比较胰岛素样生长因子1受体抑制剂与胰岛素对2型糖尿病肾病小鼠肾间质巨噬细胞浸润的改善作用  

Comparation of the effects of insulin-like growth factor-1 receptor inhibitor and insulin on renal interstitial macrophage infiltration in mice with type 2 diabetic kidney disease

作　　者：陈爽[1] 全裕俊 董蓉[1] 俞佳丽 张茜[1] 贺蓉 袁静[1] 查艳[1] Chen Shuang;Quan Yujun;Dong Rong;Yu Jiali;Zhang Qian;He Rong;Yuan Jing;Zha Yan(Department of Nephrology,Guizhou Provincial People's Hospital,Guiyang 550002,China;Department of Nephrology,Jianhe County People's Hospital,Jianhe County of Guizhou Province 556400,China)

机构地区：[1]贵州省人民医院肾内科,贵阳550002 [2]贵州省剑河县人民医院肾内科,贵州省剑河县556400

出　　处：《中华肾脏病杂志》2019年第10期765-772,共8页Chinese Journal of Nephrology

基　　金：国家自然科学基金(81760134);贵州省高层次创新型人才(黔科合平台人才[2018]5636);贵州省科技合作计划项目(黔科合LH字[2014]7003、黔科合LH字[2015]7155).

摘　　要：目的比较胰岛素样生长因子1受体(insulin-like growth factor-1 receptor,IGF-1R)抑制剂与胰岛素对2型糖尿病肾病(DKD)小鼠肾间质巨噬细胞浸润的改善作用。方法选取24只雄性C57BL/6小鼠,适应性喂养1周后,随机抽取6只作为对照组;其余小鼠高脂高糖喂养8周后,腹腔注射链脲佐菌素(30mg/kg),72h后随机血糖大于16.7mmol/L,为2型糖尿病(DM)模型建立成功。DM小鼠8周后出现尿蛋白增加,即DKD造模成功。采用随机数余数法将DKD小鼠分为3组:DKD组、DKD+胰岛素组(胰岛素组,皮下注射1~2U/d胰岛素)和DKD+IGF-1R抑制剂组(IGF-1R抑制剂组,给予30mg·kg^-1·d^-1 IGF-1R抑制剂灌胃),每组6只,持续干预8周。血糖仪检测血糖;收集血液和尿液,生化仪检测血肌酐、尿素氮和尿蛋白等生化指标。收集肾脏组织,用苏木精-伊红染色(HE)和过碘酸希夫染色(PAS)检测肾脏病理变化,原位杂交检测细胞因子信号转导抑制因子2(suppressor of cytokine signaling2,SOCS2)和胰岛素样生长因子1(insulin-like growth factor-1,IGF-1)mRNA,免疫组化检测SOCS2、F4/80、Toll样受体4(TLR4)、CD68蛋白的表达。结果相对于对照组,DKD小鼠血糖、血肌酐、血清尿素氮和尿蛋白排泄率明显较高(均P<0.05),且DKD小鼠肾小管间质中CD68^+细胞、F4/80^+细胞以及TLR4表达均明显较多(均P<0.05)。经胰岛素或IGF-1R抑制剂干预后,DKD小鼠血肌酐、血清尿素氮和尿蛋白排泄率均明显减少(均P<0.05)。胰岛素干预可以明显降低小鼠血糖(P<0.05),但对巨噬细胞无明显影响。IGF-1R抑制剂虽没有明显降低血糖,但是可明显降低DKD小鼠肾间质中CD68、F4/80阳性细胞数及TLR4蛋白的表达(均P<0.05)。相对于DKD组,胰岛素干预明显减少IGF-1蛋白和mRNA表达(均P<0.01),增加SOCS2mRNA和蛋白表达(均P<0.01),且胰岛素组SOCS2蛋白表达与F4/80^+细胞数具有相关性(R^2=0.8461,P=0.005)。然而IGF-1R抑制剂组SOCS2表达无明显变化,但具有更好的抑制Objective To compare the effect of insulin-like growth factor-1 receptor (IGF-1R) inhibitor and insulin on renal interstitial macrophage infiltration in mice with type 2 diabetic kidney disease (DKD) mice. Methods Twenty-four male C57BL/6 mice were selected. After 1 week of adaptive feeding, 6 rats were randomly selected as the control group. The other mice were intraperitoneally injected with streptozotocin (30 mg/kg) after 8 weeks of high-fat and high-sugar feeding. After 72 h, the type 2 diabetes mellitus (DM) models were successfully established if random blood glucose was greater than 16.7 mmol/L. After 8 weeks, if the proteinuria of DM mice increased, the DKD models were successful. DKD mice were divided into 3 groups by random number remainder method: DKD group (n=6), DKD+insulin group (insulin group, n=6, subcutaneous injection of 1-2 U/d insulin) and DKD+IGF-1R inhibitor (IGF-1R inhibitor group, n=6, administered with 30 mg·kg^-1·d^-1 IGF-1R inhibitor). They were continuously treated for 8 weeks. Random blood glucose was tested by glucometer. Blood and urine were collected, and biochemical indicators, such as serum creatinine, urea nitrogen and urine protein were measured by biochemical analyzer. Renal pathological changes were detected by hematoxylin-eosin staining (HE) and periodic acid-schiff staining (PAS). Suppressor of cytokine signaling 2 (SOCS2) mRNA and insulin-like growth factor-1 (IGF-1) mRNA were detected by in situ hybridization. The protein expressions of SOCS2, F4/80, Toll-like receptor 4 (TLR4) and CD68 were detected by immunohistochemistry. Results Compared with the control group, blood glucose, serum creatinine, serum urea nitrogen and urinary protein excretion rate were significantly higher in DKD mice (all P<0.05), and CD68^+ cells number, F4/80^+ cells number and the expression of TLR4 in the tubulointerstitial of DKD mice were significantly higher (all P<0.05). After intervention with insulin or IGF-1R inhibitor, serum creatinine, serum urea nitrogen and urinary protein excretion

关 键 词：糖尿病肾病 胰岛素样生长因子Ⅰ 胰岛素 细胞因子信号转导抑制因子2 

分 类 号：R692[医药卫生—泌尿科学] R587[医药卫生—外科学]