Objective:To investigate the protective effect of Danhong injection(DHI) on myocardial mitochondria in rats with coronary heart disease(CHD) and its regulation mechanism in homologous phosphatase tensin induced kinase 1(PINK1)/E3 ubiquitin ligase (Parkin) signaling pathway.Methods:The rat CHD model was constructed by continuous feeding of high-fat diet and free drinking water for 8 weeks.The intervention was performed with the low dose of Danhong injection(DHI-L) and high dose of Danhong injection(DHI-H).The function rescue experiment was performed with pcDNA3.1-PINK1(pc) which overexpressed PINK1.The proportion of intact mitochondria in myocardium of each group were detected by electron microscopy.T erminal-deoxynucleoitidyl transferase-mediated nick end labeling(TUNEL) was used to detect apoptosis in rat myocardial tissue.The content of reactive oxygen species(ROS) in rat myocardium was detected by superoxide anion dihydroethidine(DHE).The quantitative real-time polymerase chain reaction(qRT-PCR) was used to detect the mRNA expression of PINK1,Parkin,mitochondrial fusion protein 2(MNF2),mitochondrial division gene dynamic associated protein 1 ( DRP1),B-cell lymphoma-2 interaction protein 1 ( Beclin1),autophagy related gene 5 ( ATG5),and microtubule associated protein 1 light chain 3(LC-3) in myocardial tissue of rats in each group.The expressions of PINK1,Parkin,B-lymphoblastoma-2 (Bcl-2),and Bcl-2 associated X protein(Bax) in rat myocardial tissue were detected by Western Blot.Results:DHI-L and DHI-H could significantly increase the proportion of intact mitochondria in myocardial tissue of CHD rats,inhibit the apoptosis of cardiomyocyte,down-regulate the content of ROS in myocardial tissue,and decrease the mRNA expression of PINK1,Parkin,DRP1,Beclin1,ATG5,and LC-3 in myocardial tissue,increase the mRNA expression of MNF2 in myocardial tissue,down-regulate the expression of PINK1,Parkin and Bax in myocardial tissue,and up-regulate the expression of Bcl-2 in myocardial tissue,pc DNA3.1-PINK1 could partially reverse the protective effect of DHI on myocardial mitochondria,and the differences were statistically significant (P<0.05).Conclusion:Danhong injection can significantly inhibit oxidative stress in myocardial tissue of CHD rats,reduce the autophagy level of myocardial mitochondria,downregulate the apoptosis rate of cells in myocardial tissue,and improve the cardiac function of experimental animals,which may be related to the inhibition of PINK1/Parkin signal transmission.
coronary heart diseasemitochondrial damageDanhong injectionphosphatase and tensin homologue-induced putative kinase 1/E3 ubiquitin ligasesexperimental study
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