参附益心颗粒对AngⅡ诱导的H9c2心肌细胞损伤及PGC-1α信号通路相关因子的影响  

Effects of Shenfu Yixin Granules (参附益心颗粒) on H9c2 Cardiomyocyte Injury and PGC-1α Signaling Pathway Related Factors Induced by AngⅡ

作　　者：刘倩茹 王新陆 朱明军 崔琳 李彬 王永霞 于瑞 LIU Qianru;WANG Xinlu;ZHU Mingjun;CUI Lin;LI Bin;WANG Yongxia;YU Rui(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou,450000;The First Affiliated Hospital of Henan University of Chinese Medicine)

机构地区：[1]河南中医药大学药学院,河南省郑州市450000 [2]河南中医药大学第一附属医院

出　　处：《中医杂志》2022年第7期679-687,共9页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82030120,81703897,82004178,81503419);河南省科技攻关计划(202102310177);河南省中医局课题(2019JDZX2015);河南省高校科技创新团队支持计划(13IRTSTHN012);河南省中原千人计划-中原名医项目。

摘　　要：目的 探讨参附益心颗粒治疗慢性心力衰竭可能的作用机制。方法 通过超高效液相色谱-四级杆-飞行时间串联质谱检测参附益心颗粒的离子成分。将H9c2心肌细胞分为正常1组、模型1组、参附益心颗粒低剂量组、参附益心颗粒高剂量组、氯沙坦组、辅酶Q10组。除正常1组外,其余各组采用血管紧张素Ⅱ(AngⅡ)诱导H9c2心肌细胞损伤模型,同时参附益心颗粒低、高剂量组分别用含参附益心颗粒0.25、0.5 mg/ml的培养基培养,氯沙坦组用含氯沙坦片1×10^(-4)mol/L的培养基培养,辅酶Q10组用含有辅酶Q10片1×10^(-4)mol/L的培养基培养,各组均培养24 h。再将H9c2细胞分为正常2组、模型2组、转染模型组、转染参附益心颗粒组、转染对照组。除正常2组、转染对照组外,其余各组采用AngⅡ诱导H9c2心肌细胞损伤模型,同时转染模型组、转染参附益心颗粒组、转染对照组给予Lipofectmine 2000转染试剂进行转染,转染参附益心颗粒组给予转染前实验结果更好的参附益心颗粒剂量继续培养。采用CCK-8法检测各组细胞活性,Mito-Tracker荧光探针观察H9c2细胞中线粒体含量,检测H9c2细胞中乳酸脱氢酶(LDH)含量及心肌细胞相关基因[过氧化物酶体增殖物激活受体α (PPARα)、过氧化物酶体增殖物激活受体β/δ (PPARβ/δ)、过氧化物酶体增殖物活化受体γ辅助激活因子1α (PGC-1α)、核呼吸因子1 (NRF-1)、核呼吸因子2 (NRF-2)、雌激素相关受体(ERR)] mRNA表达。结果 参附益心颗粒药物定性的正离子物质成分共有387个,含量最多的为Stachydrine (水苏碱)、5-Hydroxymethylfurfural (5-羟甲基糠醛)、Retronecine (倒千里光裂碱);定性的负离子物质成分共有308个,含量最多的为Danshensu(丹参素)、Malic acid(苹果酸)、4-hydroxybenzoic acid (4-羟基苯甲酸)。转染参附益心颗粒组给药剂量为0.5 mg/ml。基因沉默前,与正常1组比较,模型1组细胞活性及各相关�Objective To explore the mechanism of Shenfu Yixin Granules(参附益心颗粒)in the treatment of chronic heart failure(CHF).MethodsThe ionic components of Shenfu Yixin Granules were detected by ultra high performance liquid chromatography-quadruped-time-of-flight tandem mass spectrometry.H9c2 cardiomyocytes were divided into normal 1 group,model 1 group,Shenfu Yixin Granules low-dose group(SYL),Shenfu Yixin Granules high-dose group(SYH),losartan group,and coenzyme Q10 group.AngiotensinⅡ(AngⅡ)was used to induce H9c2cardiomyocytes injury model in all the groups except for the normal 1 group.Simultaneously,the cells in the SYL and SYH groups were cultured with the medium containing 0.25 mg/ml and 0.5 mg/ml of Shenfu Yixin Granules,respectively.The culture medium in the losartan group containd 1×10^(-4)mol/L of losartan tablets,and that in the coenzyme Q10 group used 1×10^(-4)mol/L of coenzyme Q10 tablets.Each group was cultured for 24 hours.The H9c2 cells were then divided into normal 2 group,model 2 group,transfection model group,transfection Shenfu Yixin Granules group(t-SY),and transfection control group.Except for the normal 2 group and the transfection control group,the other groups used AngⅡto induce H9c2 myocardial cell injury model.Simultaneously,the transfection model group,the t-SY group,and the transfection control group were given lipofectmine 2000 transfection reagent for transfection.The cells in the t-SYgroup was continuously cultured with Shenfu Yixin Granules using the dose that had better results before transfection.After that,the cell activity was detected by CCK-8.Mito-tracker fluorescent probe was used to detect mitochondrial content in H9c2 cells.The lactate dehydrogenase(LDH)content,and m RNA expression of cardiomyocyte-related genes including peroxisome proliferator-activated receptor alpha(PPARα),peroxisome proliferatoractivated receptorβ/δ(PPARβ/δ),peroxisome proliferator-activated receptor gamma coactivator 1α(PGC-1α),nuclear respiratory factor 1(NRF-1),nuclear respiratory

关 键 词：慢性心力衰竭 参附益心颗粒 基因沉默 PGC-1Α 乳酸脱氢酶 线粒体 

分 类 号：R285[医药卫生—中药学]