上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (5): 567-.doi: 10.3969/j.issn.1674-8115.2011.05.009

• 论著（基础研究） • 上一篇    下一篇

AngⅡ诱导的肾损伤大鼠肾脏组织中SSeCKS的表达研究

王 涛, 王应灯, 张 薇   

  1. 上海交通大学 医学院附属第九人民医院肾脏内科, 上海 200011
  • 出版日期:2011-05-28 发布日期:2011-05-27
  • 通讯作者: 王应灯, 电子信箱: Wangyd7001@sina.com；张 薇, 电子信箱: Jiuyuan5139@yahoo.com.cn。
  • 作者简介:王 涛（1984—）, 男, 硕士生；电子信箱: wangtaonanyi@126.com。
  • 基金资助:

    上海市自然科学基金（09ZR1417400）

Expression of SSeCKS in renal tissues of rats with renal injury induced by AngⅡ

WANG Tao, WANG Ying-deng, ZHANG Wei   

  1. Department of Nephrology, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2011-05-28 Published:2011-05-27
  • Supported by:

    Shanghai Natural Science Foundation, 09ZR1417400

PDF (PC)

1733

摘要：

目的 建立由血管紧张素Ⅱ（AngⅡ）诱导的肾损伤大鼠模型，观察受损肾脏组织中Src抑制的蛋白激酶C的底物（SSeCKS）的表达情况。方法 26只Wistar大鼠随机分为模型组（n=10）、假手术组（n=10）和正常对照组（n=6）。模型组和假手术组大鼠经皮下埋置药物缓释泵分别向体内灌注AngⅡ和等量生理盐水，释放速度均为200 ng/(kg·min)；正常对照组大鼠不予任何处理。各组大鼠均于埋泵后第3天收集24 h尿样后处死，留取肾皮质组织。采用磺基水杨酸法进行24 h尿蛋白定量检测；透射电子显微镜观察大鼠肾脏组织超微结构变化；免疫荧光染色法观察和分析SSeCKS在大鼠肾脏组织中的表达部位及荧光强度；采用RT-PCR技术检测各组大鼠肾脏组织中SSeCKS mRNA的表达。结果 模型组大鼠24 h尿蛋白定量明显高于假手术组和正常对照组（P<0.05）。透射电子显微镜观察发现模型组大鼠肾小球内皮细胞皱缩、剥脱。免疫荧光染色分析结果显示：SSeCKS表达主要定位于肾小球基膜和部分系膜细胞区，模型组大鼠肾脏组织荧光强度显著高于假手术组和正常对照组。模型组大鼠肾脏组织中SSeCKS mRNA 表达明显高于假手术组和正常对照组。结论 通过皮下埋置AngⅡ药物缓释泵成功制作大鼠肾损伤模型，受损肾脏组织中SSeCKS的表达明显上调。

关键词: 肾损伤, 血管紧张素Ⅱ, Src抑制的蛋白激酶C的底物

Abstract:

Objective To establish the rat models of renal injury induced by angiotensionⅡ (AngⅡ), and investigate the expression of Src-suppressed C kinase substrate (SSeCKS) in the injured renal tissues. Methods A total of 26 rats were randomly divided into model group (n=10), sham operation group (n=10) and normal control group (n=6). Rats in model group and sham operation group were administered with AngⅡ and normal saline respectively via subcutaneous osmotic minipumps at the velocity of 200 ng/(kg·min), and those in control group did not receive any treatment. Twenty-four hour urine samples were collected on the third day after minipump implant, then rats were sacrificed, and renal cortical tissues were obtained. Sulfosalicylic acid method was adopted to quantify 24 h urinary protein, the ultrastructure of renal tissues were observed by transmission electron microscopy, the location of SSeCKS in renal tissues and fluorescence intensity were determined and analysed by immunofluorescence staining method, and the expression of SSeCKS mRNA in renal tissues was detected by RT-PCR. Results The quantity of 24 h urinary protein in model group was significantly higher than those in sham operation group and normal control group (P<0.05). Shrinkage and detachment of glomerular endothelial cells in model group were observed by transmission electron microscopy. Immunofluorescence staining revealed that SSeCKS mainly localized within glomerular basement and some mesangial cells, and fluorescence intensity of renal tissues in model group was significantly higher than that in sham operation group and normal control group. The expression of SSeCKS mRNA in renal tissues in model group was significantly higher than that in sham operation group and normal control group. Conclusion Rat models of renal injury have been successfully established by subcutaneous implant of osmotic minipump with AngⅡ, and the expression of SSeCKS may be significantly increased in the injured renal tissues.

Key words: renal injury, angiotensionⅡ, Src-suppressed C kinase substrateSrc