The Epigenetic Regulation Of BDNF Was Involved In The Protective Effect Of Three Active Ingredients Of Cistanche On PC12 Cells Damage Induced By A?_25-35

Objective: To investigate the protective effect of OC1?OC4 and CDPS of Fomes officinalis Cistanche on the ?-amyloid protein(A?25-35)induced Rat adrenalpheochromocytoma(PC12)injury;to investigate the effects of three active components of Cistanche deserticola on PC12 cells injury induced by A?25-35,and to study the effect of histone acetylation on bdnf gene transcription.Methods:(1)The cell survival rate determined by MTT method and determine the A?25-35 induced PC12 nerve cell damage model of dose and action time.The cultured PC12 cells were divided into ten groups: normal control group,model group(A?25-35 injury group),OC1(50?g/mL?150 ?g/mL),OC4(50?g/mL?100?g/mL),CDPS(50?g/mL?150?g/m L),TSA(5nmol/L),GSrd(20 ?g/mL).(2)We performed Hoechst 333258 staining to observe the nuclear morphological changes associated with apoptosis afer giving OC1 ?OC4?CDPS?TSA and GSrd;V-FITC and PI double staining flow cytometry apoptosis.(3)ELISA method to detect HATs?HDAC2 and BDNF levels.(4)Western Blotting detection P300 and HDAC2 protein expression level.(5)The alterations of histone H3 acetylation and HDAC2 in BDNF exon ? were observed and analyzed by chromatin immunoprecipitation(ChIP).Results:(1)After exposure of the PC12 cells 48 h by 20umol/LA?25-35,determined by MTT test cell survival rate(42.91±4.59)%,meaning has a significant difference compared with the control group(P<0.05),exposure to 20?M A?25–35 alone significantly increased early(LR)and late(UR)apoptotic cell death in PC12 cells.However,preincubation with OC1?OC4?CDPS resulted in a significant decrease in the apoptotic rate.shows that successful cell aging model was established.(2)Compared with model group,each group HATs expression increase s,HDAC2 decrease,and BDNF levels increase,and there are dose dependent.(3)Compared with the model group,HDAC2 blocker TSA group and each administration group after48 h,ChIP analysis confirmed that A?25-35 induced BDNF exon ? histone H3 acetylation protein levels decreased significantly,the reduction of histone acetylation is HDAC2-dependent.Conclusion: Acteoside?crenotaside and CDPS Cretone glycoside,had significant protective effects on PC12 cells exposed to A?25-35.The mechanism may be that the decrease of acetylation of HDAC2-dependent histone H3 induced the suppression of continuous transcription of bdnf gene.Cretone glycoside,crenotaside and cistanche deserticola could activate BDNF gene transcription by inhibiting the expression of HDAC2 to protect PC12 cells and stimulate the regeneration of PC12 cells.