400?mol/L,which showed high fluorescence.Atractylenolide I decreased the numbers of H2O2-induced high fluorescence cells.This suggest that atractylenolide I can inhibit H2O2-induced apoptosis.4.Western Blot results showed that:Ratio of Bax/Bcl-2,Cleaved caspase-12/caspase-12,Cleaved caspase-3/caspase-3,Cleaved caspase-9/caspase-9 decreased after pretreatment with high dose of atractylenolide I compared with the model group,Cyt-c expression decreased,the difference was statistically significant(P<0.05 or P<0.01).Conclusion:1.Based on the current clinical randomized controlled trials,it had been shown that Jianpi Bushen Recipe had a good curative effect on Alzheimer's disease.Because of the small amount of literature in this study,the safety of traditional Chinese medicine for treating Alzheimer's disease was unknown.2.By constructing a model of oxidative stress injury in vitro of H2O2 injured bEnd.3 cells,it was found that atractylenolide I inhibited H2O2-mediated apoptosis of bEnd.3 cells,and the regulation of expression of related apoptosis proteins as possible mechanism. |
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