基于PINK1/Parkin通路探寻心复康口服液改善H9c2心肌细胞肥大的作用机制  

Exploring on Mechanism of Xinfukang Oral Liquid in Improving H9c2 Cardiomyocyte Hypertrophy Based on the PINK1/Parkin Pathway

作　　者：李家然 张笑涵 邱志凌 宋宇国 胡元会[1] 杜柏[1] Li Jiaran;Zhang Xiaohan;Qiu Zhiling;Song Yuguo;Hu Yuanhui;Du Bai(Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Bejing 100053,China)

机构地区：[1]中国中医科学院广安门医院,北京100053 [2]北京中医药大学,北京100029

出　　处：《中国中医急症》2025年第4期587-592,共6页Journal of Emergency in Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(82074409,82205097);中国中医科学院广安门医院引航工程临床类项目(HLCMHPP2023106)。

摘　　要：目的基于PTEN诱导假定激酶1(PINK1)/E3泛素连接酶(Parkin)通路探寻心复康口服液对H9c2心肌细胞肥大的改善作用及机制。方法通过20μmol/L苯肾上腺素(PE)干预H9c2细胞构建肥大模型,成功造模的细胞分别用最佳浓度的心复康含药血清(心复康组)和空白血清(模型组)干预,另设对照组。CCK-8法筛选最佳浓度;检测线粒体形态、转运孔蛋白20(Tom20)表达、线粒体膜电位、活性氧(ROS)含量、线粒体呼吸功能及三磷酸腺苷(ATP)水平;检测PINK1、Parkin、线粒体孔蛋白1(VDAC1)、1,4,5-三磷酸肌醇受体(IP3R)、葡萄糖调节蛋白75(Grp75)表达;检测B型利钠肽(BNP)、心房利钠肽(ANP)、PINK1、Parkin、胱天蛋白酶-3(Caspase-3)、胱天蛋白酶-9(Caspase-9)的mRNA表达。结果与对照组比较,模型组BNP、ANP表达、线粒体长度、ROS表达均升高(P<0.05);Tom20、线粒体膜电位、线粒体呼吸功能及ATP合成水平降低(P<0.01);PINK 1、Parkin蛋白及mRNA表达降低(P<0.01);VDAC1、IP3R、Grp75蛋白表达、Caspase-3/-9的mRNA表达升高(P<0.05)。与模型组比较,心复康组线粒体长度及ROS表达降低(P<0.05);Tom20、线粒体膜电位、呼吸功能及ATP水平上升(P<0.05);Parkin蛋白及mRNA表达升高(P<0.01);VDAC1、IP3R、Grp75蛋白表达、Caspase-3/-9的mRNA表达降低(P<0.05)。结论心复康口服液通过激活PINK 1/Parkin线粒体自噬通路,改善H9c2心肌肥大细胞线粒体的形态与功能,抑制心肌细胞凋亡。Objective:To explore the effect and mechanism of Xinfukang oral liquid on hypertrophy of H9c2 cardiomyocytes based on the PTEN-induced putative kinase 1(PINKI)/E3 ubiquitin ligase(Parkin)pathway.Methods:A hypertrophy model was established by treating H9c2 cells with 20μmol/L phenylephrine(PE).Successflly modeled cells were then treated with optimal concentrations of Xinfukang-contained serum(Xinfukang group)or blank serum(model group),with an additional normal control group.The optimal concentration was determined using the CCK-8 assay.Mitochondrial morphology,Tom20 expression,mitochondrial membrane potential,reactive oxygen species(ROS)content,mitochondrial respiratory function,and adenosine triphosphate(ATP)levels were measured.Protein expressions of PINK1,Parkin,voltage-dependent anion channel 1(VDAC1),inositol 1,4,5-trisphosphate receptor(IP3R),and glucose-regulated protein 75(Grp75)were detected.mRNA expressions of Btype natriuretic peptide(BNP),atrial natriuretic peptide(ANP),PINK1,Parkin,caspase-3,and caspase-9 were also measured.Results:Compared with the control group,the model group showed increased BNP and ANP expression,mitochondrial length,and ROS expression(P<0.05),decreased Tom20,mitochondrial membrane potential,mitochondrial respiratory function,and ATP synthesis levels(P<0.01),reduced PINK1 and Parkin protein and mRNA expression(P<0.01),and elevated VDAC1,IP3R,Grp75 protein expression,and caspase-3/-9 mRNA expression(P<0.05).Compared with the model group,the Xinfukang group exhibited decreased mitochondrial length and ROS expression(P<0.05),increased Tom20,mitochondrial membrane potential,respiratory function,and ATP levels(P<0.05),enhanced Parkin protein and mRNA expression(P<0.01),and reduced VDAC1,IP3R,Grp75 protein expression,and caspase-3/-9 mRNA expression(P<0.05).Conclusion:Xinfukang oral liquid ameliorates mitochondrial morphology and functions in hypertrophic H9c2 cardiomyocytes and inhibits cardiomyocyte apoptosis by activating the PINK1/Parkin mitophagy pathway.

关 键 词：H9C2心肌细胞 心复康口服液 PINK1/Parkin通路 线粒体功能 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]