摘要
目的探讨茯苓酸(Pac)是否通过调控PTEN诱导激酶1(PINK1)/Parkin RBR E3泛素蛋白连接酶(Parkin)信号通路介导线粒体自噬来改善冠心病(CHD)大鼠的心肌损伤。方法将SD大鼠分为对照(Con)组、CHD组、Pac低剂量组(Pac-L组)、Pac高剂量组(Pac-H组)、Pac高剂量+PINK1/Parkin信号通路抑制剂组(Pac-H+3-MA组),每组10只。除Con组外,其余各组大鼠建立CHD模型。造模成功后,各组大鼠腹腔注射相应药物或生理盐水。连续干预4周后,检测大鼠左室射血分数(LVEF)、左室舒张末容积(LVEDV)、左室收缩末容积(LVESV)、平均动脉压(MAP),血清中肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、肌钙蛋白I(cTnI)、肌钙蛋白T(cTnT)水平,心肌组织中肿瘤坏死因子α(TNF-α)、白细胞介素10(IL-10)、IL-1β、活性氧(ROS)、丙二醛(MDA)水平和过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性,以及p62、活化型胱天蛋白酶3(cleaved caspase-3)、Parkin、PINK1蛋白表达量与微管相关蛋白1轻链3Ⅱ型(LC3Ⅱ)/LC3Ⅰ比值;观察心肌组织形态和线粒体自噬囊泡,统计单位面积线粒体自噬囊泡数量和心肌细胞凋亡率。结果与CHD组比较,Pac-L组和Pac-H组大鼠的LVEF,MAP,IL-10水平,CAT、SOD活性,p62、Parkin、PINK1蛋白表达量,LC3Ⅱ/LC3Ⅰ值,单位面积线粒体自噬囊泡数量均显著升高(P<0.05);LVEDV、LVESV、CK-MB、LDH、cTnI、cTnT、TNF-α、IL-1β、ROS和MDA水平,细胞凋亡率,cleaved caspase-3蛋白表达量均显著降低(P<0.05);且Pac-H组各指标改变更显著(P<0.05);两组心肌组织病理形态均有不同程度改善。与Pac-H组比较,Pac-H+3-MA组大鼠上述指标均显著逆转(P<0.05)。结论Pac可能通过激活PINK1/Parkin信号通路促进CHD大鼠心肌细胞线粒体自噬,从而减轻炎症反应和氧化应激,改善心肌损伤。
OBJECTIVE To explore whether pachymic acid(Pac)regulates mitochondrial autophagy mediated by the PTENinduced kinase 1(PINK1)/Parkin RBR E3 ubiquitin-protein ligase(Parkin)signaling pathway to alleviate myocardial injury in coronary heart disease(CHD)rats.METHODS SD rats were divided into control(Con)group,CHD group,Pac low-dose group(Pac-L group),Pac high-dose group(Pac-H group),Pac-H+PINK1/Parkin signaling pathway inhibitor group(Pac-H+3-MA group),with 10 rats in each group.Except for the Con group,CHD models were established in the remaining groups of rats.After successful modeling,the rats in each group were intraperitoneally injected with the corresponding drugs or normal saline.After continuous intervention for 4 weeks,the left ventricular ejection fraction(LVEF),left ventricular end-diastolic volume(LVEDV),left ventricular end-systolic volume(LVESV),and mean arterial pressure(MAP)of the rats were detected.The levels of creatine kinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),cardiac troponin I(cTnI),and cardiac troponin T(cTnT)in the serum,as well as the levels of tumor necrosis factor-α(TNF-α),interleukin-10(IL-10),IL-1β,reactive oxygen species(ROS),malondialdehyde(MDA)in the myocardial tissue,and the activities of catalase(CAT)and superoxide dismutase(SOD),as well as the expression levels of p62,cleaved caspase-3,Parkin,PINK1 proteins and the ratio of microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ)/LC3Ⅰratio were measured.The morphology of myocardial tissue and mitochondrial autophagic vesicles were observed,and the number of mitochondrial autophagic vesicles per unit area and the rate of cardiomyocyte apoptosis were counted.RESULTS Compared with CHD group,LVEF,MAP,IL-10 levels,CAT and SOD activities,p62,Parkin,PINK1 protein expressions,LC3Ⅱ/LC3Ⅰratio,the numbers of mitochondrial autophagic vesicles per unit area in the Pac-L and Pac-H groups were increased significantly(P<0.05);the levels of LVEDV,LVESV,CK-MB,LDH,cTnI,cTnT,TNF-α,IL-1β,ROS and MDA,cell apoptosis rates,and protein expression of cleaved caspase-3 were all decreased significantly(P<0.05);and the changes in various indicators were more pronounced in the Pac-H group(P<0.05);both groups showed varying degree of improvement in myocardial histopathological morphology.Compared with the Pac-H group,the aforementioned indicators in rats from the Pac-H+3-MA group were all significantly reversed(P<0.05).CONCLUSIONS Pac may promote mitochondrial autophagy in cardiomyocytes of CHD rats by activating the PINK1/Parkin signaling pathway,thereby reducing inflammatory responses and oxidative stress and improving myocardial injury.
作者
谢建
高波
梁珊珊
杨清
郭思言
龚龙家
XIE Jian;GAO Bo;LIANG Shanshan;YANG Qing;GUO Siyan;GONG Longjia(Dept.of Cardiovas Medicine,Suizhou Hospital,Hubei University of Medical,Hubei Suizhou 441300,China)
出处
《中国药房》
北大核心
2025年第18期2267-2272,共6页
China Pharmacy
基金
湖北省自然科学基金项目(No.2023AFC049)。
