参附注射液调控lncRNA ZFAS1介导线粒体自噬改善脓毒症心功能障碍的作用机制OA

Objective To investigate the mechanism of Shenfu Injection(SFI)in alleviating septic cardiac dysfunction by regulating long non-coding RNA(lncRNA)zinc finger antisense transcript 1(ZFAS1)-mediated mitophagy.Methods Seventy male C57BL/6J mice were randomly divided into a blank group,a model group,a Shenfu Injection(SFI)group(10 mL·kg-1),an SFI+ZFAS1 overexpression vector(pcDNA-ZFAS1)group,and an SFI+empty vector(pcDNA)group.The blank group contained 10 mice,and each of the remaining groups had 15 mice.After 7 consecutive days of administration,the mouse model of septic cardiac dysfunction was established by intraperitoneal injection of LPS.At 12 hours after modeling,cardiac function was evaluated by echocardiography[left ventricular ejection fraction(LVEF),left ventricular fractional shortening(LVFS),left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD)].Serum levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β were detected by ELISA.Pathological changes of myocardial tissue were observed by HE staining.Ultrastructural changes of myocardial mitochondria were observed by transmission electron microscopy.The mRNA expression levels of lncRNA ZFAS1,Beclin-1,microtubule-associated protein 1 light chain 3(LC3),PTEN-induced putative kinase 1(PINK1),and E3 ubiquitin-protein ligase Parkin in myocardial tissue were detected by RT-qPCR.The protein expression levels of PINK1,Parkin,LC3 Ⅱ/LC3 Ⅰ,and Beclin-1 were detected by Western blotting.Results(1)Compared with the blank group,the model group showed partial myocardial cell necrosis,structural disorder,cytoplasmic vacuolation,and extensive inflammatory infiltration;mitochondria were structurally disordered and swollen,with disappeared cristae and vacuole formation.Compared with the model group,the SFI group and SFI+pcDNA group showed reduced myocardial cell necrosis,more regular arrangement of muscle fibers,decreased inflammatory infiltration,and significantly reduced cytoplasmic vacuolation;mitochondria were relatively intact and orderly arranged,with clear cristae in most mitochondria and abundant autophagolysosomes.Compared with the SFI+pcDNA group,the SFI+pcDNA-ZFAS1 group showed obvious myocardial cell and mitochondrial damage,with reduced autophagy.(2)Compared with the blank group,the model group showed decreased LVEF and LVFS(P＜0.01),and increased LVEDD,LVESD,and serum levels of TNF-α,IL-6,and IL-1β(P＜0.05,P＜0.01).Compared with the model group,the SFI group and SFI+pcDNA group showed increased LVEF and LVFS(P＜0.01),and decreased LVEDD,LVESD,and serum levels of TNF-α,IL-6,and IL-1β(P＜0.05,P＜0.01).Compared with the SFI+pcDNA group,the SFI+pcDNA-ZFAS1 group showed decreased LVEF and LVFS(P＜0.01),and increased LVEDD,LVESD,and serum levels of TNF-α,IL-6,and IL-1β(P＜0.05,P＜0.01).(3)Compared with the blank group,the model group showed increased lncRNA ZFAS1 mRNA expression(P＜0.01)and significantly decreased PINK1,Parkin,LC3,and Beclin-1 mRNA and PINK1,Parkin,Beclin-1,LC3Ⅱ/LC3Ⅰ,and protein expression(P＜0.01)in myocardial tissue.Compared with the model group,the SFI group and SFI+pcDNA group showed decreased lncRNA ZFAS1 mRNA expression(P＜0.01)and significantly increased PINK1,Parkin,LC3,Beclin-1 mRNA and protein expression of PINK1,Parkin,LC3 Ⅱ/LC3 Ⅰ,and Beclin-1(P＜0.05,P＜0.01).Compared with the SFI+pcDNA group,the SFI+pcDNA-ZFAS1 group showed increased lncRNA ZFAS1 mRNA expression(P＜0.01),while the mRNA expression of PINK1,Parkin,LC3,Beclin-1,as well as the protein expression levels of PINK1,Parkin,Beclin-1 and LC3Ⅱ/LC3Ⅰ were significantly decreased(P＜0.05,P＜0.01).Conclusion Shenfu Injection may improve septic cardiac dysfunction in mice by downregulating lncRNA ZFAS1 expression and promoting cardiomyocyte mitophagy.