槲皮素对单侧输尿管梗阻大鼠肾脏转化生长因子β1和结缔组织生长因子表达的调节效应

Regulative effects of quercetin on transforming growth factor beta 1 and connective tissue growth factor in kidney tissue in rats with unilateral ureteral occlusion

目的:观察槲皮素对大鼠梗阻肾脏间质纤维化的程度以及肾组织中转化生长因子β1、结缔组织生长因子表达的影响。方法:实验于2004-03/2005-02在吉林大学第三医院研究所完成。Wistar大鼠36只,雌雄不拘,体质量220～300g。①采用随机数字法随机分为3组,每组12只。单侧输尿管梗阻组:参照文献制备单侧输尿管梗阻肾间质纤维化大鼠动物模型。槲皮素组:即单侧输尿管梗阻模型+槲皮素治疗组,于手术前1天开始用槲皮素,剂量100mg/kg,灌胃,1次/d,连续10d。假手术组:打开腹腔后分离左输尿管,不结扎输尿管即关闭腹腔。②3组大鼠于术后10d,麻醉处死,取术侧肾组织做肾脏病理检查,在光学显微镜下观察肾间质病变程度。应用免疫组织化学法和蛋白免疫印记分析法检测大鼠梗阻侧肾脏转化生长因子β1、结缔组织生长因子的表达部位及蛋白表达。结果:大鼠36只全部进入结果分析。①各组大鼠肾脏病理学改变:单侧输尿管梗阻引起大鼠梗阻侧肾脏的间质纤维化,经槲皮素治疗后能够明显减轻单侧输尿管梗阻导致减轻肾脏病理损害。②各组大鼠免疫组织化学结果:与假手术组比较,单侧输尿管梗阻组和槲皮素组转化生长因子β1、结缔组织生长因子染色阳性表达增多(P<0.01);槲皮素组转化生长因子β1、结缔组织生长因子染色阳性表达比单侧输尿管梗阻组明显降低[(7.41±0.93)%比(13.39±0.87)%,P<0.01;(7.26±0.54)%比(10.18±0.49)%,P<0.05]。③各组大鼠免疫印记分析结果:与假手术组大鼠相比,单侧输尿管梗阻10d,大鼠梗阻侧肾脏组织内转化生长因子β1、结缔组织生长因子水平均明显增加(P<0.01),槲皮素组转化生长因子β1、结缔组织生长因子相对表达丰度明显低于单侧输尿管梗阻组[(2.42±0.56)%比(5.50±0.87)%,(3.26±1.54)%比(6.18±1.49)%,P<0.05]。结论:槲皮素具有抗肾间质纤维化作用,可能与其下调单侧输尿管梗阻大鼠肾脏组织中转化生长因子β1和结缔组织生长因子的表达有关。

AIM： To study the effects of quercetin on renal interstitial fibrosis and expressions of transforming growth factor （TGF） β1 and connective tissue growth factor （CTGF） in nephridial tissues of rats. METHODS： The experiment was performed in the Department of Third Hospital of Jilin Unversity from March 2004 to February 2005. Thirty-six Wistar rats of both sexes weighting 220-300 g were selected. ①All rats were randomly divided into 3 groups with 12 rats in each group. Unilateral ureteral occlusion （UUO） group： UUO rat models of renal interstitial fibrosis were established according to the references. Quercetin group i.e. UUO model ＋ quercetin treatment group; Rats were administrated with quercetin at one day before operation at the dose of 100 mg/kg once a day for 10 continuous days. Rats in the sham-operation group were separated of the left ureter after abdomen-opening without deligating the ureter. ② Rats were executed under anesthetization at 10 days after operation, and the nephridial tissues of operating side were detected. The pathological changes of renal interstitium were studied under the light microscope, lmmunohistochemical method and Western blot analysis were adopted to detect the expressions of TGF-β1 and CTGF in obstructed kidney of rats. RESULTS： A total of 36 rats were involved in the analysis of results.① Pathological changes of kidney in rats of each group： The renal interstitial fibrosis induced by UUO was obviously ameliorated after treatment with quercetin.②lmmunohistochemical results of rats in each group： compared with the sham-operation group, the stained positive expressions of TGF-β1 and CTGF in the UUO group and quercetin group were significantly increased （P 〈 0.01）, and those in the quercetin group were obviously decreased than those in the UUO group [（7.41±0.93）% vs. （13.39±0.87）%, P 〈 0.01; （7.26±0.54）% vs. （10.18±0.49）% ,P 〈 0.05];③Results of Western blot analysis in each group： compared with the sham-operation group, levels of TGF-β1 and CTGF in obstructed kidneys of rats on the 10th day of UUO were greatly increased （P 〈 0.01）, and the relative expression amount of TGF-β1 and CTGF in the quercetin group were obviously lower than the UUO group [（2.42±0.56）% vs. （5.50±0.87）%, （3.26±1.54）% vs.（6.18±1.49）%,P 〈 0.05]. CONCLUSION： Quercetin has anti-effect on renal interstitial fibrosis, which may be related with its down-regulation of TGF-β1 and CTGF in kidneys of rats with UUO.