中国中西医结合杂志2024，Vol.44Issue(2)：183-192,10.DOI:10.7661/j.cjim.20240110.098
从肝治心组方调控铁自噬保护缺血/再灌注损伤心肌细胞机制研究
Mechanism Study of Conggan Zhixin Decoction for Ischemia/Reperfusion Injury of Myocardial Cells Based on Ferritinophagy
摘要
Abstract
Objective To investigate the mechanism of improving myocardial ischemia/reperfusion injury using Conggan Zhixin Decoction(CGZXD).Methods Fifty SD rats were randomly divided into 5 groups:sham-operated group,model group,CGZXD group,Shexiang Baoxin Pill(SBP)group,Diltiazem group,with 10 rats in each group.A rat model of myocardial ischemia/reperfusion injury was prepared.The intervention group was given CGZXD[5.32 g/(kg·d)],SBP[10.27 mg/(kg·d)],and diltiazem[6.86 mg/(kg·d)]by gavage respectively.The sham-operated group and model group were given equal amounts of physiological saline by gavage.After 14 days of intervention,samples were taken,and the morphology of myocardial tissue was observed using HE staining.The morphology of myocardial cell mitochondria was observed using transmission electron microscopy.RT-PCR and Western Blot were used to detect the mRNA and protein expression of nuclear receptor co activator 4(NCOA4),ferritin heavy chain 1(FTH1),and prostaglandin G/H synthase 2(PTGS2),respectively.H9c2 rat cardiomyocytes were divided into 7 groups:normal group,model group,blank serum group,Chinese medicine group,ferroptosis inhibitor group(Ferrostatin-1),ferroptosis inducer group(Erastin),and Chinese medicine inducer group.After 12 hours of hypoxia and 2 hours of reoxygenation,the cell survival rate,reactive oxygen species(ROS)content,mitochondrial membrane potential level,and Fe2+level were measured in each group.The mRNA and protein expression of NCOA4,FTH1,and PTGS2 were detected by RT-PCR and Western Blot,respectively.Results Animal experiments showed that in the model group of rats,myocardial fibers were broken or sparse,with focal myocardial necrosis,interstitial infiltration of inflammatory cells,swelling and enlargement of mitochondria in myocardial cells,and proliferation of fibrous scar tissue.The pathological changes in the intervention group were alleviated.Compared with the sham-operated group,the mRNA and protein expression of NCOA4 and PTGS2 increased in the model group,while the mRNA and protein expression of FTH1 decreased(P＜0.05,P＜0.01).Compared with the model group,the mRNA and protein expression of NCOA4 and PTGS2 decreased in each intervention group,while the mRNA and protein expression of FTH1 increased(P＜0.05,P＜0.01).Cell experiments showed that compared with the normal group,the model group exhibited a decreasing in cell survival rate,membrane potential,FTH1 mRNAand protein expression,and an increasing in intracellular ROS and Fe2+levels,as well as NCOA4 and PTGS2 mRNA and protein expression(P＜0.05,P＜0.01).Compared with the model group,both the Chinese medicine group and the ferroptosis inhibitor group demonstrated an increasing in cell survival rate,membrane potential,FTH1 mRNA and protein expression,while intracellular ROS,Fe2+levels,NCOA4,and PTGS2 mRNAand protein expression decreased(P＜0.01).The cell survival rate,membrane potential,FTH1 mRNA and protein expression decreased in the ferroptosis inducer group,while intracellular ROS,Fe2+levels,NCOA4,PTGS2 mRNA,and protein expression increased(P＜0.05,P＜0.01).Compared with the ferroptosis inducer group,the Chinese medicine inducer group showed an increase in cell survival rate,membrane potential,FTH1 mRNA and protein expression,while the intracellular ROS,Fe2+levels,NCOA4,PTGS2 mRNA and protein expression decreased(P＜0.05,P＜0.01).Conclusion CGZXD may inhibit ferritinophagy by regulating the NCOA4/FTH1 signaling pathway,thereby reducing ferroptosis in cardiomyocytes and protecting ischemia/reperfusion-injured cardiomyocytes.关键词
从肝治心组方/缺血再灌注损伤/核受体共激活因子4/铁蛋白重链1/铁自噬/铁死亡/中药复方Key words
Conggan Zhixin Decoction/ischemia-reperfusion injury/nuclear receptor coactivator 4/ferritin heavy chain 1/ferritinophagy/ferroptosis/Chinese herbal compound引用本文复制引用
何飘,朴美虹,谢丽华,曾阳,王瑾茜,汪辛强,张程程,胡国恒,陈亚..从肝治心组方调控铁自噬保护缺血/再灌注损伤心肌细胞机制研究[J].中国中西医结合杂志,2024,44(2):183-192,10.基金项目
全国名老中医药专家传承工作室建设项目(No.国中医药函人教函[2022]75号) （No.国中医药函人教函[2022]75号）
湖南省自然科学基金资助项目(No.2021JJ40418) （No.2021JJ40418）
湖南中医药大学研究生创新课题立项项目(No.2022CX32) （No.2022CX32）
