摘要
目的探究微小RNA(miR)-194-5p通过靶向调节CD2相关蛋白(CD2AP)参与肾病综合征(NS)发展的机制。方法通过工具网站starbase预测miR-194-5p靶向CD2AP的潜在结合位点,然后通过荧光素酶报告验证。将大鼠分为3组(n=8):对照组、NS组和antagomir组,NS组和antagomir组通过静脉注射阿霉素建立NS模型。Antagomir组通过静脉注射miR-194-5p antagomir干预,其余两组注射等量溶剂。比较各组大鼠一般情况、24 h尿蛋白、肌酐水平。通过苏木精-伊红(HE)染色检测肾组织损伤情况。RT-PCR和Western blot检测CD2AP mRNA和蛋白的水平。结果通过双荧光霉素实验结果验证了同时转染miR-194-5p mimic和CD2AP-wt的细胞的荧光酶素活性显著降低(P<0.05)。过表达miR-194-5p显著抑制CD2AP mRNA和蛋白的表达(P<0.05)。NS组大鼠在给药后出现精神萎靡、脱毛、活动力下降等现象,在第2周后开始大鼠出现浮肿,并有竖毛现象。Antagomir组大鼠在给予miR-194-5p antagomir后,各种症状均有所缓解。建模后,NS组和antagomir组大鼠24 h尿蛋白和肌酐水平显著高于对照组,干预后antagomir组的24 h尿蛋白和肌酐均显著降低,并显著低于NS组(P<0.05)。NS组肾小球硬化、毛细血管扩张以及肾小球系膜增厚,并可观察到肾小管空泡样变性以及蛋白管型;antagomir组的肾小球和肾小管基本正常,病变情况显著轻于NS组。与对照组比较,干预后NS组大鼠肾组织匀浆中的miR-194-5p显著升高而CD2AP mRNA和蛋白显著降低(P<0.05)。而antagomir组的miR-194-5p显著低于NS组,CD2AP mRNA和蛋白显著高于NS组(P<0.05)。结论 miR-194-5p靶向抑制CD2AP的表达并参与NS的发病机制,抑制miR-194-5p可促进CD2AP的表达水平,从而减轻NS肾损伤。
Objective To explore the mechanism by which microRNA(miR)-194-5 p participates in the development of nephrotic syndrome(NS)by targeting CD2-associated protein(CD2 AP).Methods The potential binding site of miR-194-5 p targeting CD2 AP was predicted by the tool website starbase and then verified by luciferase reporter.Rats were divided into 3 groups(n=8),control group,NS group and antagomir group.The NS model was established by intravenous injection of doxorubicin.The antagomir group was injected with the same amount of solvent by intravenous injection of miR-194-5 p antagomir as comparison.The general condition,24 h urine protein and creatinine levels of each group were compared.Renal tissue damage was detected by Hematoxylin-eosin(HE)staining.QPCR and Western blot were used to detect mRNA and protein levels.Results The results of the dual fluorescein assay demonstrated that the luciferase activity of cells transfected with miR-194-5 p mimic and CD2 AP-wt was significantly decreased(P<0.05).Overexpression of miR-194-5 p significantly inhibited the expression of CD2 AP mRNA and protein(P<0.05).Rats in the NS group showed signs of wilting,hair loss,and decreased activity after administration.After the second week,the rats began to have edema and vertical hairiness.In the Antagomir group,various symptoms were alleviated after administration of miR-194-5 p antagomir.After modeling,the 24 h urine protein and creatinine levels in the NS group and the antagomir group were significantly higher than those in the control group.After intervention,the 24-hour urine protein and creatinine in the antagomir group were significantly lower than those in the NS group(P<0.05).In the NS group,glomerular sclerosis,telangiectasia,and mesangial thickening were observed,and tubular vacuolar degeneration and protein casts were observed.The glomeruli and renal tubules in the antagomir group were basically normal,and the lesions were significantly lighter than those in the NS group.Compared with the control group,the miR-194-5 p in the rena
作者
孙景毅
赵楠
潘红梅
张浩然
SUN Jing-yi;ZHAO Nan;PAN Hong-mei(Department of Nephrology,Hebei Cangzhou Central Hospital,Cangzhou Hebei 061000,China;Department of Nephropathy,Hebei Cangzhou Chinese and Western Medical Association Hospital,Cangzhou Hebei 061000,China)
出处
《临床和实验医学杂志》
2020年第6期595-599,共5页
Journal of Clinical and Experimental Medicine
