养心康片调节自噬对慢性心力衰竭小鼠心肌纤维化的影响  

Effect of Yangxinkang Tablets in Regulating Autophagy on Myocardial Fibrosis in Mice After Chronic Heart Failure

作　　者：闫翠 周政[1] 梁碧荣 王陵军[1] 杨忠奇[1] 冼绍祥[1] YAN Cui;ZHOU Zheng;LIANG Bi-rong;WANG Ling-jun;YANG Zhong-qi;XIAN Shao-xiang(The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China)

机构地区：[1]广州中医药大学第一附属医院

出　　处：《中国实验方剂学杂志》2019年第3期53-58,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81673796,81473621);广东省自然科学基金重点项目(2016A030311030);广州市慢性心力衰竭中医药防治重点实验室项目(201705030006)

摘　　要：目的:研究养心康片对心衰后小鼠心肌纤维化的影响,探讨其作用机制。方法:采用胸主动脉缩窄法(TAC)建立小鼠慢性心衰模型,建模成功后随机分为假手术组,模型组,3-甲基腺嘌呤(3-MA)组(15 mg·kg^-1),养心康片高、中、低剂量组(1 170,585,390 mg·kg^-1),假手术组给予等体积蒸馏水灌胃。30 d后行心脏超声检查,收集血流动力学参数;通过心脏石蜡切片,马松(Masson)染色观察心肌细胞形态学变化及心肌纤维化程度;蛋白免疫印迹法(Western blot)检测微管相关蛋白轻链3(LC3),酵母ATG6同源物(Beclin-1),溶酶体膜蛋白(LAMP)心肌自噬蛋白,Ⅰ型胶原酶(CollagenⅠ),Ⅲ型胶原酶(CollagenⅢ),α-平滑肌肌动蛋白(α-SMA)心肌纤维化标志性蛋白表达情况。结果:与假手术组比较,模型组小鼠左心射血分数(LVEF),短轴缩短分数(FS)明显降低(P<0. 05),左心室舒张末期内径(LVDd),左心室收缩末期内径(LVDs)明显增大(P<0. 05),心肌纤维化程度显著加重(P<0. 01),α-SMA,CollagenⅠ,CollagenⅢ心肌纤维化标志性蛋白,LAMP,LC3,Beclin-1自噬蛋白表达均明显升高(P<0. 05);与模型组比较,3-MA组,养心康高、中剂量组心脏超声各项指标均显著改善(P<0. 05),心肌纤维化程度明显改善(P<0. 01),α-SMA,CollagenⅠ,CollagenⅢ,LAMP,LC3,Beclin-1表达明显降低(P<0. 05),低剂量组心功能、心肌纤维化及各项蛋白表达差异不明显。结论:养心康片可通过下调自噬减缓慢性心力衰竭小鼠心肌纤维化,改善心功能。Objective: To study the effect of Yangxinkang tablets on myocardial fibrosis in mice after heart failure,and to explore its mechanism. Method: The model of chronic heart failure in mice was established by thoracic aorta constriction( TAC). After successful modeling,mice were randomly divided into sham operation group,model group,3-methyladenine( 3-MA,15 mg·kg^-1) autophagy inhibitor group,Yangxinkang tablets high,medium,and low dose groups( 1 170,585,390 mg·kg^-1). The sham operation group received equal volume of distilled water. After 30 days,cardiac ultrasound was performed to collect hemodynamic parameters.Cardiac paraffin slices were stained with Masson to observe the morphological changes and fibrosis of cardiomyocytes. Western blot was used to detect lysosome-associated membrane protein( LAMP),microtubuleassociated protein light chain 3( LC3),Beclin-1 autophagyportein,α-smooth muscle activin( α-SMA),CollagenⅠ,Collagen Ⅲ protein expression. Result: As compared with normal group,the left ventricular ejection fraction( LVEF) and fractional shortening( FS) were significantly decreased( P< 0. 05),the left ventricular enddiastolic dimension( LVDd) and left ventricular end-systolic dimension( LVDs) were significantly increased( P<0. 05),the degree of myocardial fibrosis was significantly aggravated in model group( P< 0. 01),and the protein expression levels of α-SMA,CollagenⅠ,Collagen Ⅲ,LAMP,LC3,and Beclin-1 were significantly increased in model group( P< 0. 05). As compared with model group,the cardiac ultrasound indexes of the 3-MA group,Yangxinkang high and medium dose groups were significantly improved( P< 0. 05),the degree of myocardial fibrosis was significantly reduced( P< 0. 01),the protein expression levels of α-SMA,CollagenⅠ,CollagenⅢ,LAMP,LC3 and Beclin-1 were decreased in 3-MA group,Yangxinkang high and medium dose groups( P<0. 05),while the protein expression levels between the model group and Yangxinkang low-dose group showed no significant difference. Conclusion: Yangxinkang table

关 键 词：自噬 养心康片 慢性心力衰竭 心肌纤维化 Ⅰ型胶原酶(Collagen Ⅰ) Ⅲ型胶原酶(Collagen Ⅲ) α-平滑肌肌动蛋白(α-SMA) 

分 类 号：R289[医药卫生—方剂学] R541.61[医药卫生—中药学]