氧化苦参碱改善病毒性心肌炎小鼠 心室重构的作用研究

Effect of Oxymatrine on ventricular remodeling in mice with viral myocarditis

摘要:

目的 观察氧化苦参碱对病毒性心肌炎所致心力衰竭小鼠心室重构、心肌能量代谢的影响。 方法 将50 只雄性BALB/c 小鼠随机分为对照组、模型组（腹腔无菌注射0.1 ml 1×10-9 TCID50 柯萨奇B3 病毒），以及低、中、高剂量组（在模型组基础上分别皮下注射氧化苦参碱13、26 和52 mg/kg）。连续给药 14 d 后，采用高频超声心动图检测各组小鼠心功能；HE 染色、透射电镜观察左心室肌超微结构；TUNEL 法 检测各组小鼠左心室心肌细胞凋亡情况；ELISA 法检测左室心肌细胞游离脂肪酸（FFA）、三磷酸腺苷（ATP）、 腺苷酸（AMP）、乳酸（LAC）、Ⅰ型胶原蛋白（Col Ⅰ）、Ⅲ型胶原蛋白（Col Ⅲ）含量，Wsetern blotting 检 测各组小鼠左室心肌细胞葡萄糖调节蛋白78（GRP78）、C/EBP 环磷酸腺苷反应元件结合转录因子同源蛋白 （CHOP）、骨膜蛋白、钙联蛋白的表达。结果 模型组较对照组左室舒张末期内径扩大，左室射血分数和左 室短轴缩短率减小（P <0.05）；左室心肌细胞核排列紊乱、线粒体空泡化、肌丝断裂，出现大量病毒，心肌细 胞凋亡数量增多（P <0.05）；FFA、LAC、Col Ⅰ、Col Ⅲ、GRP78、CHOP、骨膜蛋白表达升高（P <0.05）； ATP/AMP 比值及钙联蛋白表达降低（P <0.05）。低、中、高剂量组较模型组左心室心肌细胞病理变化及左 室重构指标改善（P <0.05）；FFA、LAC、Col Ⅰ、Col Ⅲ、GRP78、CHOP、骨膜蛋白表达降低（P <0.05）； ATP/AMP 比值及钙联蛋白表达升高（P <0.05）。结论 氧化苦参碱可通过保护线粒体，改善心肌能量代谢， 减少心肌细胞凋亡，逆转心室重构，起到改善病毒性心肌炎小鼠心力衰竭的作用。

Abstract:

Objective To observe the effect of Oxymatrine on ventricular remodeling and myocardial energy metabolism in mice with congestive heart failure caused by viral myocarditis. Methods Fifty male BALB/c mice were randomly divided into a control group, a model group (intraperitoneal injection of 0.1 ml 1×10-9 TCID50 Coxsackie B3 virus), a low-dose group, a medium-dose group and a high-dose group (subcutaneous injection of Oxymatrine 13, 26 and 52 mg/kg into the model mice). After continuous administration for 14 d, cardiac function of each group was detected by high frequency echocardiography; structures and ultrastructures of left ventricular muscle were observed by HE staining and transmission electron microscopy; apoptosis of left ventricular cardiomyocytes was detected by TUNEL method; the levels of free fatty acids (FFA), adenosine triphosphate (ATP), adenosine monophosphate (AMP), lactic acid (LA), type I collagen (Col I) and type III collagen (Col III) in the left ventricular cardiomyocytes were detected by ELISA; Western blot was used to detect the expression of glucose-regulated protein 78 (GRP78) and C/EBP cyclic adenosine monophosphate response element binding transcription factor homologous protein (CHOP), periostin and calnexin in the left ventricular cardiomyocytes of mice. Results In the model group, LVEDD was enlarged, LVEF and FS were decreased (P < 0.05); the nuclei of the left ventricular cardiomyocytes were disordered, the mitochondria were vacuolated, the myofilaments were broken and the virus appeared, the number of myocardial cells was increased (P < 0.05); the expressions of FFA, LA, Col Ⅰ, Col III, GRP78, CHOP and periostin were increased (P < 0.05), the ratio of ATP/AMP and the expression of calnexin were decreased (P < 0.05). The pathological changes and left ventricular remodeling indexes of left ventricular cardiomyocytes in the lowdose, medium-dose and high-dose groups were improved (P < 0.05), the expressions of FFA, LA, Col Ⅰ, Col III, GRP78, CHOP and periostin were decreased (P < 0.05), the ATP/AMP ratio and calnexin expression were increased (P < 0.05). Conclusions Oxymatrine can ameliorate the heart failure in mice with viral myocarditis by protecting mitochondria, improving myocardial energy metabolism, reducing myocardial cell apoptosis and reversing ventricular remodeling.