Antagonistic Effects Of Paeoniflorin On H 2 O 2 -induced Endoplasmic Reticulum Stress In H9C2 Cardiomyocytes

Objective:1.To investigate the effect of paeoniflorin on H9C2 myocardial cells;2.To prepare H9C2 myocardial cells injury model induced by H₂O₂ and investigate the effect of paeoniflorin on H9C2 cells injury model induced by H₂O_2;3.To invest igate the role of endoplasmic reticulum stress in paeoniflorin antagonizing oxidative stress injury.Methods:1.Cultured H9C2 myocardial cells in vitro,and treat myocardial cells with a concentration of 12.5μM,25μM,50μM,100μM,200μM,400μM paeoniflorin for12h,24h,and the survival rate of cells was detected by MTT;treat myocardial cell swith a concentration of 0μM,100μM,200μM,400μM,800μM,1600μM H₂O₂ for 4h,a-nd the survival rate of cells was detected by MTT;2.H9C2 myocardial cells were preconditioned by paeoniflorin with a concentration of 50μM,100μM,200μM for 20h,then adding the final concentration of 400μM H₂O₂ for 4h,and the survival rate of cells was detected by MTT,and lactate dehydrogenase（LDH）leakage,malondial dehyde（MDA）content and superoxide dismutase（SOD）activity were assayed by ch emical reagent kit;3.H9C2 myocardial cells were pretreated with a concentration of5mM endoplasmic reticulum stress inhibitor,4-Phenyl butyric acid（4-PBA）,200μM paeoniflorin and 4-PBA combined with paeoniflorin for 24h,adding a final concent ration of 400μM H₂O₂ for 4h,and the survival rate of cells was detected by MTT methodand the expression of BiP/GRP78,CHOP were detected by Western blot me thod.Results:1.Compared with normal control group,the survival rate of small conce-ntration range（12.5μ²00μM）of paeoniflorin groups were higher,and there is a conc entration dependent manner（P<0.05）,and the survival rate of high concentration（400μM）paeoniflorin group was obviously lower（P<0.05）;the survival rate of every H2O2group were decreased,and there is a concentration dependent manner（P<0.05）;2.Co mpared with the H₂O₂ model group,the survial rate of paeoniflorin intervention gro up were higher,and SOD activity of paeoniflorin intervention group were significa ntly increased in a dose-dependent manner（P<0.05）;compared with normal control group,SOD activity of H2O2 model group were significantly decreased（P<0.05）.Co mpared with the H2O2 model group,LDH leakage of paeoniflorin intervention grou p were significantly decreased in a dose-dependent manner（P<0.05）;compared with normal control group,LDH leakage of the H₂O₂ model group were significantly in creased（P<0.05）;compared with the H₂O₂ model group,MDA contenet of paeoniflor in intervention group were significantly decreased in a dose-dependent manner（P<0.05）;compared with normal control group,MDA content of the H2O2 model group were sig-nificantly increased（P<0.05）.3.Compared with normal control group,the ex pression o-f BiP/GRP78 and CHOP of H₂O₂ model group were increased（P<0.05）,a nd the survial rate were decreased;compared with the H₂O₂ model group,the expr ession of Bi P/GRP78 and CHOP of paeoniflorin intervention group and 4-PBA inte rvention group were decreased（P<0.05）;the difference in expression of BiP/GRP78and CHOP of paeoniflorin inte-rvention group and 4-PBA intervention group were not obvious（P>0.05）,and the difference in survival rate was not obvious;the expres sion of Bi P/GRP78 and CHOP of paeoniflorin combined with 4-PBA group were l ess than paeo-niflorin intervention group and 4-PBA intervention group（P<0.05）,the cell survival rate was increased.Conclusions:1.Paeoniflorin can increase the survival rate of H9C2 cells and pr o-mote the proliferation of H9C2 cells in a certain concentration range（12.5μ²00μM）;2.H2O2 can reduce the activity of myocardial cells and inhibit cell proliferation and paeoniflorin can protect against oxidative stress injury induced by H₂O₂ in H9C2 cells;3.Paeoniflorin may inhibit endoplasmic reticulum stress,which play an imp ortant role in antagonizing oxidative stress injury.