芪参颗粒调控SIRT1/PGC-1α/FNDC5通路介导的线粒体稳态防治心力衰竭心室重构的作用机制  

Mechanism of Qishen Granules on preventing and treating ventricular remodeling in heart failure by regulating mitochondrial homeostasis mediated by SIRT1/PGC-1α/FNDC5 pathway

作　　者：王俊岩 董鑫 庄皓文 方春平[1] 谢抗 陈奕君 李春 王勇 王伟 WANG Jun-yan;DONG Xin;ZHUANG Hao-wen;FANG Chun-ping;XIE Kang;CHEN Yi-jun;LI Chun;WANG Yong;WANG Wei(College of Traditional Chinese Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405,China;School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]广州中医药大学中药学院,广州510006 [2]广州中医药大学第一附属医院,广州510405 [3]北京中医药大学中医学院,北京100029

出　　处：《中华中医药杂志》2023年第5期2292-2298,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82204990,No.82230126);中国博士后科学基金项目(No.2022T150149,No.2021M700966);广东省中医心脾相关病机和方药研究重点实验室(No.2022B1212010012)。

摘　　要：目的:探讨芪参颗粒调控SIRT1/PGC-1α/FNDC5通路介导的线粒体稳态防治心力衰竭心室重构的作用机制。方法:体内研究方面,通过胸主动脉结扎术构建慢性心力衰竭小鼠模型,并随机分为模型组,芪参颗粒低、中、高剂量组(简称低、中、高剂量组),培哚普利组,每组10只,另外10只假手术小鼠做假手术组。低、中、高剂量组造模4周后分别给予芪参颗粒1.42、2.83、5.66g·kg^(-1)·d^(-1)灌胃,培哚普利组给予培哚普利0.607 mg·kg^(-1)·d^(-1)灌胃,假手术组及模型组给予等量0.9%氯化钠溶液灌胃。连续给药8周后小动物超声观察各组小鼠心功能水平变化,苏木素伊红(HE)染色、Masson染色、WGA染色法观察各组小鼠心脏组织形态学变化及胶原纤维沉积情况。Western Blot法检测心脏组织SIRT1/PGC-1α/FNDC5通路及线粒体稳定相关蛋白表达。体外研究方面,培养HL-1心肌细胞,通过AngⅡ诱导心肌细胞损伤模型,并将细胞分为对照组、AngⅡ组、中药组,中药组给予10%芪参颗粒含药血清MEM培养基培养细胞,对照组及AngⅡ组给予10%普通大鼠血清MEM培养基培养,培养6 h后,各组细胞加入150 nmol/L AngⅡ孵育24 h。Western Blot法检测心肌细胞SIRT1/PGC-1α/FNDC5通路及线粒体稳定相关蛋白表达,免疫荧光法观察心肌细胞ROS及SIRT1、PGC-1α蛋白表达情况。结果:体内研究结果显示,与假手术组比较,模型组小鼠心功能显著降低(P<0.01),心肌细胞显著肥大,细胞间可见大量胶原纤维沉积,SIRT1、PGC-1α、FNDC5、Mitofusin 2、COXⅣ蛋白表达水平显著降低(P<0.01);与模型组比较,各治疗组小鼠心功能及相关蛋白表达水平显著改善(P<0.01,P<0.05)。体外研究结果显示,与对照组比较,AngⅡ组心肌细胞SIRT1、PGC-1α、FNDC5、Mitofusin 2、COXⅣ蛋白表达水平显著降低(P<0.01),ROS水平显著升高;经芪参颗粒含药血清预培养后各蛋白表达(P<0.01)及ROS水平均显著改善。结论:芪�Objective: To explore the mechanism of Qishen Granules on preventing and treating ventricular remodeling in heart failure by regulating mitochondrial homeostasis mediated by SIRT1/PGC-1α/FNDC5 pathway. Methods: In vivo, chronic heart failure mouse model was established by transverse aortic constriction(TAC) ligation and mice were randomly divided into model group, low, medium and high dose Qishen Granules groups(referred to as ‘low, medium and high dose group'), Perindopril group, with 10 mice in each group. Another 10 sham-operated mice were used as sham operation group. After 4 weeks, the low,medium and high dose groups were given Qishen Granules 1.42, 2.83 and 5.66 g·kg~(-1)·d^(-1) by gavage respectively. The perindopril group was given Perindopril 0.607 mg·kg~(-1)·d^(-1) by gavage. The sham operation group and the model group were given the same amount of normal saline by gavage. After 8 weeks of continuous administration, small animal ultrasound was used to observe the changes of cardiac function in each group of mice. Hematoxylin eosin(HE) staining, Masson staining and WGA staining were used to observe the morphological changes and collagen fiber deposition of heart tissue in each group. Western Blot was used to detect the expression of proteins related to the SIRT1/PGC-1α/FNDC5 pathway and mitochondrial stability in heart tissue. In vitro, HL-1 cardiomyocytes were cultured, and the cardiomyocyte injury model was induced by AngⅡ, and the cells were divided into control group, AngⅡ group and Chinese medicine group. Cells in the Chinese medicine group were cultured in MEM medium containing 10% Qishen Granules medicated serum. Cells in the control group and AngⅡ group were cultured in MEM medium containing 10% common rat serum. After 6 hours of culture, cells in each group were added with 150 nmol/L AngⅡ and incubated for 24 hours. Western Blot was used to detect the expression of proteins related to the SIRT1/PGC-1α/FNDC5 pathway and mitochondrial stability in cardiomyocytes. Results: In vivo

关 键 词：慢性心力衰竭 心室重构 线粒体稳态 SIRT1/PGC-1α/FNDC5 芪参颗粒 

分 类 号：R285.5[医药卫生—中药学]