Vaspin通过AMPK/mTOR自噬信号通路影响2型糖尿病大鼠胰岛β细胞功能  

Effects of Vaspin on Pancreatic Beta Cell Function in Type 2 Diabetic Rats by AMPK/mTOR Autophagy Signaling Pathway

作　　者：魏姣姣 刘师伟 段瑞雪 李楠 王江娜 WEI Jiaojiao;LIU Shiwei;DUAN Ruixue;LI Nan;WANG Jiangna(The Ninth Clinical Medical College of Shanxi Medical University,Taiyuan 030009,China;Department of Endocrinology,Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Tongji Shanxi Hospital,the Third Hospital of Shanxi Medical University,Taiyuan 030032,China;Department of Endocrinology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;Department of Graduate School,Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学第九临床医学院,太原030009 [2]山西白求恩医院(山西医学科学院同济山西医院)山西医科大学第三医院内分泌科,太原030032 [3]华中科技大学同济医学院附属同济医院内分泌科,武汉430030 [4]山西医科大学研究生学院,太原030001

出　　处：《协和医学杂志》2023年第3期543-552,共10页Medical Journal of Peking Union Medical College Hospital

基　　金：山西省重点研发计划(201803D31133);山西省回国留学人员科研资助项目(2020-180);2020年山西省留学人员科技活动择优资助项目(20200032);中国糖尿病英才研究项目(2018-N-01-28)。

摘　　要：目的 探究内脏脂肪特异性丝氨酸蛋白酶抑制剂(visceral adipose tissue-derived serpin, Vaspin)改善2型糖尿病(type 2 diabetes mellitus, T2DM)大鼠胰岛β细胞功能的作用机制。方法 采用高脂高糖喂养联合腹腔注射链脲佐菌素的方式建立T2DM大鼠模型,并随机分为T2DM组(n=10)、Vaspin组(n=10),以同周龄正常饲料喂养的SD大鼠为正常对照组(n=10)。记录造模前及Vaspin干预前、干预4周和干预8周时3组大鼠体质量和空腹血糖(fasting blood-glucose, FBG)。Vaspin干预8周时,测定3组大鼠空腹胰岛素(fasting insulin, FINS)、糖耐量与胰岛素敏感性、胰岛β细胞功能及自噬相关蛋白表达水平,观察胰腺组织病理学形态。结果 与正常对照组比较,T2DM组与Vaspin组干预前、干预4周、干预8周时体质量均下降,FBG均升高(P均<0.05);与T2DM组比较,Vaspin组干预8周时大鼠体质量增高,FBG下降(P均<0.05)。组织病理示,正常对照组大鼠胰腺组织正常,胰岛细胞排列均匀、整齐,形态规则;T2DM组大鼠胰岛结构明显破坏,细胞分布不均匀、形状不规则;Vaspin组大鼠胰岛结构损伤、胰岛细胞形态破坏均较T2DM组减轻。干预8周时,与正常对照组比较,T2DM组及Vaspin组FINS降低,腹腔葡萄糖耐量试验(intraperitoneal glucose tolerance test, IPGTT)及腹腔胰岛素耐量试验(intraperitoneal insulin tolerance test, IPITT)血糖曲线下面积均升高(P均<0.05);与T2DM组比较,Vaspin组FINS升高,IPGTT与IPITT血糖曲线下面积均降低(P均<0.05)。高葡萄糖钳夹试验示,干预8周时,Vaspin组葡萄糖输注速率、第一时相及第二时相胰岛素分泌量虽低于正常对照组,但各指标均较T2DM组升高(P均<0.05)。免疫组化及Western blot结果示,干预8周时,与正常对照组比较,T2DM组大鼠胰腺组织中胰岛素表达水平及磷酸化哺乳动物雷帕霉素靶蛋白(phosphorylated mammalian target of rapamycin, p-mTOR)/mTOR比值均降低,P62、微管相关蛋白1轻链3(micObjective To investigate the mechanism of Vaspin in improving the pancreatic beta cell function in type 2 diabetic(T2DM)rats.Methods The diabetic rat model was established by feeding high⁃fat and high⁃sugar diet combined with intraperitoneal injection of streptozotocin.T2DM rats were randomly divided in⁃to T2DM group(n=10)and Vaspin group(n=10),with the same age SD rats fed with normal chow as normal control group(n=10).Before modeling,before Vaspin intervention,at 4 weeks and 8 weeks Vaspin intervention,the level of body weight and fasting blood⁃glucose(FBG)of rats were recorded.At 8 weeks of Vaspin intervention,fasting insulin(FINS),glucose tolerance and insulin sensitivity,pancreaticβ⁃cell function and autophagy⁃related protein expression levels were measured in three groups of rats,and histopathological mor⁃phology of the pancreas was observed.Results Before Vaspin intervention,at the 4 weeks and 8 weeks Vaspin intervention,compared with the normal control group,the level of body weight decreased and FBG increased in T2DM group and Vaspin group(all P<0.05).At 8 weeks Vaspin intervention,compared with T2DM group,the level of body weight increased and FBG decreased in Vaspin group(all P<0.05).Histopathology showed that the pancreatic tissue was normal,islet cells were arranged evenly,neatly and regularly in normal control group.In T2DM group,the islet structure was obviously destroyed,and the cells were unevenly distributed and irregularly shaped.Compared with T2DM group,the structural damage and morphological damage of islet cells in Vaspin group were significantly reduced.At the 8 weeks intervention,compared with the normal control group,the level of FINS decreased,while the area under the blood glucose curve of intraperitoneal glucose tolerance test(IPGTT)and intraperitoneal insulin tolerance test(IPITT)increased in T2DM group and Vaspin group(all P<0.05).Compared with T2DM group,the level of FINS increased,while the area under blood glucose curve of IPGTT and IPITT decreased in Vaspin group(a

关 键 词：内脏脂肪特异性丝氨酸蛋白酶抑制剂 2型糖尿病 腺苷酸活化蛋白激酶 哺乳动物雷帕霉素靶蛋白 自噬 胰岛Β细胞功能 

分 类 号：R587.1[医药卫生—内分泌] R365[医药卫生—内科学]