蛋白酪氨酸磷酸酶SHP-2对Ang Ⅱ刺激的心肌成纤维细胞增殖的影响

摘要

目的:探讨含有Src同源结构域2的蛋白酪氨酸磷酸酶2 (Src homology 2 domain-containing protein tyrosine phosphatase 2,SHP-2) 对血管紧张素Ⅱ (angiotensin Ⅱ,Ang Ⅱ) 刺激的心肌成纤维细胞(cardiac fibroblasts,CFs)增殖的作用.方法:差速贴壁法体外培养心肌成纤维细胞,以波形蛋白(vimentin)鉴定CFs纯度;MTT法检测Ang Ⅱ作用下心肌成纤维细胞增殖率,采用重组腺病毒过表达SHP-2和SHP-2抑制剂NSC-87877分别对Ang Ⅱ作用下的细胞增殖的影响.结果:Ang Ⅱ对CFs增殖的促进作用有剂量依赖性,其促进细胞增殖的最高浓度为10-7 mol/L;在AngⅡ的刺激下,SHP-2可以促进心肌成纤维细胞增殖,并且突变体组比野生型组增殖更明显(P<0.01).SHP-2抑制剂NSC-87877达到50 μmol/L时可以明显抑制Ang Ⅱ刺激下的CFs增殖.结论:Ang Ⅱ的促CFs增殖作用是通过SHP-2调控的.%AIM: To investigate the potential role of Src homology 2 domain - containing protein tyrosine phos-phatase 2 ( SHP - 2 ) in the proliferation of cardiac fibroblasts induced by angiotensin II ( Ang II ). METHODS: The neonatal rat cardiac fibroblasts were separated by trypsin digestion. The cardiac fibroblasts were identified by vimentin of immunochemical staining. The proliferation of the cardiac fibroblasts with or without Ang II stimulation was measured by MTT assay. Adenoviruses containing SHP -2 gene were transfected into cardiac fibroblasts to overexpress SHP -2. SHP - 2 was inhibited by its inhibitor NSC - 87877. RESULTS: The proliferation of the cardiac fibroblasts was increased in a dose - dependent manner by the stimulation of Ang II and the maximum concentration of Ang II for cell proliferation was 10 mol/L. SHP - 2 promoted the proliferation of cardiac fibroblasts under the stimulation of Ang II. The proliferation rate in mutant group was higher than that in wild - type group ( P <0. 01 ). Inhibition of SHP - 2 by NSC - 87877 attenuated the proliferation. CONCLUSION: The growth promoting effect of Ang II on cardiac fibroblasts is regulated by SHP-2.