摘要
目的:探讨双参宁心胶囊减轻大鼠心肌缺血再灌注损伤的机制是否与调控线粒体分裂、融合有关。方法:该研究以SD大鼠为研究对象,结扎冠状动脉左前降支,构建大鼠心肌缺血再灌注损伤模型,实验分为假手术组、模型组、双参宁心组(90 mg·kg^(-1))、曲美他嗪组(5.4 mg·kg^(-1))。微量法检测超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量,化学荧光法检测线粒体膜电位(△Ψm)变化、线粒体膜通透性开放孔(mPTP)开放程度,萤光素酶法检测细胞内腺嘌呤核苷三磷酸(ATP)水平,实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测线粒体分裂相关因子线粒体动力相关蛋白1(DRP1)、线粒体裂变蛋白1(FIS1),视神经萎缩症蛋白1(OPA1)、线粒体外膜融合蛋白1(MFN1)、MFN2的mRNA和蛋白表达水平。结果:与假手术组比较,模型组大鼠血清SOD活性降低,MDA含量升高,△Ψm、mPTP开放水平、ATP含量均降低,线粒体分裂因子DRP1、FIS1蛋白表达升高,融合相关因子OPA1、MFN1的mRNA转录和蛋白表达均降低;与模型组比较,双参宁心胶囊可显著升高大鼠血清SOD活性,降低MDA含量,升高细胞内ATP水平及△Ψm,降低mPTP开放水平,下调线粒体分裂因子DRP1、FIS1蛋白表达,升高融合相关因子OPA1、MFN1的mRNA转录和蛋白表达水平。结论:双参宁心胶囊减轻大鼠心肌缺血再灌注损伤的机制与调控线粒体分裂、融合有关,其具体机制可能为抑制线粒体分裂因子DRP1、FIS1表达从而抑制线粒体分裂,升高融合相关因子OPA1、MFN1表达,进而保护线粒体功能结构减轻心肌缺血再灌注损伤。
Objective:To explore whether the mechanism of Shuangshen Ningxin capsules(SSNX) in alleviating myocardial ischemia-reperfusion injury(MIRI) in rats is related to the regulation of mitochondrial fission and fusion.Method:This study focused on Sprague Dawley(SD) rats and ligated the left anterior descending branch of the coronary artery to construct a rat model of MIRI.The rats were divided into the sham operation group,model group,SSNX group(90 mg·kg^(-1)) and trimetazidine group(5.4 mg·kg^(-1)).The activity of superoxide dismutase(SOD) and the content of malondialdehyde(MDA) were detected by micro method.Changes in mitochondrial membrane potential ( △ Ψm) and the degree of mitochondrial permeability transition pore(mPTP) opening were detected by the chemical fluorescence method.The intracellular adenosine triphosphate(ATP) level was detected by the luciferase assay.The messenger ribonucleic acid(mRNA) and protein expression levels of mitochondrial fission and fusion related factors dynamin-related protein 1(DRP1),mitochondrial fission 1 protein(FIS1),optic atrophy protein 1(OPA1),mitochondrial outer membrane fusion protein 1(MFN1),and MFN2 were detected by real-time polymerase chain reaction(real-time PCR) and Western blot.Result:Compared with the sham operation group,the model group showed a decrease in serum SOD activity and an increase in MDA content.The opening level of mPTP,the level of △Ψm and ATP content decreased,the protein expressions of mitochondrial fission factors DRP1 and FIS1 increased,and the protein expressions and mRNA transcription levels of fusion related factors OPA1 and MFN1 decreased.Compared with the model group,SSNX significantly increased serum SOD activity,reduced MDA content,increased intracellular ATP level and △Ψm,reduced the opening level of mPTP,downregulated the protein expressions of mitochondrial fission factors DRP1 and FIS1,and increased the mRNA transcription levels and protein expressions of fusion related factors OPA1 and MFN1.Conclusion:SSNX inhibits the express
作者
辛高杰
陈原原
刘子馨
尤越
曹策
王奥奥
孟红旭
韩笑
刘建勋
李磊
付建华
XIN Gaojie;CHEN Yuanyuan;LIU Zixin;YOU Yue;CAO Ce;WANG Aoao;MENG Hongxu;HAN Xiao;LIU Jianxun;LI Lei;FU Jianhua(Institute of Basic Medicine,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;National Clinical Research Center for Chinese Medicine Cardiology,Beijing 100091,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2024年第7期87-94,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82174219,82174015)
中国中医科学院科技创新工程项目(CI2021A00912)。
关键词
双参宁心胶囊
线粒体分裂
线粒体融合
心肌缺血再灌注损伤
Shuangshen Ningxin capsules
mitochondrial fission
mitochondrial fusion
myocardial ischemia-reperfusioninjury
