双环醇对单侧输尿管梗阻大鼠肾间质结缔组织生长因子、基质金属蛋白酶组织抑制物-1的影响

摘要

目的 观察双环醇对单侧输尿管梗阻(UUO)大鼠肾间质结缔组织生长因子(CTGF)、基质金属蛋白酶组织抑制物-1(TIMP-1)介导的肾间质纤维化(RIF)的影响,探讨其可能的肾脏保护机制.方法采用UUO致RIF大鼠模型.将大鼠随机分为假手术组、模型组、治疗组.从造模当天起,治疗组给予双环醇200 mg/(kg·d)灌胃,假手术组和模型组给予等体积生理盐水灌胃.术后第7、14、21天各组随机处死9只大鼠,肉眼观察肾脏的大体结构改变;行苏木精-伊红和Masson染色观察肾间质的病理学改变,评定小管间质损伤及肾间质纤维化程度;用免疫组织化学方法检测CTGF和TIMP-1的表达部位及蛋白表达水平;应用逆转录-聚合酶链反应(RT-PCR)检测肾组织TIMP-1 mRNA的表达水平.结果 假手术组大鼠肾脏大体及病理无明显异常,与假手术组比较,随着输尿管梗阻时限延长,模型组大鼠结扎侧肾脏明显肿大,肾间质损伤程度明显加重(P＜0.01),肾组织内CTGF、TIMP-1及TIMP-1 mRNA的表达均显著上调(P＜0.01).治疗组大鼠上述变化均较模型组轻(P＜0.01).结论 双环醇可能通过抑制TIMP-1、CTGF在UUO大鼠肾组织中的表达减轻UUO大鼠模型中肾间质纤维化的程度,发挥其对肾脏的保护作用.%Objective To observe the effect of bicyclol on renal interstitial expression of connective tissue growth factor (CTCF) and tissue inhibitor inelalloproleinase-l (TIMP-1) in rats with renal interstitial fibrosis induced by unilateral ureteral obstruction ( UUO) , and to explore the experimental evidence and theoretical basis for clinical trial of bicyclol on the renal interstitial fibrosis. Methods Renal interstitial fibrosis model was established via UUO. Tine rats were randomly divided into sham-operated group, UUO model group and bieyclol-treated group. After operations, the bicyclol-treated group was intragastncally administered with bicyclol I 200 mg/kg) until rats were killed. Sham-operated group and UUO model group were intragastncally administered with identical voluminal normal saline. In each group, nine rats were chosen randomly to be killed at the 7 d, 14 d and 21 d after operation. The gross structure of the kidneys was observed by naked eye and the renal tubulointerstitial damage and interstitial fibrosis were examined by routine hematoxylm and eosin (HE) and Masson staining under microscope. The expression of TIMP-1 and CTGF were detected by immuno-histochemical staining. The expression of TIMP-1-mRNA in kidney tissue was semi-quantitatively determined with reverse transcnption-polymcrasc chain reaction ( RT-PCR) . Results No anatomic or pathological change was observed on the kidneys of sham operated group. As compared with sham operated group, the kidneys on the iigated side were markedly enlarged, the renal tubulointerstitial lesions were more severe in UUO model group (P < 0.01). The renal tubulo interstitial expressions of CTCF, TIMP 1 and TIMP 1 mRNA were significantly up regulated (P < 0.01) in UUO model group. The severity of pathological changes depended on the duration of ureteral obstruction.These changes were lessened in rals of bicyclol-treated group (P < 0.01). Conclusions Bicyclol can alleviate the degree of renal interstitial fibrosis and protect kidney of UUO rats, probably by down--regulating the expression of TIMP-1 and CV,V.