吡非尼酮对大鼠肾间质纤维化的防治作用研究

Effect of Pirfenidone on renal interstitial fibrosis in rats

摘要:

目的研究吡非尼酮（PFD）对大鼠肾间质纤维化的作用及其作用机制，为临床使用吡非尼酮提供理论依据。方法24 只标准雄性SD 大鼠，随机分为3 组：正常组（NC 组）、模型组（M 组）及吡非尼酮治疗组（S组）。M组及S 组行左侧输尿管结扎术，复制肾间质纤维化模型。S 组予吡非尼酮250 mg/（kg·d）灌胃，NC 组及M 组大鼠予等量1%羧甲基纤维素钠溶液灌胃。分别于手术后第1、3、5、7 天处死各组大鼠各2 只，取梗阻侧肾脏。HE 染色观察各组肾组织病理变化，实时荧光定量聚合酶链反应（qRT-PCR）检测结缔组织生长因子（CTGF）mRNA 表达水平，Western blot 检测转化生长因子-β（TGF-β）及α- 平滑肌激动蛋白（α-SMA）的蛋白表达水平。结果NC 组未显示肾纤维化，M 组显示大鼠肾间质纤维化及炎症细胞浸润。NC 组TGF-β、α-SMA 蛋白表达均较低，M 组TGF-β、α-SMA 蛋白及CTGF mRNA 表达较NC 组均升高，差异有统计学意义（p <0.05），且随着梗阻时间延长，各物质表达量增高，但第7 天M 组及S 组上述物质表达量较第5 天减少，且M 组与S 组之间表达差异无统计学意义（p >0.05）；S 组上述物质各时间点表达则较M 组均减少，差异有统计学意义（p <0.05）。结论吡非尼酮可能通过抑制大鼠肾组织TGF-β和CTGF 的表达进而减少成纤维细胞转化为肌成纤维细胞，减少细胞外基质（ECM）的沉积，发挥抗肾纤维化作用。

Abstract:

Objective To investigate the effect of Pirfenidone (PFD) on renal interstitial fibrosis in rat model of unilateral ureteral occlusion (UUO) and to reveal the anti -fibrosis mechanism of PFD. Methods Twenty -four standard male SD rats were randomly divided into three groups: normal control group(N), UUO model group (M) and Pirfenidone treated group (S). The rats of the M and S groups underwent left urethral ligation to make the renal interstitial fibrosis model. The S group was given PFD 250 mg/(kg·d) for gavage, meanwhile the N and M groups were given 1% sodium carboxymethyl cellulose solution for gavage. Two rats were sacrificed from each group on day 1, 3, 5 and 7 after UUO operation, the kidney of the obstructive side was taken for further research. In each group,pathological changes were observed by HE staining, qRT-PCR was applied to detect the mRNA expression of connective tissue growth factor (CTGF), Western bolt was applied to detect the protein expressions of TGF-βand α-SMA. Results There was obvious renal interstitial fibrosis in the M group. Compared with the N group, the expressions of TGF-βand α-SMA proteins and the mRNA expression of CTGF were significantly increased in the M group(p < 0.05). Compared with the M group, the expressions of above parameters in the S group were decreased significantly(p < 0.05). The level of TGF -βwas positively correlated with the level of α-SMA (p < 0.05). Conclusions Pirfenidone has anti -fibrosis effect on rat UUO model by inhibiting renal TGF -β and CTGF expressions, which may control the fibroblast transformation and reduce the extracellular matrix deposition in the renal tissue.