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目的探讨c-Jun氨基末端激酶(JNK)特异性抑制剂SP600125对血管紧张素Ⅱ(AngⅡ)诱导的肾小球系膜细胞(MC)转化生长因子-β1(TGF-β1)和纤维连接蛋白(FN)表达的影响。方法体外分离、培养人MC,应用AngⅡ刺激或应用SP600125预处理后再应用AngⅡ刺激。于实验终点抽提细胞总蛋白或收集培养上清,采用Westernblot检测MCJNK、胞外信号调节激酶(ERK1/2)及p38丝裂原活化蛋白激酶(p38MAPK)活性;应用ELISA法检测培养上清中TGF-β1和FN的分泌。结果SP600125可呈剂量依赖性抑制AngⅡ诱导的JNK活化,10μmol/LSP600125对JNK活性的抑制率为75%,而20μmol/LSP600125几乎完全阻断AngⅡ诱导的JNK活化。AngⅡ可促进MCERK1/2和p38活化,而SP600125对AngⅡ诱导的ERK1/2和p38活化无明显影响。AngⅡ显著促进MCTGF-β1和FN分泌,而SP600125则呈时间依赖性和剂量依赖性阻断AngⅡ诱导的MCTGF-β1和FN分泌,20μmol/LSP600125几乎完全抑制了AngⅡ诱导的TGF-β1和FN分泌。结论JNK特异性抑制剂SP600125可阻断AngⅡ诱导的JNK活化及TGF-β1和FN的表达,可能有一定的治疗作用。
Objective To investigate the effect of SP600125, a specific inhibitor of c-Jun N-terminal kinase (JNK), on the expression of transforming growth factor-β1 (TGF-β1) and fibronectin (TGFβ1) induced by angiotensin Ⅱ FN) expression. METHODS: Human MCs were isolated and cultured in vitro. AngⅡ was stimulated by Ang Ⅱ or SP600125 pretreatment. At the end of the experiment, total cellular protein was extracted or culture supernatant was collected. The activity of MCJNK, ERK1 / 2 and p38 mitogen-activated protein kinase (p38MAPK) were detected by Western blot. Secretion of TGF-β1 and FN. Results SP600125 inhibited Ang Ⅱ-induced JNK activation in a dose-dependent manner. The inhibition rate of JNK was 75% at 10μmol / LSP600125 and almost completely blocked AngⅡ-induced JNK activation at 20μmol / LSP600125. AngⅡ promoted the activation of MCERK1 / 2 and p38, while SP600125 had no significant effect on AngⅡ-induced ERK1 / 2 and p38 activation. AngⅡ significantly promoted the secretion of MCTGF-β1 and FN, while SP600125 blocked AngⅡ-induced MCTGF-β1 and FN secretion in a time-and dose-dependent manner. 20μmol / LSP600125 almost completely inhibited AngⅡ-induced TGF-β1 and FN secretion. Conclusion SP600125, a specific inhibitor of JNK, can block the activation of JNK and the expression of TGF-β1 and FN induced by AngⅡ, which may have a therapeutic effect.