氟非尼酮对单侧输尿管梗阻大鼠肾间质纤维化的疗效观察

Effect of fluorofenidone on renal interstitial fibrosis in rats with unilateral ureteral obstruction

目的:观察氟非尼酮对单侧输尿管梗阻大鼠肾间质纤维化的疗效,探讨氟非尼酮对肾间质纤维化大鼠肾组织中I型胶原(collagen type I,Col I),III型胶原(collagen type III,Col III),α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA),结缔组织生长因子(connective tissue growth factor,CTGF)和血小板源性生长因子(platelet derived growth factor,PDGF)表达的影响。方法:将15只雄性SD大鼠随机分为假手术组、模型组和氟非尼酮组(n=5)。对模型组和氟非尼酮组大鼠行左侧输尿管结扎术建立单侧输尿管梗阻模型。氟非尼酮组从造模前24 h开始以氟非尼酮[125 mg/(kg.d)]溶于0.5%羧甲基纤维素钠(sodium carboxyl methyl cellulose,CMC-Na)中灌胃,模型组和假手术组以等体积0.5%CMC-Na灌胃。于术后14 d处死大鼠,取梗阻侧肾组织行HE和Masson染色观察各组大鼠肾组织病理改变;用免疫组织化学检测Col I,Col III,α-SMA,PDGF,CTGF的蛋白表达;用RT-PCR检测Col I,Col III,PDGF,CTGF m RNA表达。结果:模型组肾间质损伤指数、胶原相对面积评分及Col I,Col III m RNA和蛋白表达均较假手术组增高(P<0.05),氟非尼酮治疗后明显改善(P<0.05)。模型组肾组织α-SMA,PDGF,CTGF蛋白表达及PDGF,CTGF m RNA表达均较假手术组明显增加(P<0.05),氟非尼酮治疗后表达均明显减少(P<0.05)。结论:氟非尼酮(125 mg/kg.d)能够显著改善单侧输尿管梗阻大鼠肾间质纤维化,其作用机制与抑制肌成纤维细胞的活化及关键促纤维化因子PDGF和CTGF的表达有关。

Objective： To investigate the effect of fluorofenidone on renal interstitial fibrosis in rats with unilateral ureteral obstruction （UUO） and to observe the effect of fluorofenidone on the expressions of collagen type I （Col I）, collagen type III （Col III）, α-smooth muscle actin （α-SMA）, connective tissue growth factor （CTGF）, platelet derived growth factor （PDGF） in the renal tissues of UUO rats.Methods： Male Sprague-Dawley （SD） rats were randomly divided into a sham-operated group,a UUO group, and a flurofenidone group （n=5）. UUO model was induced by ligating the left ureter in rats. The rats were treated with 125 mg/（kg.d） fluorofenidone by gastric gavage in the fluorofenidone group at 24 h before the operation, and the rats were treated with the identical dose of 0.5% sodium carboxyl methyl cellulose （CMC-Na） in the other 2 groups. The rats were sacrificed at 14 days after UUO. Pathological changes of the renal tissue were observed by HE and Masson staining, the mRNA expressions of Col I, Col III, α-SMA, PDGF and CTGF were detected by real-time PCR, and the protein expressions of Col I, Col III, PDGF and CTGF were detected by immunohistochemical staining.Results： The renal interstitial damage index, relative collagen area and mRNA and protein expressions of Col I and Col III in the renal tissues of the rats in the UUO group significantly increased （P〈0.05）, and fluorofenidone could reduce these indexes （P〈0.05）. Compared with the sham-operated group, the protein expressions of α-SMA, PDGF, CTGF and the mRNA expressions of PDGF and CTGF in the renal tissues of the rats in the UUO group were increased, but fluorofenidone could decrease the protein expressions of α-SMA, PDGF, CTGF and the mRNA expressions of PDGF and CTGF （P〈0.05）.Conclusion： Fluorofenidone （125 mg/kg.d） could attenuate renal interstitial fibrosis through inhibition of fibroblast proliferation, myofibroblastic activation, PDGF and CTGF expression.