激活TGR5缓解肾缺血再灌注后肾脏纤维化损伤  

Activation of TGR5 Attenuates Renal Fibrosis after Renal Ischemia Reperfusion Injury

作　　者：李蒙 隆罗莎 梁柏恩 徐龙 赵晓朵 王蔚东[2] 李春凌[1] LI Meng;LONG Luo-sha;LIANG Bai-en;XU Long;ZHAO Xiao-duo;WANG Wei-dong;LI Chun-ling(Department of Physiology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou,510080,China;Department of Pathophysiology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou,510080,China)

机构地区：[1]中山大学中山医学院生理教研室,广东广州510080 [2]中山大学中山医学院病理生理教研室,广东广州510080

出　　处：《中山大学学报（医学科学版）》2023年第4期617-624,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81870465,82270744);广东省自然科学基金(2021A1515011482)。

摘　　要：【目的】探讨激活胆汁酸受体TGR5在单肾缺血再灌注损伤合并对侧肾摘除(uIRIx)模型诱导的肾脏纤维化中的作用。【方法】体内实验:将C57BL/6J小鼠随机分成假手术(Sham)组、uIRIx组及uIRIx+石胆酸(LCA)组,每组6只,使用uIRIx模型诱导肾脏纤维化,通过血和尿生化指标评估肾脏功能,利用HE染色评估肾脏损伤程度,使用Masson染色及免疫组化对肾脏纤维化程度进行评估,并用Western Blotting检测肾脏皮质纤维化相关指标蛋白表达;分别在TGR5+/+小鼠及TGR5-/-小鼠中设置Sham组及uIRIx组,每组6只,利用Western Blotting检测各组肾脏纤维化程度。体外实验:在人源肾上皮细胞系HK2细胞中给予TGF-β1诱导促纤维化反应,给予LCA进行药物干预,利用鬼笔环肽染色标记细胞骨架,使用Western Blotting检测HK2细胞内纤维化相关指标蛋白表达。【结果】体内实验:与Sham组相比,uIRIx组小鼠血浆肌酐水平(P=0.007)及尿白蛋白/肌酐比(P=0.041)明显增加,肾脏皮质蛋白TGR5表达(P=0.002)下降,Fibronectin表达(P=0.020)及COL1A1表达(P<0.001)上升,同时伴有肾脏结构受损及胶原沉积加重,LCA干预有效改善肾脏功能,缓解肾脏损伤及纤维化程度;TGR5+/+小鼠与TGR5-/-小鼠相比,uIRIx诱导引起的Fibronectin表达(P<0.001)及COL1A1表达(P=0.001)增加。体外实验:TGF-β1诱导HK2细胞形态改变,细胞骨架解聚重组,促纤维化指标蛋白上调;LCA可有效抑制TGF-β1诱导的细胞形态改变及骨架解聚重组,呈浓度依赖性下调纤维化相关指标蛋白表达。【结论】LCA缓解uIRIx模型诱导的肾脏纤维化,TGR5基因敲除加重uIRIx诱导的肾脏纤维化;在HK2细胞中,LCA缓解TGF-β1诱导的细胞促纤维化反应。研究结果提示激活TGR5缓解缺血再灌注后肾脏纤维化损伤进程。【Objective】To investigate the role of bile acid receptor TGR5 activation in renal fibrosis induced by unilat⁃eral ischemia reperfusion injury and contralateral nephrectomy(uIRIx)model.【Methods】In vivo:C57BL/6J mice were randomly divided into Sham group,uIRIx group and uIRIx+lithcholic acid(LCA)group with 6 mice in each group.Kid⁃ney fibrosis was induced by uIRIx model,kidney function was evaluated by blood and urine biochemical indexes,and the degree of kidney injury was evaluated by HE staining.Masson staining and immunohistochemistry were used to evaluate the degree of renal fibrosis,and Western Blotting was used to detect the expression of related index proteins of renal cortical fibrosis.Sham group and uIRIx group were set in TGR5+/+mice and TGR5-/-mice respectively,with 6 mice in each group.The degree of renal fibrosis in each group was detected by Western Blotting.In vitro:TGF-β1 was administered to induce pro-fibrosis response in human renal tubular epithelial cell line(HK2 cells),LCA was used for drug intervention,cytoskeleton was labeled with phalloidin-FITC staining and the expression of fibrosis related indicator protein in HK2 cells was detected by Western Blotting.【Results】In vivo:Compared with the Sham group,plasma creatinine level(P=0.007)and urinary albumin/creatinine ratio(P=0.041)in uIRIx group were significantly increased,renal cortical protein TGR5 expression(P=0.002)was decreased,Fibronectin expression(P=0.020)and COL1A1 expression(P<0.001)were in⁃creased.At the same time,the kidney structure was damaged and collagen deposition was aggravated.LCA intervention ef⁃fectively improved the kidney function and alleviated the degree of kidney injury and fibrosis.TGR5 gene knockout in⁃creased uIRIx-induced Fibronectin expression(P<0.001)and COL1A1 expression(P=0.001)compared with TGR5+/+mice.In vitro:TGF-β1 induced morphological changes of HK2 cells,cytoskeletal depolymerization and recombination,and promoted the up-regulation of fibrosis index protein.LCA effectively inhibite

关 键 词：Takeda G蛋白耦联受体5 单肾缺血再灌注损伤合并对侧肾摘除 肾脏纤维化 石胆酸 

分 类 号：R692[医药卫生—泌尿科学]