TAK1通过调控p38 MAPK信号通路影响肾小管上皮细胞纤维化  

TAK1 regulates fibrosis of renal tubular epithelial cells by regulating p38 MAPK signaling pathway

作　　者：丁文飞 宋磊[2] 刘海芸[3] 秦建华 曹灵 高利超 欧三桃 吴蔚桦 DING Wen-fei;SONG Lei;LIU Hai-yun;QIN Jian-hua;CAO Ling;GAO Li-chao;OU San-tao;WU Wei-hua(Department of Nephrology, Affiliated Hospital of Southwest Medical University, Luzhou 646000 , China;Department of Nephropathy, Jiangxi Provincial People’s Hospital, Nanchang 330006 , China;Department of Laboratory Medicine, Jiangxi Provincial People’s Hospital, Nanchang 330006 , China)

机构地区：[1]1西南医科大学附属医院肾病内科,四川泸州646000 [2]江西省人民医院肾病内科,江西南昌330006 [3]江西省人民医院检验科,江西南昌330006

出　　处：《中国病理生理杂志》2019年第7期1248-1253,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81200533);西南医科大学科研基金资助项目(No.2017-ZRQN-021);西南医科大学附属医院科研基金资助项目(No.2017-PT-33)

摘　　要：目的:探讨转化生长因子β激活激酶1(TAK1)对肾小管上皮细胞纤维化的影响及机制。方法:以肾小管上皮细胞HK-2作为研究对象,用转化生长因子β1(TGF-β1)诱导肾小管上皮细胞纤维化,以real-time PCR和Western blot检测细胞中TAK1表达的变化。用TAK1 shRNA慢病毒感染肾小管上皮细胞,以real-time PCR和Western blot检测其对TGF-β1刺激下肾小管上皮细胞中TAK1表达的影响以检测干扰效果。ELISA法检测细胞分泌的I型胶原和Ⅲ型胶原水平,Western blot法检测细胞中α-平滑肌肌动蛋白(α-SMA)、结缔组织生长因子(CTGF)和磷酸化的p38 MAPK(p-p38 MAPKThr 180/Tyr 182)的蛋白水平。用p38 MAPK激活剂处理已敲减TAK1表达的肾小管上皮细胞,检测其对细胞分泌I型胶原和Ⅲ型胶原的影响及对细胞中α-SMA、CTGF和p-p38 MAPKThr 180/Tyr 182蛋白水平的影响。结果:TGF-β1可以明显上调肾小管上皮细胞中TAK1的表达水平。TAK1 shRNA可明显下调TGF-β1刺激下的肾小管上皮细胞中TAK1的表达水平。TGF-β1处理后的肾小管上皮细胞分泌I型胶原和Ⅲ型胶原增多,细胞中α-SMA、CTGF和p-p38 MAPKThr 180/Tyr 182蛋白水平升高。敲减TAK1表达可以明显抑制TGF-β1诱导的肾小管上皮细胞分泌I型胶原和Ⅲ型胶原,减少细胞中α-SMA、CTGF和p-p38 MAPKThr 180/Tyr 182的蛋白水平(P<0.05)。p38 MAPK激活剂处理可以逆转敲减TAK1表达对肾小管上皮细胞分泌Ⅰ型胶原和Ⅲ型胶原及α-SMA、CTGF和p-p38 MAPKThr 180/Tyr 182蛋白水平的抑制作用。结论:敲减TAK1表达能够通过抑制p38 MAPK信号通路而降低TGF-β1诱导的肾小管上皮细胞纤维化水平。AIM: To investigate the effect of transforming growth factor-β(TGF-β) activated kinase 1(TAK1) on renal tubular epithelial fibrosis. METHODS: The renal tubular epithelial cell line HK-2 was used as the research object. After induced by TGF-β1, real-time PCR and Western blot were used to detect the expression of TAK1 in the HK-2 cells. TAK1 shRNA lentivirus was used to infect HK-2 cells, real-time PCR and Western blot were used to determine the interference effect on TAK1 expression in the HK-2 cells with TGF-β1 stimulation. Under the condition of treating with p38 MAPK activator anisomycin, the levels of type I collagen and type III collagen in the supernatant, and the protein levels of α-smooth muscle actin(α-SMA), connective tissue growth factor(CTGF) and p-p38 MAPKThr 180/Tyr 182 in the HK-2 cells with TAK1 knock-down were determined by ELISA and Western blot, respectively. RESULTS:TGF-β1 significantly increased the expression of TAK1 in the HK-2 cells(P<0.05). TAK1 shRNA significantly decreased the expression of TAK1 in the HK-2 cells with TGF-β1 stimulation. Type I collagen and type III collagen secreted by the HK-2 cells after treatment with TGF-β1 were increased, the protein levels of α-SMA, CTGF and p-p38 MAPKThr 180/Tyr 182 were also increased(P<0.05). Knock-down of TAK1 expression significantly inhibited the secretion of type I and type III collagen, reduced the protein levels of α-SMA, CTGF and p-p38 MAPKThr 180/Tyr 182 in the TGF-β1-induced HK-2 cells(P<0.05). Treatment with p38 MAPK activator reversed the inhibitory effect of TAK1 knock-down on the secretion of type I and type III collagens, and the protein levels of α-SMA, CTGF and p-p38 MAPKThr 180/Tyr 182 in the HK-2 cells(P<0.05). CONCLUSION: Knock-down of TAK1 expression attenuates the TGF-β1 induced fibrosis of renal tubular epithelial cells by inhibiting p38 MAPK signaling pathway.

关 键 词：肾小管上皮细胞 转化生长因子β激活激酶1 P38 MAPK信号通路 纤维化 

分 类 号：R692[医药卫生—泌尿科学] R363[医药卫生—外科学]