miR-122在糖尿病肾病小鼠肾组织中表达变化及作用机制  

Expression and mechanism of miR-122 actions in renal tissue of diabetic nephropathy mice

作　　者：樊东哲[1] 张晓斌[1] 牟淑敏[2] FAN Dongzhe;ZHANG Xiaobin;MU Shumin(Department of Nephrology Center,the Second Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250000,China;Department of Endocrine,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250000)

机构地区：[1]山东中医药大学第二附属医院肾病诊疗中心,济南250000 [2]山东中医药大学附属医院内分泌科,济南250000

出　　处：《中国比较医学杂志》2021年第10期36-41,共6页Chinese Journal of Comparative Medicine

基　　金：山东省中医药科技发展计划项目(2019-0083)。

摘　　要：目的探讨miR-122在糖尿病肾病(diabetic nephropathy,DN)小鼠肾组织中的表达变化以及沉默miR-122对DN小鼠肾组织的影响及可能的机制。方法采用链脲佐菌素(STZ)腹腔注射联合高糖高脂饮食建立C57BL/6小鼠DN模型。将造模成功的30只DN小鼠随机分为模型组、antagomir-NC组、antagomir-122组,每组10只,另外选取未做任何处理的健康C57BL/6小鼠10只作为正常对照组。造模成功后,antagomir-122组和antagomirNC组每7 d分别给予尾静脉注射antagomir-122和antagomir-NC进行干预,模型组和正常对照组给予尾静脉注射等量生理盐水代替。8周后收集血清、尿液后处死小鼠取得肾标本,自动生化仪检测血清肌酐(Cr)、尿素氮(BUN)和24 h尿蛋白定量水平,过碘酸雪夫(PAS)染色评价肾组织病理学变化,qRT-PCR检测肾组织miR-122的表达水平,试剂盒检测肾组织谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平,Western blot检测肾组织Sirt1、α-SMA、fibronectin蛋白的表达水平。结果与正常对照组比较,模型组小鼠肾小球硬化评分上升(P<0.05),肾组织miR-122和α-SMA、fibronectin蛋白的表达明显升高(P<0.05),血清Cr、BUN水平,24 h尿蛋白定量水平,肾组织MDA水平明显增高(P<0.05),肾组织GSH-Px和SOD水平、Sirt1蛋白表达水平均明显降低(P<0.05)。模型组与antagomir-NC比较,各项指标差异无统计学意义(P>0.05)。与antagomir-NC组比较,antagomir-122组小鼠肾小球硬化评分下降(P<0.05),肾组织miR-122和α-SMA、fibronectin蛋白的表达明显降低(P<0.05),血清Cr、BUN水平,24 h尿蛋白定量水平,肾组织MDA水平明显下降(P<0.05),肾组织GSH-Px和SOD水平、Sirt1蛋白表达水平均明显升高(P<0.05)。结论miR-122在DN小鼠肾组织中表达升高。沉默miR-122通过抗氧化应激和纤维化对DN小鼠肾组织起到明显的保护作用,其机制可能与其对Sirt1基因的调控有关。Objective To investigate the expression of mi R-122 in the kidneys of diabetic nephropathy(DN) mice and the effect of mi R-122 in the kidney of DN mice.Methods Streptozotocin(STZ)intraperitoneal injection combined with a high-sugar and high-fat diet was used to establish the DN C57 BL/6 mouse model.Thirty DN mice were randomly divided into the model group,antagomir-NC group and antagomir-122 group(10/group).In addition,10 healthy C57 BL/6 mice without any treatment were selected as a normal control group.After the successful establishment of the model,the antagomir-122 and antagomir-NC groups were given tail vein injections of antagmir-122 and antagomir-NC every7 days,and the model and normal control groups were given tail vein injections of the same volume of normal saline.After 8 weeks,serum and urine samples were collected and mice were sacrificed to obtain kidney specimens.An automatic biochemical analyzer was used to detect serum creatinine(Cr),blood urea nitrogen(BUN)and 24 h urine protein levels.Periodic acid schiff staining was used to evaluate renal histopathological changes,q RT-PCR was used to detect expression levels of mi R-122 in renal tissue,and a kit was used to detect levels of glutathione peroxidase(GSH-Px),malondialdehyde(MDA),and superoxide dismutase(SOD)in renal tissues.Western blot was used to detect protein levels of Sirtuin 1(Sirt1),α-smooth muscle actin(α-SMA),and fibronectin in renal tissue.Results Compared with the normal control group,the glomerulosclerosis score of mice increased(P<0.05),the expression of mi R-122,α-SMA and fibronectin protein in renal tissue increased(P<0.05),circulating levels of Cr and BUN,and quantitative levels of 24 h urine protein and MDA levels in renal tissue were all significantly increased(P<0.05),and the expression levels of Sirt1 protein,and SOD and GSH-Px levels in renal tissue were all significantly reduced(P<0.05)in the model group.There were no significant differences in the endpoints examined between the model group and antagomir-NC group.Compared

关 键 词：MIR-122 SIRT1 小鼠 糖尿病肾病 氧化应激 纤维化 

分 类 号：R-33[医药卫生]