钠氢交换体1对糖尿病db／db小鼠心脏钙蛋白酶和糖原合成酶激酶-3β的调节作用  

Regulation of Na^＋-H ^＋ exchanger 1 on cardiac calpain and glycogen synthase kinase 3β in type 2 diabetic db/db mice

作　　者：李彩蓉 甘受益 黄凌萍 余辉 蒋学俊[1] 

机构地区：[1]武汉大学人民医院心内科,430060 [2]咸宁市中心医院心内科

出　　处：《中华糖尿病杂志》2017年第10期638-643,共6页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：国家自然科学基金（81470049）;湖北省教育厅重点项目（D20162803）

摘　　要：目的 探讨钠氢交换体1（NHE1）抑制剂卡立泊来德（CAR）对2型糖尿病db/db小鼠心脏钙蛋白酶（Calpain）和糖原合成酶激酶3β（GSK-3β）蛋白的调节作用.方法 选取7周龄雄性C57BL/KS db/db小鼠和野生型（WT）小鼠各16只,采用随机数字表法分为4组：对照组（WT）、CAR组（WT＋CAR）、模型组（db/db）和CAR干预组（db/db＋CAR）.10周后行血流动力学检查,测定空腹血糖（FPG）、甘油三酯（TG）和胆固醇（TC）及胰岛素,计算胰岛素抵抗指数（HOMA-IR）;观察心肌病理改变,测定心肌组织氧化/硝化应激反应;RT-PCR测定心肌组织磷酸烟酰胺腺嘌呤二核苷酸氧化酶p22phox和gp91phox亚基、转化生长因子β1（TGF-β1）、基质金属蛋白酶2（MMP-2）和基质金属蛋白酶9 （MMP-9）的mRNA水平.Western blotting检测心肌组织中钙蛋白酶1和2（Calpain-1和2）、钙调磷酸酶（CaN）、蛋白激酶B（AKT）和糖原合成酶激酶-3β（GSK-3β）蛋白的表达;并测定心肌Calpain和CaN酶的活性.两两比较采用t检验,组间比较采用单因素方差分析.结果（1）与db/db组比较,CAR＋db/db组小鼠FPG、TC、TG和HOMA-IR有下降趋势（t=1.213-2.112,P〉0.05）.（2）与WT组比较,db/db组心肌胶原含量、TGF-β1和MMP-2 mRNA水平明显增加,MMP-9 mRNA水平明显下降（t=3.695-4.905,P〈0.05）,而CAR干预后可逆转上述指标的改变（t=2.491-2.925,P〈0.05）.（3）CAR＋db/db组小鼠±dp/dtmax绝对值增大,LVEDP下降（t=2.865-6.404,P〈0.05）,并且小鼠心肌损伤情况明显改善.（4）与db/db组比较,CAR干预能明显改善db/db小鼠心脏组织中氧化/硝化应激,两组相比较有统计学差异（t=2.519-4.845,P〈0.05）.（5）与db/db组比较,CAR能降低db/db小鼠心肌Calpain、CaN酶活性和蛋白含量（t=2.634-3.253,P〈0.05）,还能上调p-AKT和p-GSK-3β蛋白表达（t=2.827,4.090,P〈0.05）.结论 CAR能改善db/db小鼠心脏损伤,其机制可能与CAR改善高糖高脂和胰Objective To explore the regulation of Na＋-H＋exchanger 1 inhibitor（Carporide,CAR） on cardiac Calpain and Glycogen synthase kinase 3β （GSK-3β） in type 2 diabetic db/db mice. Methods Sixteen 7 week old male C57BL/KS db/db mice and 16 wild-type mice were divided into four groups：Control group（WT）,CAR group（WT＋CAR）,Model group（db/db）and CAR treated group（db/db＋CAR）.The cardiac hemodynamics were measured after administration with CAR for 10 weeks. Fasting plasma glucose （FPG）, triglyceride （TG） and total cholesterol （TC） and insulin were determined and insulin resistance index （HOMA-IR） was calculated. The pathological changes of myocardium were observed and the oxidative/nitrifying stress responses were measured. The mRNA expressions of Nicotinamide-adenine dinucleotide phosphate（NADPH）oxidase p22phoxand gp91phox, transforming growth factor β1 （TGF-β1）, matrix metalloproteinase 2 （MMP-2） and matrix metalloproteinase 9 （MMP-9） were detected by Reverse Transcription-Polymerase Chain Reaction. The protein expressions of Calpain-1, Calpain-2, Calcineurin, AKT and GSK-3β were detected by western blotting. Results （1） Compared with db/db group, FPG, TC, TG and HOMA-IR decreased after CAR treatment in db/db mice（t=1.213-2.112,P〉0.05）.（2）Compared with the WT group, the collagen content and the mRNA levels of TGF-β1and MMP-2 in the db/db group increased significantly whereas the mRNA level of MMP-9 decreased significantly（t=3.098~4.905,P〈0.05）. Administration of CAR significantly reversed the changes（t=2.491-2.925,P〈0.05）.（3）Compared with WT mice,the LVEDP of mice in db/db group increased and the absolute value of ±dp/dtmaxdecreased which were significantly reversed by administration of CAR（t=2.865-6.404,P〈0.05）.（4）Compared with db/db group, the oxidative/nitrifying stress responses were significantly reversed by administration of CAR （t=2.519-4.845, P〈0.05）. （5） Compared with the db/db group, CAR can red

关 键 词：糖尿病心肌病 钠氢交换体1 卡立泊来德 钙蛋白酶 糖原合成酶激酶-3Β 

分 类 号：R741[医药卫生—神经病学与精神病学]