炎症状态下西罗莫司对小鼠胰岛β细胞功能的影响  

Effect of sirolimus on pancreatic β cell function under inflammation condition

作　　者：王沛[1] 周维[1] 杨萍[1] 陈压西[1] 阮雄中[1] 赵蕾[1] 

机构地区：[1]重庆医科大学脂糖代谢性疾病重庆市重点实验室教育部感染性疾病分子生物学重点实验室,重庆400016

出　　处：《中国临床药理学杂志》2016年第10期909-912,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81200567;31540029;81400786)

摘　　要：目的研究炎症状态下西罗莫司对胰岛β细胞功能的影响。方法采用8周龄C57BL/6J雄性小鼠,随机分为A,B,C,D组:A组为对照组,小鼠隔天皮下注射生理盐水;B组小鼠隔天皮下注射酪蛋白;C组小鼠隔天皮下注射酪蛋白和溶媒介质;D组小鼠隔天皮下注射酪蛋白和西罗莫司。所有小鼠均给予高脂饮食喂养14周。通过葡萄糖耐量和胰岛素耐量了解小鼠体内糖代谢情况。酶联免疫吸附法(ELISA)检测小鼠血清中肿瘤坏死因子(TNF-α)、血清淀粉样蛋白(SAA)、胰岛素含量。用聚合酶链式反应(PCR)分析小鼠体内凋亡基因(BAX,BCL2)和胰岛素合成分泌相关基因的表达。苏木精-伊红(HE)染色观察小鼠胰岛形态,TUNEL染色观察胰岛细胞的凋亡情况。结果与A组相比,B组小鼠血清中炎症因子(TNF-α,SAA)显著增加(P<0.05),糖耐量及胰岛素耐量受损。葡萄糖耐量实验中,在30,60,120 min时D组小鼠血糖水平明显高于C组。胰岛素耐量实验中,D组小鼠血糖水平在15,30,60,120 min时明显高于C组。与C组相比,D组小鼠血清胰岛素含量明显下降(P<0.05),胰岛体积变小,胰岛β细胞凋亡增加,胰岛组织中BAX/BCL2增加,胰岛素合成分泌相关基因表达明显下降(P<0.05)。结论炎症状态下,西罗莫司使胰岛β细胞凋亡增加,胰岛素的产生和分泌下降,胰岛β细胞的功能降低,最终导致小鼠糖代谢紊乱。Objective To investigate the mechanism of sirolimus induced insulin resistance under inflammation stress.Methods Eight-week-old male C57 BL /6J mice were randomly assigned to A,B,C,D groups.Mice in group A received normal saline injection;group B received casein injection;group C received casein and vehicle injection;group D received casein and sirolimus injection.All injections were performed every other day.All mice were fed with a high-fat diet for 14 weeks.Then glucose tolerance test（ GTT） and insulin tolerance test（ ITT） were performed to test the glucose and insulin tolerances.The concentration of tumor necrosis factor（ TNF-α）,serum amyloid A（ SAA） and insulin in serum were measured by enzyme-linked immunosorbent assay（ ELISA）.Real-time PCR was used to determine the expression of apoptosis related gene（ BAX,BCL2） and key participants in β-cell functional integrity,such as pancreatic duodenal homeobox-1（ PDX1）,glucokinase（ GK）,glucose transporter 2（ GLUT2）.HE staining was used to observe the morphology of pancreatic islets.The apoptosis of pancreas was stained by TUNEL method.Results The contents of serum TNF-α and SAA in group B were much higher thanthose in group A.Compared with group C,group D showed higher glucose concentration in GTT（ 30,60,120 min）and ITT（ 15,30,60,120 min）.In group D,TUNEL staining showed more pancreatic β-cell apoptosis,BAX /BCL2increased（ P〈0.05）,the key genes participated in pancreatic insulin secretion were significantly reduced（ P〈0.05）.Compared with group C,the shape of islets was smaller to group D and serum insulin content much decreased（ P〈0.05）.Conclusions In inflammation condition,sirolimus treatment induced pancreatic β-cell apoptosis,reduced pancreatic β-cell function and decrease insulin secretion,thereby resulting in glucose metabolism disorders.

关 键 词：胰岛素抵抗 西罗莫司 炎症 胰岛Β细胞 凋亡 

分 类 号：R978.15[医药卫生—药品]