竹节参总皂苷调控AMPK/mTOR/ULK1通路介导的铁自噬对糖尿病心肌病大鼠心肌细胞铁死亡的抑制作用  

Total saponins of Panax japonicus regulates the inhibitory effect of AMPK/mTOR/ULK1 pathway mediated ferritinophagy on ferroptosis of cardiomyocytes in diabetic cardiomyopathy rats

作　　者：任晓静 张海龙 程远 马媛 Ren Xiaojing;Zhang Hailong;Cheng Yuan;Ma Yuan(Department of Endocrinology,Zhumadian Central Hospital,Zhumadian 463000,China;Clinical Pharmacy Room,Zhumadian Central Hospital,Zhumadian 463000,China)

机构地区：[1]驻马店市中心医院内分泌科,463000 [2]驻马店市中心医院临床药学室,463000

出　　处：《中华内分泌代谢杂志》2024年第1期53-63,共11页Chinese Journal of Endocrinology and Metabolism

摘　　要：目的探究竹节参总皂苷(TSPJ)对糖尿病心肌病(DCM)大鼠心肌细胞铁死亡的作用及调控机制。方法实验1:将SD大鼠分为对照组、DCM组、TSPJ低剂量组、TSPJ高剂量组、二甲双胍组(Met),每组各10只。实验2:将SD大鼠分为对照组、DCM组、TSPJ组、腺苷酸活化蛋白激酶(AMPK)抑制剂Compound C组、TSPJ+AMPK激动剂AICAR组,每组各10只。除对照组外,其余各组大鼠均采取腹腔注射链脲佐菌素构建DCM模型。各组大鼠给予对应药物干预8周后,检测各组大鼠体重及糖脂代谢水平;超声心动图检测大鼠心功能指标;酶联免疫吸附(ELISA)法检测各组大鼠血清乳酸脱氢酶(LDH)、心肌肌钙蛋白I(cTnI)、肌酸激酶同工酶MB(CK-MB)水平;苏木精-伊红(HE)染色和电镜观察心肌组织病理变化;试剂盒检测心肌组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)、丙二醛、活性氧簇(ROS)及Fe2+水平;Western印迹法检测心肌组织铁死亡、铁自噬以及AMPK/哺乳动物雷帕霉素靶蛋白/UNC-51样激酶1(mTOR/ULK1)信号通路相关蛋白表达。结果与对照组相比,DCM组大鼠左心室射血分数(EF)、左室短轴缩短率(FS)、舒张早期与晚期血流速度峰值比值(E/A)均显著降低,舒张末期左心室内径(LVEDd)增加,血清LDH、cTnI、CK-MB升高,心肌组织SOD、GSH降低,丙二醛、ROS和Fe2+增加,转铁蛋白受体1(TFR1)、核受体共激活因子4(NCOA4)、LC3-II/LC3-I、Beclin-1、磷酸化AMPK及磷酸化ULK1表达水平升高,谷胱甘肽过氧化物酶4(GPX4)、溶质载体家族7成员11(SLC7A11)、磷酸化mTOR表达水平降低。与DCM组相比,各治疗组大鼠上述指标均得到显著改善。与TSPJ组相比,AMPK激动剂AICAR能够逆转TSPJ对DCM大鼠心肌细胞铁死亡及AMPK/mTOR/ULK1通路介导的铁自噬作用。结论 TSPJ能够通过调控AMPK/mTOR/ULK1介导的铁自噬抑制DCM大鼠心肌细胞铁死亡,改善心肌损伤。Objective To investigate the effect of total saponins of Panax japonicus(TSPJ)on ferroptosis of myocardial cells in diabetic cardiomyopathy(DCM)rats and underlying mechanism.Methods Experiment 1:SD rats were divided into control group,DCM group,low-dose TSPJ group,high-dose TSPJ group,and metformin(Met)group,with 10 rats in each group.Experiment 2:SD rats were divided into control group,DCM group,TSPJ group,adenosine monophosphate-activated protein kinase(AMPK)inhibitor Compound C group,and TSPJ+AMPK agonist AICAR group,with 10 rats in each group.Except for the control group,all rats were intraperitoneally injected with streptozotocin to construct a DCM model.After 8 weeks of corresponding drug intervention,the body weight as well as glucose and lipid metabolism of rats in each experimental group were assessed,and the cardiac function indicators were detected with echocardiography.The levels of serum lactate dehydrogenase(LDH),cardiac troponin I(cTnI)and creatine kinase isoenzyme MB(CK-MB)were detected by ELISA technique.The pathological changes of myocardial tissue were observed using hematoxylin-eosin(HE)staining.The levels of dismutase(SOD),glutathione(GSH),malondialdehyde(MDA),reactive oxygen species(ROS)and Fe2+in myocardial tissue were detected.Western blot was used to detect ferroptosis,ferritinophagy,and the AMPK/mammalian target of rapamycin/UNC-51-like kinase 1(mTOR/ULK1)signaling pathway related proteins expression in myocardial tissue.Results Compared with control group,left ventricular ejection fraction(EF),left ventricular short axis shortening rate(FS),peak blood velocity ratio(E/A)between early and late diastolic periods were significantly decreased in DCM group,left ventricular inner diameter(LVEDd)was increased,and the serum LDH,cTnI,CK-MB were increased,the levels of SOD,GSH were decreased,MDA,ROS,Fe2+were increased in myocardial tissue,the expressions of TFR1,NCOA4 LC3-II/LC3-I,Beclin-1,phosphorylated AMPK and phosphorylated ULK1 were increased,the expressions of GPX4,SLC7A11 and phosphorylate

关 键 词：糖尿病心肌病 竹节参总皂苷 铁死亡 铁自噬 AMPK/mTOR/ULK1通路 

分 类 号：R285.5[医药卫生—中药学]