Study effects and mechanism of PHB on reactive oxygen species and apoptosis in cardiomyocytes under high glucose / 中国免疫学杂志

ABSTRACT

Objective:To investigate the effect and mechanism of PHB on the apoptosis of H 9C2 cardiomyocytes under high glucose.Methods:Select pCDNA3-PHB and pCDNA3-NC recombinant plasmid transfect into cardiomyocytes and effected by high glu-cose.The experiment were divided into four groupscontrol group,high glucose group(HG group),high glucose+empty vector group (HG+pCDNA3-NC group),high glucose+pCDNA3-PHB plasmid group(HG+pCDNA3-PHB group).The expression of PHB,Bax/Bcl-2,c-caspase3,AKt and p-AKt protein after transfected were detected by Western blot .ROS were detected under high glucose by DHE staining.The TNF-αmRNA and IL-6 mRNA were detected by Real-time quantitative PCR(qRT-PCR).Flow cytometry was used to detect the apoptotic rate .Results:The recombinant plasmid pCDNA 3-PHB could increase the expression of PHB .The apoptotic rate of HG group,HG+pCDNA3-NC group and HG+pCDNA3-PHB group were significantly higher than control group (P<0.05).Compared with HG group,the apoptotic rate of HG+pCDNA3-NC group was not significant(P>0.05),but that of HG+pCDNA3-PHB group was significantly lower(P<0.05).The levels of ROS,TNF-αmRNA,IL-6 mRNA,Bax/Bcl-2 and c-caspase3(P<0.05) protein in the HG group,HG+pCDNA3-NC group,HG+pCDNA3-PHB group were significantly higher than the control group .Compared with HG group , the level of ROS,TNF-αmRNA,IL-6 mRNA,Bax/Bcl-2 and c-caspase3 (P>0.05)protein in the HG+pCDNA3-NC group were no significant difference,but the ROS,TNF-αmRNA,IL-6 mRNA、Bax/Bcl-2 and c-caspase3 (P<0.05)protein in the HG+pCDNA3-PHB group were significantly decreased .Statistical analysis showed that there was no significant difference of AKt protein in each group (P>0.05).The relative expression of p-AKt protein in HG group,HG+pCDNA3-NC group and HG+pCDNA3-PHB group were signif-icantly lower than that in control group ( P<0.05 ) ,HG+pCDNA3-NC group was no significantly different than HG group ( P>0.05 ) , However ,HG+pCDNA3-PHB group was significantly higher than HG group ( P<0.05 ) .Conclusion: PHB overexpression can inhibit the apoptosis of H9C2 cardiomyocytes in high glucose environment ,andreduce the inflammatory response by effected the regulation of PI3K/Akt signaling pathway,reduced ROS,Bax/Bcl-2,c-caspase3 protein levels,that give us a new ideas and methods of DCM .