梁露群,任飞燕,刘玲伶,彭伟,张小欢,毛彦稳,刘慧铭,王圆圆,石明隽,郭兵
LIU Luqun,REN Feiyan,LIU Lingling,PENG Wei,ZHANG Xiaohuan,MAO Yanwen,LIU Huiming,WANG Yuanyuan,SHI Mingjun,GUO Bing

• 1:贵州医科大学基础医学院病理生理学教研室
• 2:贵州医科大学贵州省常见慢性疾病发病机制及药物研究重点实验室

摘要(Abstract)：

目的:观察控制血糖前后Ski蛋白在糖尿病(DM)大鼠肾组织的表达变化,探讨其在糖尿病肾病(DN)发病过程中的作用。方法:24只SD大鼠随机分为正常组(NC)、DM组和胰岛素治疗(INS)组,DM组和INS组采用链脲佐菌素复制DM大鼠模型,INS组于造模成功4周后给予胰岛素22 U/(kg·d)治疗6周,于造模后第10周,采用生化法测定糖化血红蛋白(HbA1c)、血糖及24 h尿蛋白量,观察肾组织病理变化,免疫组织化学染色检测肾组织Ski、Collagen-Ⅲ及E-cadherin表达,Western Blot检测SD大鼠肾组织中Ski、转化生长因子β1(TGF-β1)、Collagen-Ⅲ、Smad3及p-Smad3(Ser432/425)蛋白的表达水平。结果:与NC组大鼠相比,DM组大鼠体质量减轻、血糖及Hb A1c增高,肾小管和肾小球纤维化改变明显,INS组上述改变有所改善;免疫组织化学染色显示,肾组织Ski、Collagen-Ⅲ表达从高到低依次为DM组>INS组>NC组(P<0.05),E-cadherin表达从高到低依次为NC组>INS组>DM组(P<0.05); Western Blot检测结果显示,肾组织TGF-β1、Collagen-Ⅲ、p-Smad3(Ser 432/425)和Ski蛋白的表达从高到低依次为DM组>INS组>NC组(P<0.05)。结论:DM大鼠肾组织Ski蛋白表达增高、TGF-β1/Smad3通路活化导致肾脏纤维化;而控制血糖后,Ski蛋白表达下调,TGF-β1/Smad3信号活化减弱,纤维化病变减轻。
Objective:To observe the expression changes of Ski protein in the kidney of diabetic mouse(DM) before and after the blood glucose control,and to explore its role in the pathogenesis of diabetic nephropathy(DN).Methods:Streptozotocin was used to replicate the DM mice model.24 SD mice were randomly assigned to 3 groups:normal group(NC),DM group and INS group.DM rats were induced by streptozotocin in DM group and INS group.INS group was treated with 22 U/(kg·D)insulin for 6 weeks after 4 weeks of successful modeling.The level of HbA1c,blood glucose and 24 h urine protein were determined with biochemical method.The pathological changes of renal tissue were observed.The expressions of Ski,Collagen-Ⅲ and E-cadherin were detected by immunohistochemical staining.The expression of Collagen-Ⅲ,Smad3 and p-Smad3(Ser432/425) in renal tissue of SD mice were detected by Western Blot.Results:Compared with the NC group,the mice in the DM group had reduced body weight,increased blood glucose and HbA1 c,and significant changes in renal tubular and glomerular fibrosis.The above changes were improved in the INS group.Immunohistochemical staining showed that the kidney tissue Ski and Collagen-Ⅲ expression from high to low were DM group>INS group>NC group(P<0.05).The expression of E-cadherin from high to low was NC group>INS group>DM group(P<0.05);The results of Western Blot showed that the expression of renal tissue TGF-β1,Collagen-Ⅲ,p-Smad3(Ser 432/425) and Ski protein from high to low were DM group>INS group>NC group(P<0.05).Conclusion:The expression of Ski protein is increased in the kidney of diabetic mice,and the activation of TGF-β1/Smad3 pathway leads to obvious fibrotic lesions.After insulin treatment,the expression of Ski protein is down-regulated;the signal transduction of TGF-β1/Smad3 weakened;the fibrotic lesions alleviated.

关键词(KeyWords)： 血糖;糖尿病肾病;肾纤维化;Ski蛋白;胰岛素;转化生长因子β1;大鼠,Sprague-Dawley
blood glucose;diabetic nephropathy;renal fibrosis;Ski protein;insulin;transforming growth factor β1;rats,Sprague-Dawley

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81760131);; 贵州省科技厅联合基金资助项目[黔科合LH字(2014)7124];; 贵州医科大学博士启动基金资助项目[院博合J字(2015)007号];; 贵阳市-贵州医科大学联合基金资助项目[筑科合同(20161001)]

作者(Author): 梁露群,任飞燕,刘玲伶,彭伟,张小欢,毛彦稳,刘慧铭,王圆圆,石明隽,郭兵
LIU Luqun,REN Feiyan,LIU Lingling,PENG Wei,ZHANG Xiaohuan,MAO Yanwen,LIU Huiming,WANG Yuanyuan,SHI Mingjun,GUO Bing

DOI: 10.19367/j.cnki.1000-2707.2019.01.002

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