Ginsenoside Rg1 ameliorates OGD/R-induced PC12 cell injury by inhibiting autography via IRE1-JNK-CHOP pathway
This study investigated the protective effect of ginsenoside Rg1(GRg1)on oxygen and glucose deprivation/reoxygenation(OGD/R)-injured rat adrenal pheochromocytoma(PC12)cells and whether the underlying mechanism was related to the regulation of inositol-requiring enzyme 1(IRE1)-c-Jun N-terminal kinase(JNK)-C/EBP homologous protein(CHOP)signaling pathway.An OGD/R model was established in PC12 cells,and PC12 cells were randomly classified into control,model,OGD/R+GRg1(0.1,1,10 μmol·L-1),OGD/R+GRg1+rapamycin(autophagy agonist),OGD/R+GRg1+3-methyladenine(3-MA,autophagy inhibitor),OGD/R+GRg1+tunicamycin(endoplasmic reticulum stress agonist),OGD/R+GRg1+4-phenylbutyric acid(4-PBA,endoplasmic re-ticulum stress inhibitor),and OGD/R+GRg1+3,5-dibromosalicylaldehyde(DBSA,IRE1 inhibitor)groups.Except the control group,the other groups were subjected to OGD/R treatment,i.e.,oxygen and glucose deprivation for 6 h followed by reoxygenation for 6 h.Cell viability was detected by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide(MTT)assay.Apoptosis was detec-ted by Hoechst 33342 staining,and the fluorescence intensity of autophagosomes by the monodansylcadaverine(MDC)assay.Western blot was employed to determine the expression of autophagy-related proteins(Beclin1,LC3-Ⅱ,and p62)and the pathway-related pro-teins[IRE1,p-IRE1,JNK,p-JNK,glucose-regulated protein 78(GRP78),and CHOP].The results showed that GRg1 dose-de-pendently increased the viability of PC12 cells and down-regulated the expression of Beclin1,LC3-Ⅱ,p-IRE1,p-JNK,GRP78,and CHOP,compared with the model group.Furthermore,GRg1 decreased the apoptosis rate and MDC fluorescence intensity and up-regu-lated the expression of p62 protein.Compared with the OGD/R+GRg1(10 μmol·L-1)group,OGD/R+GRg1+rapamycin and OGD/R+GRg1+tunicamycin groups showed increased apoptosis rate and MDC fluorescence intensity,up-regulated protein levels of Beclin1,LC3-Ⅱ,p-IRE1,p-JNK,GRP78,and CHOP,decreased relative cell survival rate,and down-regulated protein level of p62.The 3-MA,4-PBA,and DBSA groups exerted the opposite effects.Taken together,GRg1 may ameliorate OGD/R-induced PC12 cell injury by inhibiting autophagy via the IRE1-JNK-CHOP pathway.
ginsenoside Rg1(GRg1)oxygen and glucose deprivation/reoxygenation(OGD/R)endoplasmic reticulum stressau-tophagyIRE1-JNK-CHOP
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