1,25-（OH）2D3对阿霉素肾病模型大鼠足细胞损伤及podocin表达缺失的影响  

Effects of 1,25-（OH）2D3 on podocyte injury and loss of podocin expression in rats with adriamycin-induced nephropathy

作　　者：赵丹[1] 陈亚茹 于曼丽 马国英 杨晓萍[1] 罗星[2] ZHAO Dan;CHEN Ya-Ru;YU Man-Li(Department of Nephrology,the First Affiliated Hospital of Medical College of Shihezi University,Shihezi 832002,Xinjiang,China)

机构地区：[1]石河子大学医学院第一附属医院肾病科,新疆石河子832002 [2]石河子大学医学院生化教研室,新疆石河子832002

出　　处：《中国老年学杂志》2019年第13期3225-3230,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(81160090,31160182);石河子大学应用基础研究青年项目(2015ZRKXYQ-LH20);石河子大学大学生研究训练计划资助项目(201810759020)

摘　　要：目的研究活性维生素D3〔1,25-(OH)2D3〕对阿霉素(ADR)肾病模型大鼠足细胞的作用及对podocin表达的影响。方法90只雄性SD大鼠随机选取30只作为对照组,其余60只被一次性尾静脉注射ADR7.5mg/kg。建模成功后,随机均分为ADR组、治疗组,治疗组每日给予1,25-(OH)2D30.25μg/kg灌胃,对照组及ADR组每日给予等量的花生油灌胃。于治疗第2、4、6、8、10周末收集24h尿,进行生化分析;取肾组织进行后续的组织学观察和分子水平检测。结果与对照组相比,ADR组24h尿蛋白显著增加(P<0.05),1,25-(OH)2D3显著减少24h尿蛋白含量(P<0.05)。苏木素-伊红(HE)染色结果显示,对照组肾小球形态完好,ADR组肾小球基底膜增厚、肾间质纤维化、大量炎性细胞浸润,1,25-(OH)2D3治疗减轻了肾小球损伤。电镜观察显示:与对照组比较,模型组足细胞超微结构发生改变,出现细胞空腔、细胞核畸形,基底膜呈节段性增厚、足突被压扁且融合;1,25-(OH)2D3治疗后,足细胞损伤减轻。免疫组织化学检测显示:podocin主要定位于细胞膜,在正常肾小球中高表达,而ADR组表达较低(P<0.05);1,25-(OH)2D3治疗后,podocin的表达显著增高(P<0.05)。荧光定量PCR和Western印迹结果显示:podocin mRNA和蛋白水平具有高度的一致性,与对照组相比,ADR组表达显著降低(P<0.05);1,25-(OH)2D3治疗后,podocin表达有所恢复。结论1,25-(OH)2D3能够改善ADR肾病模型大鼠肾脏足细胞损伤,逆转podocin蛋白表达缺失,从而减少24h尿蛋白,发挥重要的肾脏保护作用。Objective To investigate the effect of active vitamin D3 〔active vitamin D3,1,25-(OH)2D3〕 on adriamycin (ADR) podocytes and its effects on podocin expression. Methods Thirty male SD rats were randomly selected as control group,60 rats were successfully modeled with 7.5 mg/kg ADR and divided into ADR group and treatment group (ADR+1,25-D3).Active vitamin D3 was given on the third day after the model in treatment group and the control group was given the same amount of peanut oil.After the injection of ADR 2,4,6,8,10 weeks,24 h urine samples were collected for biochemical analysis.The kidney tissue was taken for subsequent histological observation and molecular level detection. Results The biochemical results showed that 24 h urine protein group was significantly increased in model group than that of control group at the same period ( P <0.05),1,25-(OH)2D3 could significantly reduce 24 h urine protein content.Histochemical detection:in the control group,glomerular morphology was intact.Basement membrane thickening,renal interstitial fibrosis,and infiltration of a large number of inflammatory cells were observed in the glomeruli of the model group,and glomerular injury was alleviated after 1,25-(OH)2D3 treatment.Transmission electron microscopy results showed that compared with the control group,the ultrastructure of the podocyte of the model group changed,the cell cavity and nucleus malformation appeared,the basement membrane showed segmental thickening,the foot process was squashed and fusion in model group.The podocyte injury was decreased after 1,25-(OH)2D3 treatment,immunohistochemistry showed that podocin was mainly localized on the cell membrane and was highly expressed in normal glomeruli,whereas it was lower in the model group ( P <0.05),and podocin expression was significantly higher after 1,25-(OH)2D3 treatment ( P <0.05).The results of fluorescence quantitative PCR and Western blot showed that the expression level of podocin mRNA and protein were highly consistent.Compared with the control group,the

关 键 词：活性维生素D3 慢性肾脏病 足细胞 PODOCIN 

分 类 号：R692[医药卫生—泌尿科学]