姜黄素对单侧输尿管梗阻后肾间质纤维化的调节.doc

姜黄素对单侧输尿管梗阻后肾间质纤维化的调节 作者：张彬 王全胜 朱锐 汪永辉 张丽晓 徐敏 刘建国【摘要】 目的： 研究姜黄素在单侧输尿管梗阻(UUO) 大鼠模型中能否减少肾间质中转化生长因子1（transforming growth factor-1,TGF-1）和结缔组织生长因子（connective tissue growth factor，CTGF）的表达。 方法: 将30只大鼠随机分为3组，每组10只：假手术组、模型组、姜黄素组。模型组和姜黄素组行右侧输尿管接扎术，假手术组只游离不接扎。术后第14天处死各组中的大鼠，检测血液尿素氮（BUN）、血清肌酸酐（Scr）。用免疫组织化学方法测定TGF-1、CTGF的表达情况，同时采用Masson染色，观察肾脏病理情况。结果: 姜黄素降低了BUN、Scr的含量，同时姜黄素显著减少了大鼠肾间质TGF-1、CTGF的表达，并有效改善了肾脏的病理学损伤。 结论: 姜黄素可以减少输尿管梗阻后TGF-1、CTGF的表达，从而改善UUO所致的肾脏损伤，提示姜黄素可能有延缓慢性肾脏疾病纤维化进程的潜能。 【关键词】 输尿管梗阻； 纤维化； 姜黄素； 免疫组织化学； 大鼠，Spragua-Dawley； 肾间质 Abstract Objective: To verify if curcumin can decrease the expression of transforming growth factor-1(TGF-1) and connective tissue growth factor (CTGF) in unilateral ureteral obstruction (UUO)rat models during the process of renal interstitial fibrosis caused by unilateral ureteral obstruction (UUO).Methods: The experiment was successfully conducted from May to August, 2007 in Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology. Thirty Sprague-Dawley rats were randomly divided into three groups: sham operated group，UUO model group, and curcumin-treated group (10 in each group). The rats in the UUO model group and curcumin-treated group underwent UUO surgery. Rats in curcumin-treated group were given water suspension of curcumin powder (200 mgkg -1d -1) orally, while those in UUO model group and sham operation group were given water only instead. All of the rats were killed in 14 days after surgery. The levels of blood urea nitrogen (BUN) and serum creatinine (Scr) were measured, histological changes were observed with Masson stain method, and Immunohistochemistry was performed on renal tissue to detect TGF-1 and CTGF. Results: In the obstructed kidneys，there were significant renal interstieial fibrosis and interstitial space expansion. Immunohistochemical staining showed that the expression of TGF1 and CTGF was significantly increased in the obstructed kidneys. In comparison with the model group，BUN and Scr levels, interstitial volume, and the expression of TGF-1 and CTGF in curcumin-treated group were significantly decreased. Conclusions: The results indicate that curcumin can down-regulate the expression of TGF-1 and CTGF in UUO rats, through which curcumin might mitigate the pathological changes of the UUO renal tissue. Its suggested that curcumin can be used to slow down the progression of chronic renal disease. Key words ureteral obstruction; fibrosis; curcumin; immunohistochemistry; rats,Sprague-Dawley; renal inerestitum 肾间质纤维化几乎是所有慢性肾脏疾病进行性发展的共同最终通路，延缓肾间质纤维化的发展，可降低终末期肾病(ESRD)的发生率1。姜黄素是姜黄中提取的一种植物多酚，也是姜黄发挥药理作用最重要的活性成分。近年来的研究不仅证明了姜黄的传统作用，而且还揭示出一些新的药理作用，如抗炎、抗氧化、清除氧自由基、抗人类免疫缺陷病毒、保护肝脏和肾脏、抗纤维化以及防癌抗癌等作用，而且无明显的毒副作用2。2007年58月研究了姜黄素抑制肾间质纤维化的作用机制，其中重在研究姜黄素对在单侧输尿管梗阻（UUO）模型大鼠中肾间质中转化生长因子1（transforming growth factor-1,TGF-1）、结缔组织生长因子（connective tissue growth factor，CTGF）的影响，希望在临床上研制出有效药物。 1 材料与方法 1.1 材料 雄性、无特定病原体动物级（specific pathogen free, SPF）、Sprague-Dawley大鼠同济医学院动物中心提供，生产许可证：SCXK（鄂）2004-002830只，7 w龄，体重(180.28.35)g。动物房饲养环境为每笼6只，每24 h换1次垫料，喂以正常饮食，自由进水，温度为（242），相对湿度为（453）%。姜黄素纯度为95%（美国Sigma公司）；兔抗大鼠TGF-1、CTGF 多克隆抗体及SP免疫试剂盒（北京中杉金桥生物技术有限公司）。 1.2 方法 1.2.1 造模 适应性饲养1周后，按照完全随机设计方法，以体重由轻到重的顺序编号用随机排列表法分为假手术组、模型组、姜黄素组各10只。假手术组和模型组每天给予蒸馏水2 ml灌胃，姜黄素组将姜黄素粉末（200 mgkg-1d-1）混悬于2 ml蒸馏水中灌胃，术前1 d开始给药。造模方法：模型组和姜黄素组行右侧输尿管结扎术，质量浓度为20