四逆汤抑制RhoA/ROCK信号通路改善大鼠心肌纤维化

摘要

目的 探究四逆汤(附子、干姜、甘草)对异丙肾上腺素诱导大鼠心肌纤维化的影响.方法 40只SD大鼠随机分为空白组、模型组、卡托普利组[100 mg/(kg·d)]、四逆汤组[3.8 g/(kg·d)],每组10只.除空白组外,其余大鼠皮下注射异丙肾上腺素[5 mg/(kg·d)]制备心肌纤维化模型,连续给药4周.末次给药后20 h,检测心脏血流动力学变化,计算心脏质量指数,苏木精-伊红(HE)染色及Masson染色观察心肌病理形态学变化,硝酸还原酶法检测血清中NO水平,黄嘌呤氧化酶法检测血清中超氧化物歧化酶(SOD)活力,硫代巴比妥酸法检测血清中丙二醛(MDA)水平,Western blot法检测心肌组织中RhoA、ROCK1、内皮型一氧化氮合酶(eNOS)蛋白的表达.结果 与模型组比较,四逆汤组大鼠心脏功能明显改善,心肌胶原含有量明显减少,血清中MDA水平明显降低,NO水平及SOD活力显著提高,心肌组织中RhoA和ROCK1蛋白的表达显著下降,eNOS蛋白的表达显著升高.结论 四逆汤能改善异丙肾上腺素诱导的大鼠心肌纤维化,其机制可能与抑制了RhoA/ROCK信号通路,进而上调了eNOS的表达后,导致氧化应激减少有关.%AIM To explore the effects of Sini Decoction (Aconiti lateralis Radix Praeparata,Zingiberis Rhizoma,Glycyrrhizae Radix et Rhizoma) on rat myocardial fibrosis induced by isoproterenol.METHODS Forty SD rats were randomly divided into blank,model,captopril [100 mg/(kg · d)] and Sini Decoction [3.8 g/(kg · d)] groups,with ten rats in each group.Except for the blank group,the rest rats were given subcutaneous injection of isoproterenol [5 rg/(kg · d)] to prepare myocardial fibrosis model,and successive administration lasted for four weeks.At 20 h after the last administration,hemodynamics changes of heart were detected;heart weight indexes were calculated;the changes of myocardial pathological morphology were observed by HE and Masson staining;NO level in serum was measured by nitrale reduetase;SOD activity in serum was measured by xanthinoxidase method;MDA level in serum was measured by thiobarbituric acid method;the protein expressions of RhoA,ROCK1 and eNOS in myocardial tissue were detected by Western blot method.RESULTS Compared with the model group,the cardiac function of rats was significantly improved;the myocardial collagen content was significantly decreased;the MDA level in serum was obviously decreased;the NO level and SOD activity were significantly increased;the protein expressions of RhoA and ROCK1 in myocardium tissue were significantly reduced,and the protein expression of eNOS was markedly increased in the Sini Decoction group.CONCLUSION Sini Decoction can improve the isoproterenol-induced myocardial fibrosis in rats,and its mechanism may be related to inhibiting the RhoA/ ROCK signaling pathway,in turn,raising the expression of eNOS,which leads to reduced oxidative stress.