心阳片通过抑制AGEs介导的心脏成纤维细胞激活改善心力衰竭小鼠心肌纤维化的作用机制

Study on the Mechanism of Xinyang Tablets in Improving Myocardial Fibrosis in Mice with Heart Failure by Inhibiting AGEs-Mediated Activation of Cardiac Fibroblasts

目的探讨心阳片(吉林参、淫羊藿、黄芪、益母草、毛冬青等)通过抑制晚期糖基化终产物(Advanced glycation end-products,AGEs)介导的心脏成纤维细胞激活改善心力衰竭小鼠心肌纤维化的作用机制。方法将48只C57BL/6J小鼠随机分为假手术组、模型组、培哚普利组(0.61 mg·kg^(-1))及心阳片低、中、高剂量组(455、910、1820 mg·kg^(-1)),每组8只。采用横向主动脉缩窄(Transverse aortic constriction,TAC)法建立心力衰竭小鼠模型。造模成功后,按照上述剂量灌胃给药,每天1次,连续6周。采用超声心动图检测小鼠心功能:左室射血分数(LVEF)、左室短轴缩短率(LVFS);免疫组织化学法测定小鼠心脏组织AGEs的表达;qPCR法检测小鼠心脏组织中vimentin、FSP1及纤维化相关基因collagen Ⅰ、collagen Ⅲ、fibronectin、CTGF的mRNA表达;Western Blot法检测小鼠心脏组织中vimentin蛋白的表达;Masson染色法观察小鼠心肌纤维化程度。结果与假手术组比较,模型组小鼠的LVEF、LVFS显著降低(P<0.01);小鼠心脏组织中的AGEs表达显著上调(P<0.01),vimentin、FSP1 mRNA表达及vimentin蛋白表达明显上调(P<0.05,P<0.01);心脏组织中有大量胶原纤维沉积,collagen Ⅰ、collagen Ⅲ、fibronectin、CTGF mRNA表达均显著上调(P<0.01)。与模型组比较,心阳片低、中、高剂量组及培哚普利组小鼠的LVEF、LVFS均明显升高(P<0.05,P<0.01);小鼠心脏组织中的AGEs表达均显著下调(P<0.01),vimentin、FSP1 mRNA表达及vimentin蛋白表达均明显下调(P<0.05,P<0.01);心脏组织中无明显胶原纤维沉积,collagenⅠ、collagen Ⅲ、fibronectin、CTGF mRNA表达均明显下调(P<0.05,P<0.01)。结论心阳片可以有效改善心力衰竭小鼠的心肌纤维化,可能与其抑制AGEs介导的心脏成纤维细胞激活有关。

Objective To investigate the mechanism of Xinyang Tablets(composed of Jilin Ginseng Radix et Rhizoma,Epimedii Folium,Astragali Radix,Leonuri Herba,Ilex Pubescens Radix,etc.)in improving myocardial fibrosis in mice with heart failure by inhibiting advanced glycation end-products(AGEs)-mediated activation of cardiac fibroblasts.Methods Forty-eight C57BL/6J mice were randomly divided into sham-operation group,model group,Perindopril group(0.61 mg·kg^(-1))and Xinyang Tablets low-,medium-and high-dose groups(455,910 and 1820 mg·kg^(-1)),with 8 mice in each group.The transverse aortic constriction(TAC)method was used to establish the heart failure mice model.After successful modeling,the drug was administered by gavage at the above dose once a day for 6 consecutive weeks.Echocardiography was used to detect the cardiac function of mice,including left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS);immunohistochemistry was used to determine the expression of AGEs in mice heart tissue;qPCR was used to detect mRNA expresstions of vimentin,FSP1 and fibrosis-related genes collagen Ⅰ,collagen Ⅲ,fibronectin and CTGF in mice heart tissue;the expression of vimentin protein in heart tissue was detected by Western Blot;the degree of myocardial fibrosis in mice was detected by Masson staining.Results Compared with the sham-operation group,LVEF and LVFS were significantly decreased in the model group(P<0.01);the expression of AGEs was significantly up-regulated in the heart tissue of mice(P<0.01),and the mRNA expressions of vimentin,FSP1and protein expression of vimentin were significantly up-regulated(P<0.05,P<0.01);there was a large amount of collagen fiber deposition in the heart tissue,and mRNA expressions of collagen Ⅰ,collagen Ⅲ,fibronectin and CTGF were significantly up-regulated(P<0.01).Compared with the model group,the LVEF and LVFS of mice in the low-,medium-and high-dose groups of Xinyang Tablets and Perindopril group were significantly increased(P<0.05,P<0.01);the expression of AGEs in the heart tissue of mice was significantly down-regulated(P<0.01),and the mRNA expressions of vimentin,FSP1 and protein expression of vimentin were significantly downregulated(P<0.05,P<0.01);there was no significant collagen fiber deposition in heart tissue,and mRNA expressions of collagen Ⅰ,collagen Ⅲ,fibronectin and CTGF were significantly down-regulated(P<0.05,P<0.01).Conclusion Xinyang Tablets can effectively improve myocardial fibrosis in mice with heart failure,which may be related to its inhibition of AGEs-mediated activation of cardiac fibroblasts.