上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (7): 752-.

• 论著（基础研究） • 上一篇    下一篇

UUO模型大鼠肾间质纤维化动态进展及α-SMA、TGF-β1和VDR表达变化

谢盛彬, 王伟铭, 陈 楠   

  1. 上海交通大学 医学院瑞金医院肾脏内科, 上海 200025
  • 出版日期:2010-07-25 发布日期:2010-07-26
  • 通讯作者: 陈 楠, 电子信箱: chen-nan@medmail.com.cn。
  • 作者简介:谢盛彬（1983—）, 男, 硕士生；电子信箱: smilore_0701@sina.com。
  • 基金资助:

    上海市科委基金（08dz1900502）；诺华科研基金；上海市重点学科基金（T0201）

Progression of renal tubulointerstitial fibrosis and expression of α-SMA, TGF-β1 and VDR in rat UUO models

XIE Sheng-bin, WANG Wei-ming, CHEN nan   

  1. Department of Nephrology, Ruijin Hospital, School of Medicine, |Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2010-07-25 Published:2010-07-26
  • Supported by:

    Shanghai Science and Technology Committee Foundation, 08dz1900502；Novartis Research Foundation；Shanghai Key Discipline Foundation,T0201

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摘要：

目的 观察单侧输尿管梗阻(UUO)模型大鼠肾小管间质纤维化动态进展，研究该病理过程中肾脏纤维化相关蛋白表达的变化。方法 32只清洁级SD大鼠随机分为假手术组（n=16）和UUO模型组(模型组，n=16)。两组大鼠分别于术后（模型组建模后）第2、5、9、14天分批（n=4）处死，留取手术侧（模型组梗阻侧）肾脏组织。分别采用HE和Masson染色、免疫组织化学染色及Western blotting等方法，评价肾小管间质纤维化损伤程度并检测肾脏组织α平滑肌肌动蛋白（α-SMA）、纤维连接蛋白（FN）、维生素D受体（VDR）及转化生长因子β1（TGF-β1）等相关蛋白的表达。结果 肾脏组织形态学观察显示，随着输尿管梗阻时间的延长，肾小管间质纤维化程度逐步加重。免疫组织化学染色显示，模型组大鼠建模后各时间点肾间质α-SMA和FN阳性面积百分比均显著高于假手术组（P<0.05），且随着梗阻时间的延长逐渐升高，至建模后第14天时达到高峰。Western blotting分析表明，模型组大鼠建模后各时间点α-SMA和TGF-β1相对表达量均显著高于假手术组（P<0.05），而VDR相对表达量显著低于假手术组（P<0.05）；建模后第2天，模型组大鼠肾脏组织α-SMA相对表达量已显著增加；建模后第14天，模型组α-SMA和TGF-β1相对表达量分别增高至假手术组的12.7倍和8.8倍，而VDR相对表达量下降至假手术组的3%。结论 UUO模型大鼠建模后第14天可出现显著的肾间质纤维化；建模早期肾间质中α-SMA表达增加提示肾间质成纤维细胞的活化；VDR表达的进行性减少提示其与肾间质纤维化有关。

关键词: 肾小管间质纤维化, 单侧输尿管梗阻, α平滑肌肌动蛋白, 维生素D受体, 转化生长因子β1

Abstract:

Objective To observe the progression of renal tubulointerstitial fibrosis in rat unilateral ureteral obstruction (UUO) models, and investigate the changes in expression of renal fibrosis-related proteins during the pathological course. Methods Thirty-two SD rats were randomly divided into sham-operation group (n=16) and UUO model group (model group, n=16). Rats were sacrificed 2, 5, 9 and 14 d after operation (after model establishment for model group) (n=4 for each day), and the renal tissues of the operated side (obstructed side for model group) were obtained. HE staining, Masson staining, immunohistochemical staining and Western blotting were used to evaluate the degree of renal tubulointerstitial fibrosis and detect the expression of α-smooth muscle actin (α-SMA), fibronectin (FN), vitamin D receptor (VDR) and transforming growth factor-β1 (TGF-β1) in renal tissues. Results Histological observations of renal tissues revealed that the degree of renal tubulointerstitial fibrosis gradually increased with time of ureteral obstruction. It was indicated by immunohistochemical staining that the percentages of area with positive α-SMA and FN in renal interstitium of model group, which gradually increased with time of ureteral obstruction and reached the peak 14 d after model establishment, were significantly higher than those of sham-operation group at each time point after model establishment (P<0.05). Western blotting analysis demonstrated that the relative expression of α-SMA and TGF-β1 at each time point after model establishment in model group was significantly higher than that in sham-operation group (P<0.05), while the relative expression of VDR was significantly lower than that in sham-operation group (P<0.05). The relative expression of α-SMA in renal tissues of rats in model group significantly increased on the second day after model establishment, and the relative expression of α-SMA and TGF-β1 in model group was 12.7 times and 8.8 times respectively of that in sham-operation group on the fourteenth day after model establishment, while the relative expression of VDR decreased to 3% of sham-operation group. Conclusion Significant interstitial fibrosis may emerge at 14 d after model establishment in rat UUO models.The expression of α-SMA in renal interstitium increases in the early stage of model establishment, indicating the activation of renal interstitial fibroblasts. The progression of decrease in expression of VDR indicates VDR may relate to renal interstitial fibrosis.

Key words: renal tubulointerstitial fibrosis, unilateral ureteral obstruction, α-smooth muscle actin, vitamin D receptor, transforming growth factor-β1