摘要
目的:探讨益母草碱(Leo)通过调节受体相互作用蛋白1(RIP1)-受体相互作用蛋白3(RIP3)-混合谱系激酶结构域样蛋白(MLKL)通路对心肌缺血/再灌注(MIR)损伤大鼠坏死性凋亡的影响。方法:随机取15只大鼠作为假手术组(Sham),其余大鼠结扎左冠状动脉前降支构建MIR模型后随机分为MIR组、Leo组(15 mg/kg)、Compound 6i组(1 mg/kg RIP1-RIP3-MLKL通路激活剂Compound 6i)、Leo+Compound 6i组(15 mg/kg Leo+1 mg/kg Compound 6i),每组15只。MIR组和Sham组注射等量生理盐水。超声心电图观察心功能指标;ELISA检测心肌损伤指标及炎症因子水平;HE染色观察心脏病理损伤情况;TTC染色检测心肌梗死面积;TUNEL染色检测细胞凋亡;Western blot检测RIP1-RIP3-MLKL通路蛋白表达。结果:Sham组大鼠心肌细胞完整,排列有序;MIR组心肌细胞排列紊乱和肿胀,肌原纤维收缩,肌膜被破坏,较Sham组射血分数(EF)及心输出量(CO)水平显著降低(P<0.05),Mb、cTnⅠ、CK-Mb含量、IL-6、IL-18水平、心肌梗死面积、心肌细胞凋亡率、RIP1、RIP3及MLKL蛋白水平显著升高(P<0.05);与MIR组相比较,Leo组细胞排列杂乱和水肿现象改善,EF、CO水平显著升高(P<0.05),Mb、cTnⅠ、CK-Mb含量、IL-6、IL-18水平、心肌梗死面积、心肌细胞凋亡率、RIP1、RIP3及MLKL蛋白水平显著降低(P<0.05),而Compound 6i组趋势相反;Leo+Compound 6i组与MIR组心肌组织结构相似,Compound 6i消除了Leo对MIR大鼠的心脏保护作用。结论:Leo可能通过下调RIP1-RIP3-MLKL通路缓解MIR大鼠心肌细胞坏死性凋亡,进而对MIR起治疗效果。
Objective:To investigate impact of leonurine(Leo)on necrotic apoptosis in rats with myocardial ischemia/reperfu-sion(MIR)injury by regulating receptor interacting protein 1(RIP1)-receptor interacting protein 3(RIP3)-mixed lineage kinase domain-like protein(MLKL)pathway.Methods:Fifteen rats were randomly selected as sham surgery group(Sham),left anterior descending coronary artery of remaining rats were ligated to construct MIR model,and randomly grouped into MIR group,Leo group(15 mg/kg),Compound 6i group(1 mg/kg RIP1-RIP3-MLKL pathway activator Compound 6i)and Leo+Compound 6i group(15 mg/kg Leo+1 mg/kg Compound 6i),with 15 rats in each group.MIR group and Sham group were injected with equal amounts of physiological saline.Cardiac function indicators were observed through ultrasound electrocardiogram;ELISA was applied to detect myocardial injury indicators and inflammatory factor levels;HE staining was applied to observe pathological damage of heart;TTC staining was applied to detect myocardial infarction area;TUNEL staining was applied to detect cell apoptosis;Western blot was applied to detect expres-sions of RIP1-RIP3-MLKL pathway proteins.Results:Myocardial cells in Sham group were intact and arranged in an orderly manner;arrangement of myocardial cells was disordered and swollen,myofibril contracted,and sarcolemma was destroyed in MIR group,ejec-tion fraction(EF)and cardiac output(CO)level were obviously lower than Sham group(P<0.05),Mb,cTnⅠ,CK-Mb contents,IL-6,IL-18 levels,myocardial infarction area,myocardial cell apoptosis rate,RIP1,RIP3 and MLKL protein levels were obviously increased(P<0.05);compared with MIR group,Leo group had improved cell arrangement disorder and edema,and obviously increased EF and CO levels(P<0.05),Mb,cTnⅠ,CK-Mb contents,IL-6,IL-18 levels,myocardial infarction area,myocardial cell apoptosis rate,RIP1,RIP3 and MLKL protein levels were obviously reduced(P<0.05),trend of Compound 6i group was opposite;myocardial tissue structure of Leo+Compound 6i group was similar to MIR group,and Compound 6i eliminated cardioprotective effect of Leo on MIR rats.Conclusion:Leo may alleviate myocardial necrosis apoptosis in MIR rats by down-regulating RIP1-RIP3-MLKL pathway,thus playing a therapeutic role in MIR.
作者
李明芳
唐碧波
杨才进
刘超权
LI Mingfang;TANG Bibo;YANG Caijin;LIU Chaoquan(Department of Pediatrics,Hainan Western Central Hospital,Danzhou 571700,China;Department of Cardiovascular Medicine,Hainan Western Central Hospital,Danzhou 571700,China)
出处
《中国免疫学杂志》
北大核心
2025年第8期1879-1884,共6页
Chinese Journal of Immunology
基金
2021年海南省科研课题(21A200309)。
