芪参还五胶囊通过抑制心肌坏死性凋亡改善小鼠急性心肌损伤的实验研究

An Experimental Study on the Improvement of Acute Myocardial Injury in Mice by Qishen Huanwu Capsules Through Inhibiting Necroptosis of Cardiomyocytes

目的观察芪参还五胶囊改善吡柔比星致小鼠急性心肌损伤并探寻其作用机制。方法12周龄雄性C57BL/6小鼠30只,随机分为对照组、模型组及芪参低、中、高剂量组。通过吡柔比星造模,取材前称量体重并通过心脏超声检测左心室结构和功能,ELISA检测血清心肌肌钙蛋白(cTnI)和氨基端前脑钠素(NT-proBNP)含量;HE染色评价小鼠心肌组织形态学改变;RT-qPCR检测心肌组织中焦亡相关基因NOD样受体热蛋白结构域相关蛋白3(NLRP3)、消皮素D(GSDMD)、半胱氨酸天冬氨酸蛋白酶1(Caspase1)和坏死性凋亡相关基因白细胞介素-1α(IL-1α)、白细胞介素-33(IL-33)、肿瘤坏死因子-α(TNF-α)、高迁移率族蛋白B1(HMGB1)的mRNA水平;Weston blotting检测焦亡相关蛋白NLRP3、GSDMD、Cleaved-Caspase1和坏死性凋亡相关蛋白磷酸化混合谱系激酶样蛋白(MLKL)、磷酸化受体相互作用蛋白激酶3(RIP3)、HMGB1水平。结果与对照组相比,模型组小鼠精神状态萎靡,体重下降(P<0.05),射血分数(LVEF)下降(P<0.05),血清c TnI和NT-proBNP水平升高(P<0.05),心肌损伤评分升高(P<0.05),坏死性凋亡相关基因和蛋白水平均有升高(P<0.05);与模型组相比,芪参还五胶囊可以改善小鼠精神状态,增加体重(P<0.05),升高LVEF(P<0.05),降低血清中cTnI和NT-proBNP水平,改善心肌损伤(P<0.05),降低坏死性凋亡相关基因及蛋白表达(P<0.05);5组间焦亡相关基因及蛋白表达无明显差异(P>0.05)。结论芪参还五胶囊可以改善吡柔比星造成的小鼠急性心肌损伤,其机制可能与抑制心肌细胞坏死性凋亡有关。

Objective:To investigate the effect of Qishen Huanwu Capsules on acute myocardial injury caused by pirarubicin in mice and its underlying mechanism.Methods:30 male 12-week-old C57BL/6 mice were randomly divided into control group,model group,low-dose group,medium-dose group,and high-dose group.Acute myocardial injury was induced by pirarubicin.Before tissue collection,body weight was measured,and left ventricular structure and function were assessed using echocardiography.Serum levels of cardiac troponin I(cTnI)and Nterminal pro-brain natriuretic peptide(NT-proBNP)were detected by ELISA.Histological changes in the myocardium were evaluated by HE staining.RT-qPCR was used to detect mRNA levels of pyroptosis-related genes[NOD-like receptor protein 3(NLRP3),gasdermin D(GSDMD),caspase-1(Caspase1)]and necroptosis-related genes[interleukin-1α(IL-1α),IL-33,tumor necrosis factor-α(TNF-α),high mobility group box 1(HMGB1)]in myocardial tissue.Western blotting analysis was performed to detect protein levels of pyroptosis-related proteins(NLRP3,GSDMD,Cleaved-Caspase-1)and necroptosis-related proteins[phosphorylated mixed lineage kinase domain-like protein(MLKL),phosphorylated receptor-interacting serine/threonine-protein kinase 3(RIP3),HMGB1].Results:Compared with the control group,the model group showed decreased activity,reduced body weight(P<0.05),decreased ejection fraction(LVEF)(P<0.05),increased serum cTnI and NT-proBNP levels(P<0.05),and elevated myocardial injury scores(P<0.05);levels of necroptosis-related genes and proteins were also increased(P<0.05).Compared with the model group,Qishen Huanwu Capsules improved the general condition of the mice,increased body weight(P<0.05),increased LVEF(P<0.05),reduced serum cTnI and NT-proBNP levels,and ameliorated myocardial injury(P<0.05).The expression of necroptosis-related genes and proteins was also decreased(P<0.05).There were no significant differences in the expression of pyroptosis-related genes and proteins among the 5 groups(P>0.05).Conclusion:Qishen Huanwu Capsules can improve acute myocardial injury caused by pirarubicin,and this effect may be related to the inhibition of necroptosis in cardiomyocytes.