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
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摘要
目的探讨上调PTEN表达对血管紧张素Ⅱ(AngⅡ)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)、AngⅡ组(加AngⅡ处理)、AngⅡ+NC组(转染pcDNA3.1后,给予AngⅡ处理)和AngⅡ+PTEN组(转染pcDNA3,1-PTEN后,给予AngⅡ处理),采用脂质体2000转染细胞,并经AngⅡ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,AngⅡ组、AngⅡ+NC组和AngⅡ+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AngⅡ组相比,AngⅡ+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;AngⅡ组与AngⅡ+NC组间差异无统计学意义。结论 PTEN在AngⅡ诱导心肌成纤维化细胞中表达下降,上调其表达可抑制AngⅡ诱导心肌成纤维化细胞增殖迁移和胶原蛋白的合成。
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.
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.
引文
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[13] Li NS,Wang ZM,Lin JH. Upregulated expression of PTEN after splenectomy may prevent the progression of liver fibrosis in rats[J].Journal of hepato-biliary-pancreatic sciences,2016,23(1):50-6.
[14]Gao Y,Chu M,Hong J,et al. Hypoxia induces cardiac fibroblast proliferation and phenotypic switch:a role for caveolae and caveolin-1/PTEN mediated pathway[J]. Journal of Thoracic Disease,2014,6(10):1458-68.
[15] Chen D,Liu J,Rui B,et al. GSTpi protects against angiotensin IIinduced proliferation and migration of vascular smooth muscle cells by preventing signal transducer and activator of transcription 3activation[J]. Biochimica et Biophysica Acta(BBA)-Molecular Cell Research,2014,1843(2):454-63
[16] Wang S,Tang L,Zhou Q,et al. miR-185/P2Y6 Axis Inhibits Angiotensin II-Induced Human Aortic Vascular Smooth Muscle Cell Proliferation[J]. Dna&Cell Biology,2017,36(5):377-85.
[17] Li H,Peng X,Wang Y,et al. Atg5-mediated autophagy deficiency in proximal tubules promotes cell cycle G2/M arrest and renal fibrosis[J]. Autophagy,2016,12(9):1472-86.
[18] Xie S,Lu Z,Lin Y,et al. Upregulation of PTEN suppresses invasion in Tca8113 tongue cancer cells through repression of epithelialmesenchymal transition(EMT)[J]. Tumour Biol,2016, 37(5):6681-89.
[19]ZOU G,LIU Y,ZHANG H. Effect of autophagy on AngⅡ-induced proliferation and migration of cardiac fibroblasts[J]. Journal of Guangdong Pharmaceutical University,2015,31(3):32-6.
[2]Zhu Q,Chen D. Advances in relationship between PTEN and myocardial fibrosis[J]. Chinese journal of pathology,2017,46(12):880-3.
[3]赵明,陈锦铷,韩秀敏,等.姜黄素激活自噬对抗ANGⅡ诱导的心肌纤维化发生[J].中国心血管病研究,2017,15(5):470-4.
[4]He Z,Deng Y,Li W,et al. Overexpression of PTEN suppresses lipopolysaccharide-inducedlungfibroblastproliferation,differentiation and collagen secretion through inhibition of the PI3-K-Akt-GSK3beta pathway[J]. Cell&bioscience,2014,4(1):2-13.
[5]McClelland AD,Herman-Edelstein M,Komers R,et al. miR-21promotes renal fibrosis in diabetic nephropathy by targeting PTEN and SMAD7[J]. Clinical science,2015,129(12):1237-49.
[6]Li NS,Wang ZM,Lin JH. Upregulated expression of PTEN after splenectomy may prevent the progression of liver fibrosis in rats[J].Journal of hepato-biliary-pancreatic sciences,2016,23(1):50-6.
[7]赵娜,李娜,刘小军,等. miR-148b参与心肌梗死后心脏纤维化的作用及机制[J].心脏杂志,2017(2):149-53.
[8]Sun Y,Tian H,Wang L. Effects of PTEN on the proliferation and apoptosis of colorectal cancer cells via the phosphoinositol-3-kinase/Akt pathway.[J]. Oncology Reports,2015,33(4):1828-36.
[9]Mense SM,Barrows D,Hodakoski C,et al. PTEN inhibits PREX2-catalyzed activation of RAC1 to restrain tumor cell invasion[J].Science signaling,2015,8(370):ra32.
[10]Guo X,Deng L,Deng K,et al. Pseudogene PTENP1 suppresses gastric cancer progression by modulating PTEN[J]. Anti-Cancer Agents in Medicinal Chemistry(Formerly Current Medicinal Chemistry-AntiCancer Agents),2016,16(4):456-64.
[11]Parapuram SK,Shiwen X,Elliott C,et al. Loss of PTEN expression by dermal fibroblasts causes skin fibrosis[J]. Journal of Investigative Dermatology,2011,131(10):1996-2003.
[12]Zhou J,Zhong J,Lin S,et al. Inhibition of PTEN Activity Aggravates Post Renal Fibrosis in Mice with Ischemia Reperfusion-Induced Acute Kidney Injury[J]. Cellular Physiology&Biochemistry,2017,43(5):1841-54.
[13] Li NS,Wang ZM,Lin JH. Upregulated expression of PTEN after splenectomy may prevent the progression of liver fibrosis in rats[J].Journal of hepato-biliary-pancreatic sciences,2016,23(1):50-6.
[14]Gao Y,Chu M,Hong J,et al. Hypoxia induces cardiac fibroblast proliferation and phenotypic switch:a role for caveolae and caveolin-1/PTEN mediated pathway[J]. Journal of Thoracic Disease,2014,6(10):1458-68.
[15] Chen D,Liu J,Rui B,et al. GSTpi protects against angiotensin IIinduced proliferation and migration of vascular smooth muscle cells by preventing signal transducer and activator of transcription 3activation[J]. Biochimica et Biophysica Acta(BBA)-Molecular Cell Research,2014,1843(2):454-63
[16] Wang S,Tang L,Zhou Q,et al. miR-185/P2Y6 Axis Inhibits Angiotensin II-Induced Human Aortic Vascular Smooth Muscle Cell Proliferation[J]. Dna&Cell Biology,2017,36(5):377-85.
[17] Li H,Peng X,Wang Y,et al. Atg5-mediated autophagy deficiency in proximal tubules promotes cell cycle G2/M arrest and renal fibrosis[J]. Autophagy,2016,12(9):1472-86.
[18] Xie S,Lu Z,Lin Y,et al. Upregulation of PTEN suppresses invasion in Tca8113 tongue cancer cells through repression of epithelialmesenchymal transition(EMT)[J]. Tumour Biol,2016, 37(5):6681-89.
[19]ZOU G,LIU Y,ZHANG H. Effect of autophagy on AngⅡ-induced proliferation and migration of cardiac fibroblasts[J]. Journal of Guangdong Pharmaceutical University,2015,31(3):32-6.