N-乙酰氨基葡萄糖转移酶Ⅴ介导糖尿病小鼠心肌损伤的作用及机制  

Role and mechanism of GnT-Ⅴin modulation of cardiac damage in diabetic mice

作　　者：赵然 王婷婷 王晓宇 冯艳 李妍 ZHAO Ran;WANG Tingting;WANG Xiaoyu;FENG Yan;LI Yan(Department of Cardiology,Second Affiliated Hospital,Air Force Medical University,Xi′an 710068,China)

机构地区：[1]空军军医大学第二附属医院心血管内科,西安710068

出　　处：《山西医科大学学报》2024年第2期150-156,共7页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(82070385);唐都医院社会人才基金项目(2021SHRC017)。

摘　　要：目的 探究N-乙酰氨基葡萄糖转移酶Ⅴ(N-acetylglucosamine transferaseⅤ,GnT-Ⅴ)介导糖尿病小鼠心肌损伤的作用及其机制。方法 6~8周龄雄性C57BL/6小鼠30只,随机分为正常对照组(Ctrl组)、糖尿病组(DM组)和糖尿病+干扰GnT-Ⅴ基因的腺相关病毒组(DM+shGnT-Ⅴ组),每组10只。采用超声心动图检测各组小鼠心功能及左室质量,HE染色观察各组小鼠心肌组织病理变化,实时定量PCR检测心肌组织中心房钠尿肽(atrial natriuretic peptide, ANP)和B型钠尿肽(B-type natriuretic peptide, BNP)的表达,免疫印迹方法检测心肌组织中GnT-Ⅴ、转化生长因子-β激活激酶1(transforming growth factor-β-activated kinase 1, TAK1)蛋白表达。体外培养原代心肌细胞,分为正常葡萄糖组(NG组)、高糖组(HG组)、高糖+干扰GnT-Ⅴ基因的腺病毒组(HG+Ad-shGnT-Ⅴ组)和高糖+干扰GnT-Ⅴ基因的腺病毒+过表达TAK1基因的腺病毒组(HG+Ad-shGnT-Ⅴ+Ad-TAK1组)。免疫印迹方法检测原代心肌细胞中GnT-Ⅴ、TAK1蛋白表达,免疫荧光检测α-辅肌动蛋白(α-actinin)表达水平。结果 与Ctrl组相比,DM组小鼠心脏收缩功能显著下降(P<0.001),左室质量明显增加(P<0.001),心肌细胞肥大(P<0.001),GnT-Ⅴ和TAK1表达水平明显升高及ANP和BNP在mRNA水平表达显著增加(P<0.01)。与DM组相比,DM+shGnT-Ⅴ组小鼠心脏功能明显改善(P<0.01),左室质量明显降低(P<0.01),心肌肥大症状明显减轻(P<0.001),GnT-Ⅴ、TAK1、ANP及BNP表达明显减少(P<0.05)。与NG组相比,HG组原代心肌细胞α-actinin免疫荧光强度增强,GnT-Ⅴ和TAK1的表达显著增加(P<0.05);与HG组相比,HG+Ad-shGnT-Ⅴ组GnT-Ⅴ、TAK的表达显著减少(P<0.05),α-actinin免疫荧光强度减弱;与HG+Ad-shGnT-Ⅴ组相比,HG+Ad-shGnT-Ⅴ+Ad-TAK1组GnT-Ⅴ表达未见明显变化,TAK1的表达明显增加(P<0.05),心肌细胞α-actinin免疫荧光强度增强。结论 GnT-Ⅴ可能通过激活TAK1调控糖尿病小鼠的心肌损伤。Objective To investigate the role of N-acetylglucosamine transferaseⅤ(GnT-Ⅴ)in myocardial injury of diabetic mice and its underlying mechanism.Methods Thirty 6-8-week-old male C57BL/6 mice were randomly divided into control group(Ctrl group),diabetes mellitus group(DM group),and diabetes mellitus+AAV-shGnT-Ⅴgroup(DM+shGnT-Ⅴgroup),with 10 mice in each group.Echocardiography was used to detect the cardiac function and the left ventricular weight,HE staining was used to observe the pathological changes of myocardial tissues,and real-time quantitative PCR was used to detect the expressions of atrial natriuretic peptide(ANP)and B-type natriuretic peptide(BNP)in myocardial tissues.Western blot was used to detect GnT-Ⅴand TAK1 protein expressions in myocardial tissues.Primary cardiomyocytes were divided into normal glucose group(NG group),high glucose group(HG group),high glucose+Ad-shGnT-Ⅴgroup(HG+Ad-shGnT-Ⅴgroup)and high glucose+Ad-shGnT-Ⅴ+Ad-TAK1 group(HG+Ad-shGnT-Ⅴ+Ad-TAK1 group).Western blot was used to detect GnT-Ⅴand TAK1 protein expressions,and immunofluorescence was used to detect the level ofα-actinin expression.Results Compared with Ctrl group,the cardiac systolic function was decreased(P<0.001),the left ventricular mass was significantly increased(P<0.001),the diameters of cardiomyocytes were increased(P<0.001),and the expressions of GnT-Ⅴ,TAK1 and the mRNA levels of ANP and BNP were significantly elevated in DM group(P<0.05).Compared with DM group,the cardiac function was significantly improved in DM+shGnT-Ⅴgroup(P<0.01),the left ventricular mass was decreased(P<0.01),the symptoms of cardiac hypertrophy were significantly reduced(P<0.001),and the expressions of GnT-Ⅴ,TAK1,ANP and BNP were significantly reduced(P<0.05).Compared with NG group,α-actinin immunofluorescence intensity was significantly enhanced in HG group,and GnT-Ⅴ,TAK1 expressions were significantly increased(P<0.05).Compared with HG group,GnT-Ⅴand TAK1 expressions were significantly reduced in HG+Ad-shGnT-Ⅴgro

关 键 词：糖尿病 心肌肥厚 糖尿病心肌病 GnT-Ⅴ TAK1 

分 类 号：R587.1[医药卫生—内分泌]