上调PTEN表达对血管紧张素Ⅱ诱导心肌成纤维化细胞增殖迁移的影响

摘要

Objective To investigate the influence of up-regulation of PTEN expression on proliferation and migration of cardiac fibroblasts (CFB) induced by angiotensin Ⅱ (Ang Ⅱ). Methods CFB were divided into blank control group (CFB without treatment), Ang Ⅱ group (CFB treated with Ang II), Ang Ⅱ+NC group (CFB treated with Ang II after transfection of pcDNA3.1) and Ang Ⅱ+PTEN group (CFB treated with Ang Ⅱ after transfection of pcDNA3 and 1-PTEN). The expression of PTEN was detected by using Western blotting assay after CFB transfected with liposome 2000 and treated with Ang Ⅱ. The proliferation of CFB was detected by using MTT, and periodic changes of CFB were measured by using flow cytometer. The ability of migration was detected by using Transwell chamber test, and content of collagen I and collagen Ⅲ were detected by using ELISA. Results The expression of PTEN and G0/G1 phase CFB ratio decreased, and percentages of S phase CFB and G2/M phase CFB, ability of proliferation and migration of CFB and content of collagen I and collagen III increased in Ang Ⅱ group, Ang Ⅱ+NC group and Ang Ⅱ+PTEN group compared with blank control group. The expression of PTEN and G0/G1 phase CFB ratio increased, and S phase CFB and G2/M phase CFB, ability of proliferation and migration of CFB and content of collagen Ⅰ and collagen Ⅲ decreased in Ang Ⅱ+PTEN group compared with Ang Ⅱ group. The difference had no statistical significance between Ang Ⅱ group and Ang Ⅱ+NC group. Conclusion The expression of PTEN decreases in CFB induced by Ang Ⅱ, and up-regulation of PTEN expression can inhibit the proliferation and migration of CFB induced by Ang Ⅱ and synthesis of collagen.%目的 探讨上调PTEN表达对血管紧张素Ⅱ(Ang Ⅱ)诱导心肌成纤维化细胞增殖迁移的影响.方法 将心肌成纤维化细胞随机分为空白对照组(未处理)、Ang Ⅱ组(加Ang Ⅱ处理)、Ang Ⅱ+NC组(转染pcDNA3.1后,给予Ang Ⅱ处理)和Ang Ⅱ+PTEN组(转染pcDNA3,1-PTEN后,给予Ang Ⅱ处理),采用脂质体2000转染细胞,并经Ang Ⅱ处理后,Western blot检测各组细胞中PTEN蛋白的表达, MTT法检测细胞增殖能力,流式细胞仪检测细胞的周期变化,Transwell小室实验检测细胞的迁移能力;ELISA检测胶原蛋白Ⅰ和Ⅲ含量.结果 与空白对照组相比,Ang Ⅱ组、Ang Ⅱ+NC组和Ang Ⅱ+PTEN组细胞中PTEN蛋白表达和G0/G1期细胞比例降低,而S期和G2/M期细胞比例、细胞增殖和迁移能力以及胶原蛋白Ⅰ和Ⅲ含量升高;与Ang Ⅱ组相比,Ang Ⅱ+PTEN组细胞中PTEN蛋白表达和G0/G1期细胞比例升高,而S期和G2/M期细胞比例、细胞增殖和迁移能力以及胶原蛋白Ⅰ和Ⅲ含量均降低;Ang Ⅱ组与AngⅡ+NC组间差异无统计学意义.结论 PTEN在Ang Ⅱ诱导心肌成纤维化细胞中表达下降,上调其表达可抑制Ang Ⅱ诱导心肌成纤维化细胞增殖迁移和胶原蛋白的合成.