基于TGF-β1/p38MAPK通路探讨益血降浊汤对UUO大鼠肾纤维化的作用机制

Study on the Mechanism of Yixue Jiangzhuo Decoction on Renal Fibrosis in UUO Rats Based on TGF-β1/p38MAPK Pathway

目的探讨益血降浊汤对单侧输尿管结扎(UUO)大鼠肾纤维化的作用及TGF-β1/p38MAPK通路相关蛋白的影响,阐明其作用可能的机制。方法建立UUO大鼠模型,将42只大鼠随机平均分为假手术组、UUO组、UUO+依那普利组、UUO+益血降浊汤低、中和高剂量组。于2周后处死,记录尿量、体重数据,采集血液检测氧化应激相关指标MDA、ROS和SOD含量。并取肾组织,Masson染色观察肾组织病理改变;免疫组织化学染色检测COX-2、Collagen I和p-p38MAPK表达情况;蛋白免疫印迹检测炎症、细胞外基质(ECM)以及TGFβ1/p38MAPK通路相关蛋白的表达。结果与UUO组相比,益血降浊汤(低、中、高)均能逆转大鼠尿量和体重的下降;降低ROS和MDA的水平,促进SOD的增加;减轻肾组织病理改变;下调炎症相关蛋白的表达,使抗炎因子的表达增加;减少ECM的沉积;抑制TGF-β1/P38MAPK通路活性;其中,高剂量组效果最为显著。结论益血降浊汤可能通过调控TGF-β1/p38MAPK通路活性改善炎症和氧化应激、抑制肾纤维化来发挥肾保护作用。

Objective:To study the effects of Yixue Jiangzhuo Decoction on renal fibrosis in rats with unilateral ureteral obstruction(UUO)and the related proteins of TGF-β1/p38MAPK pathway,and to elucidate its possible mechanism.Methods:A UUO rat model was established.42 rats were randomly divided into sham operation group,UUO group,UUO+enalapril group,and low,medium and high dose groups of UUO+Yixue Jiangzhuo Decoction.After 2 weeks,the rats were sacrificed,and their urine volume and body weight data were recorded and their blood was taken to detect the content of related indicators MDA,ROS and SOD for oxidative stress.Their renal tissue was taken to observe the pathological changes of renal tissues by Masson staining.The expressions of COX-2,Collagen I and p-p38MAPK were detected by immunohistochemical staining.Inflammation,extracellular matrix(ECM)and TGFβ1/p38MAPK pathway-related proteins were detected by western blotting.Results:Compared with the UUO group,Yixue Jiangzhuo Decoction(low,medium and high doses)can reverse the decrease of urine volume and body weight in rats,reduce the levels of ROS and MDA,promote the increase of SOD and relieve the pathological changes of renal tissues.It also down-regulated the expression of inflammation-related proteins,increased the expression of anti-inflammatory factors,decreased the deposition of ECM and inhibited the activity of TGF-β1/P38MAPK pathway.Among them,the high-dose group was the most effective.Conclusion:Yixue Jiangzhuo Decoction may play a role in renal protection by regulating the activity of TGF-β1/p38MAPK pathway to improve inflammation and oxidative stress and inhibit renal fibrosis.