Effect of Shenfu Injection of Anti-myocardial Ischemia-Reperfusion Injury via Modulating HIF-1 α/BNIP3 Pathway-Mediated Mitophagy
Objective To explore the mechanism of Shenfu Injection in regulating mitophagy to alleviate myocardial ischemia-reperfusion injury(MIRI)in rats.Methods Fifty rats were randomly divided into 5 groups,i.e.,the sham group,the model group,the low,medium,and high dose Shenfu Injection group,10 rats in each.The rats were administered before modelling.The rats in the Shenfu Injection groups were administered intraperitoneally(3.34,6.68,and 13.36 mL·kg-1·d-1),the rats in the sham group and the model group were injected intraperitoneally with saline once a day for 7 consecutive days to establish model.Twenty-four hours after the last administration,the anterior descending branch of the left coronary artery was ligated for 30 min,and then reperfused for 120 min to establish the MIRI model.The cardiac structure and cardiac function were assessed by ultrasound.The levels of serum creatine kinase(CK-MB),lactate dehydrogenase(LDH),and troponin(cTnl)were detected by ELISA.The infract size was determined by TTC.The changes of pathological in myocardial tissues were assessed by HE staining.Western Blot and qRT-PCR were used to detect the protein and mRNA expression of hypoxia-inducible factor 1α(HIF-1α),Bcl-2 interacting protein 3(BNIP3),microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ),and microtubule-associated protein 1 light chain 3-Ⅰ(LC3-Ⅰ).The apoptosis rate of cardiomyocytes were detected by TUNEL method.The number of mitochondrion and autophagosome were observed by electron microscopy.Results Compared with the sham group,the left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),and left ventricular end-diastolic diameter(LVEDd)decreased(P<0.05),left ventricular end-systolic diameter(LVESd)increased(P<0.05),infract size increased(P<0.01),the levels of CK-MB,LDH,and cTnl increased(P<0.05),necrosis and inflammatory infiltration in cardiomyocytes increased;the expressions of HIF-1α,BNIP3,and LC3-Ⅱ/LC3-Ⅰ ratio increased(P<0.05),HIF-1α,BNIP3,LC3 mRNA expressions increased(P<0.05);cardiomyocyte apoptosis rate,the number of clearly damaged mitochondria and autophagosomes per unit area increased(P<0.01,P<0.05)in the model group.Compared with the model group,the LVEF,LVFS,LVEDd,and LVESd in the Shenfu Injection groups increased(P<0.01).The expressions of HIF-1α,BNIP3,and LC3 Ⅱ/LC3 Ⅰ protein and mRNA increased in Shenfu Injection groups,except LC3Ⅱ/LC3 Ⅰ ratio reduced in the high dose Shenfu Injection group(P<0.01,P<0.05);infract size reduced(P<0.01),and the CK-MB,LDH,cTnl content decreased(P<0.05),cardiomyocyte apoptosis rate,mitochondrial damage and the number of autophagosomes reduced Shenfu Injection groups(P<0.01,P<0.05).Conclusion Shenfu Injection could protect the myocardium of rats with MIRI,and its mechanism might be related to promoting of HIF-1α/BNIP3 pathway-mediated mitophagy.
Shenfu Injectionmyocardial ischemia-reperfusion injuryhypoxiainducible factor 1αBcl-2 interacting protein 3mitophagyChinese patent medicine