诃子酸抗肾间质纤维化的作用及机制研究

Effect of Chebulagic Acid on Renal Interstitial Fibrosis and Its Mechanism

目的探讨诃子酸抗肾间质纤维化的作用及分子机制。方法36只雄性C57/BL小鼠被随机分为假手术组、模型组和诃子酸组。以单侧输尿管结扎术复制肾脏纤维化模型。诃子酸组造模后给予诃子酸灌胃给药。连续给药21天后取血液、肾脏组织。检测肾脏质量指数和血尿素氮(BUN)、血肌酐(Scr)。HE和Masson染色观察肾脏组织病理变化。免疫组化和Western blot检测相关蛋白表达水平。体外实验中,取人肾小管上皮HK-2细胞缺氧处理模拟肾脏纤维化,分为对照组、缺氧组、给药组(缺氧后给予诃子酸)以及转染组(转染siRNA-Klotho以敲减Klotho,然后缺氧并给予诃子酸),Western blot检测相关蛋白表达。结果诃子酸组肾脏质量指数、肾脏组织Klotho蛋白表达水平高于模型组,肾脏组织成纤维细胞生长因子2(FGF2)蛋白表达水平以及血清BUN和Scr水平低于模型组(P<0.05)。诃子酸组较模型组的病理变化明显缓解,纤维化程度以及转化生长因子-β1、α-平滑肌肌动蛋白表达水平降低。给药组的细胞活力、Klotho蛋白表达水平高于缺氧组,乳酸脱氢酶、FGF2蛋白表达水平低于缺氧组(P<0.05)。转染组Klotho蛋白表达水平低于给药组,FGF2蛋白表达水平高于给药组(P<0.05)。结论诃子酸可有效缓解肾间质纤维化水平,其机制可能是通过调控Klotho/FGF2通路发挥作用。诃子酸是潜在的治疗肾脏纤维化药物。

Objective To study the effect and molecular mechanism of Chebulagic acid on renal interstitial fibrosis.Methods A total of 36 male C57/BL mice were randomly divided into sham operation group,model group and Chebulagic acid group.Renal fibrosis model was established by unilateral ureteral ligation.Chebulagic acid group was given gavage administration of Chebulagic acid after modeling.Blood and kidney tissues were collected after 21 d continuous administration.The kidney weight index,serum urea nitrogen(BUN)and creatinine(Scr)were measured.HE and Masson staining were used to observe the renal pathological changes.Immunohistochemistry and Western blot were used to detect the protein expressions.In vitro experiments,human renal tubular epithelial HK-2 cells were treated with hypoxia treated to simulate renal fibrosis,Divided into control group,hypoxic group,administration group(administered with Chebulagic acid after hypoxia)and transfection group(transfected with siRNA-Klotho to knock down Klotho,then hypoxic and administered with Chebulagic acid),Western blot was used to detect the expression of proteins.Results Kidney quality index and Klotho protein expression levels in Chebulagic acid group were higher than those in model group,and renal tissue fibroblast growth factor 2(FGF2)protein expression levels and serum BUN and Scr levels were lower than model group(P<0.05).Compared with model group,the pathological changes in Chebulagic acid group were significantly alleviated,and the degree of fibrosis and the expression levels of transforming growth factor-β1 andα-smooth muscle actin were reduced.The cell viability and Klotho protein expression levels in administered group were higher than those in hypoxic group,and the lactate dehydrogenase and FGF2 protein expression levels were lower than those in hypoxic group(P<0.05).The expression level of Klotho protein in the transfection group was lower than that in administration group,and the expression level of FGF2 protein was higher than that in administration group(P<0.05).Conclusion Chebulagic acid can effectively relieve renal interstitial fibrosis,and its mechanism may be through the regulation of Klotho/FGF2 pathway.Chebulagic acid is a potential treatment for renal fibrosis.