人分泌型Klotho蛋白对2型糖尿病心肌病大鼠的治疗作用及机制  

Therapeutic effect and mechanism of human secreted Klotho protein on type 2 diabetic cardiomyopathy rats

作　　者：张婷婷 左宪宏 王维娜 左晓霞 ZHANG Ting-Ting;ZUO Xian-Hong;WANG Wei-Na(School of Nursing of Zhangjiakou University,Zhangjiakou 075000,Hebei,China)

机构地区：[1]张家口学院护理学院,河北张家口075000 [2]张家口市第一医院心内科

出　　处：《中国老年学杂志》2024年第15期3684-3690,共7页Chinese Journal of Gerontology

基　　金：国家自然科学基金面上项目(81501764)。

摘　　要：目的探讨人分泌型Klotho蛋白对2型糖尿病心肌病大鼠的治疗作用及可能机制。方法40只大鼠给予高糖高脂饮食,链脲佐菌素(STZ)一次性腹腔注射诱导2型糖尿病心肌病模型,分为模型组、二甲双胍+苯那普利组(90+0.9 mg/kg)、低、高剂量组(50、100 mg/kg人分泌型Klotho蛋白)各10只;另取10只大鼠为正常组。各药物干预组于建模成功后给予相应药物灌胃,正常组及模型组给予等体积生理盐水,试验周期8 w。实验结束后,测定各组心功能指标及氧化损伤指标,实时荧光定量聚合酶链反应(RT-PCR)及Western印迹测定p-蛋白激酶B(AKT)、p-糖原合成酶激酶(GSK)-3βmRNA和蛋白水平。同时设心肌细胞(H9C2)组、棕榈酸(PAL)组、PAL+人分泌型Klotho蛋白组,测定各组细胞活力及p-AKT、p-GSK-3βmRNA和蛋白水平。结果与正常组比较,模型组、二甲双胍+苯那普利组与低、高剂量组左心室舒张末期内径(LVIDd)、左心室收缩末期内径(LVIDs)、丙二醛(MDA)明显升高,左心室短轴缩短率(LVFS)、左心室射血分数(LVEF)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)-Px、p-AKT、p-GSK-3βmRNA和蛋白表达明显降低(P<0.05)。与模型组相比,二甲双胍+苯那普利组与低、高剂量组LVIDd、LVIDs、MDA明显降低,LVFS、LVEF、SOD、GSH-Px、p-AKT、p-GSK-3βmRNA和蛋白表达明显升高,且人分泌型Klotho蛋白各剂量组效应具有明显剂量依赖性(P<0.05)。二甲双胍+苯那普利组和高剂量组各指标无明显差异(P>0.05)。与H9C2组比较,PAL组、PAL+人分泌型Klotho蛋白组光密度(OD)值、存活率、p-AKT、p-GSK-3βmRNA和蛋白表达明显降低(P<0.05);与PAL组比较,PAL+人分泌型Klotho蛋白组OD值、存活率、p-AKT、p-GSK-3βmRNA和蛋白表达明显升高(P<0.05)。结论人分泌型Klotho蛋白对2型糖尿病心肌病大鼠心功能损伤及心肌氧化损伤具有明显改善作用;其机制可能与人分泌型Klotho蛋白能促进p-AKT、p-GSK-3βmRNA�Objective To explore the therapeutic effect of human secreted Klotho protein on type 2 diabetic cardiomyopathy rats and its possible mechanism.Methods A total of 40 rats were given a high-sugar and high-fat diet,and the type 2 diabetic cardio-myopathy model was induced by one-time intraparitoneal injection of streptozotocin(STZ),and divided into model group,metformin+benazepril group(90+0.9 mg/kg),and low,high dose groups(50 and 100 mg/kg human secreted Klotho protein),with 10 rats in each group.Another 10 rats were selected as normal group.After successful modeling,each drug intervention group was given corre-sponding drug intragastric administration,normal group and model group were given equal volume normal saline.The test period was 8 w.After the experiment,cardiac function indexes and oxidative damage indexes of each group were determined.The mRNA and pro-tein levels of p-protein kinase B(AKT)and p-glycogen synthase kinase(GSK)-3βwere determined by real-time fluorescence quan-titative polymerase chain reaction(RT-PCR)and Western blotting.Cardiomyocytes(H9C2)group,palmitic acid(PAL)group,PAL+human secreted Klotho protein group were set up at the same time,and cell viability,p-AKT,p-GSK-3βmRNA and protein levels were measured in each group.Results Compared with normal group,left ventricular end diastolic diameter(LVIDd),left ventricular end diameter systolic(LVIDs)and malondialdehyde(MDA)in model group,metformin+benazepril group,low,high dose groups were significantly increased,left ventricular short axis shortening rate(LVFS),left ventricular ejection fraction(LVEF),superoxide dismutase(SOD),glutathione peroxidase(GSH)-Px,p-AKT,p-GSK-3βmRNA and protein expressions were significantly decreased(P<0.05).Compared with model group,LVIDd,LVIDs and MDA in metformin+benazepril group,low,high dose groups were signif-icantly decreased,LVFS,LVEF,SOD,GSH-Px,p-AKT,p-GSK-3βmRNA and protein expressions were significantly increased,the effects of each dose group of human secreted Klotho protein were significantly dose-de

关 键 词：人分泌型Klotho蛋白 糖尿病心肌病 蛋白激酶B(AKT)/糖原合成酶激酶(GSK)-3β通路 心功能 氧化损伤 

分 类 号：R587.2[医药卫生—内分泌]