MiR-146a-5p对高脂饮食/链脲佐菌素诱导的糖尿病肾病大鼠肾组织的保护作用  

Protective effect of miR-146a-5p on renal tissue in high-fat diet/streptozotocin-induced diabetic nephropathy rats

作　　者：李莉 李松 曹萌[2] 石小娟 王雪 王顺阁 Li Li;Li Song;Cao Meng;Shi Xiaojuan;Wang Xue;Wang Shunge(Department of Endocrinology,Pingdingshan Second People's Hospital,Pingdingshan 467000;Department of Endocrinology,the First Affiliated Hospital of Xinxiang Medical College,Weihui 453100,China)

机构地区：[1]平顶山市第二人民医院内分泌科,平顶山467000 [2]新乡医学院第一附属医院内分泌科,卫辉453100

出　　处：《解剖学杂志》2021年第2期108-113,共6页Chinese Journal of Anatomy

基　　金：河南省医学科技攻关计划项目(201702183)。

摘　　要：目的:探究miR-146a-5p对高脂饮食(HFD)和链脲佐菌素(STZ)诱导的糖尿病肾病(DN)大鼠肾的保护作用及其作用机制。方法:SD大鼠随机分为正常对照组、模型组、miR-146a-5p阴性对照组(mimic对照组)和miR-146a-5p mimic处理组(mimic处理组)。测量各组大鼠体质量及最后24 h饮水量和尿量;全自动生化分析仪检测尿液中尿蛋白,血液中血糖、血脂、尿素氮和血清肌酐含量;H-E染色观察肾小球损伤;TUNEL染色观察肾组织凋亡,免疫印迹检测肾组织中凋亡标记物半胱氨酸蛋白酶3(caspase 3,cas3),淋巴细胞瘤-2相关蛋白X(Bax)和B淋巴细胞瘤-2(Bcl-2)的表达;酶联免疫吸附测定(ELISA)检测外周血中炎症因子肿瘤坏死因子-α(TNF-α),诱导型一氧化氮合酶(iNOS)、白细胞介素(IL)-6(IL-6)和IL-10的含量,免疫印迹检测肾组织中iNOS和IL-10的蛋白表达。结果:与正常对照组相比,HFD/STZ诱导的模型组大鼠体质量明显降低,饮水量、尿量和血液生化指标显著升高;肾小球体积增大,基底膜明显扩张,伴有大量炎性细胞浸润,且细胞凋亡数目明显增多,Bax/Bcl-2比值和cleaved cas3表达显著上调;炎症因子TNF-α、iNOS、IL-6和IL-10的水平明显上调。与模型组相比,mimic处理组大鼠体质量明显增加,饮水量、尿量及血液生化指标显著降低;肾小球体积增大,基底膜明显扩张及炎性细胞的浸润等病理变化明显缓解,细胞凋亡数显著减少,Bax/Bcl-2比值和cleaved cas3表达显著下调;促炎因子TNF-α、iNOS和IL-6的水平明显降低,抗炎因子IL-10的水平明显升高,mimic对照组差异无统计学意义。结论:MiR-146a-5p能减轻HFD/STZ诱导的糖尿病肾病大鼠肾损伤,其机制是通过抑制肾细胞凋亡及炎症反应,阻止糖尿病肾病的进一步发展。Objective:To explore the protective effect of miR-146a-5p on kidneys of high-fat diet(HFD)and streptozotocin(STZ)-induced diabetic nephropathy(DN)rats and its mechanism.Methods:Forty SD rats were randomly divided into 4 groups:control group(Control),model group(DN),miR-146a-5p negative control treatment group(DN+mimic mock)and miR-146a-5p mimic treatment group(DN+mimic).DN rats were induced by high-fat diet and 35 mg/kg streptozotocin(HFD/STZ).DN rats in the treatment group was injected with the lentiviral-coated miR-146a-5p mimic mock and miR-146a-5p mimic sequences through the tail vein,twice a week for 8 weeks,the same amount of normal saline was injected at the same time in rats of both the control group and the DN group.After 56 days of treatment,the weight of each group of rats and the drinking water and urine volume of the last 24 hours were measured;the automatic biochemical analyzer was used to detect urine protein,blood glucose,blood lipids,urea nitrogen and serum creatinine.H-E staining was employed to observe glomerular injury.TUNEL staining was adopted to observe the apoptosis of renal tissues.Western blotting was performed to detect the expression of apoptosis markers caspase 3(cas3),Bax,and Bcl-2 in renal tissues.ELISA was used to detect the content of inflammatory factors TNF-α,iNOS,IL-6 and IL-10 in peripheral blood.Western blotting was employed to detect expression of iNOS and IL-10 protein in renal tissues.Results:Compared with the control group,the body mass of DN rats induced by HFD/STZ was significantly reduced,and the water intake,urine output and blood biochemical indicators were significantly increased.The glomerular volume increased and the basement membrane expanded significantly,with a large number of infiltration of inflammatory cells,and the number of apoptosis increased significantly.Bax/Bcl-2 ratio and the expression of cleaved cas3 were significantly upregulated.The levels of inflammatory factors TNF-α,iNOS,IL-6 and IL-10 were significantly increased.Compared with the DN group,b

关 键 词：糖尿病肾病 miR-146a-5p 高脂饮食 链脲佐菌素 炎症因子 凋亡 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]