摘要
目的:探讨转化生长因子β1(TGF-β1)在精氨酸加压素(AVP)诱导的心肌成纤维细胞(CFs)向肌成纤维细胞(MFs)转化中的作用。方法:用胰酶消化法分离培养SD仔鼠的CFs,将CFs分别与不同浓度的AVP、不同浓度的TGF-β1或添加了不同浓度的抗TGF-β1中和抗体的1×10-6mmol/L AVP共同孵育48 h后,用3H-脯氨酸掺入法检测CFs胶原合成的功能,用Western blot检测CFs中平滑肌肌动蛋白α(α-SMA)的表达量,用ELISA法检测CFs中TGF-β1的合成。结果:AVP可浓度依赖性地诱导CFs的3H-脯氨酸掺入率和α-SMA的表达量增加,其中1×10-6mmol/L AVP组CFs的3H-脯氨酸掺入率和α-SMA的表达量显著高于对照组(P<0.05)。AVP也可浓度依赖性地诱导CFs中内源性TGF-β1的合成增加,其中在1×10-6mmol/L AVP刺激下,CFs合成显著增加的内源性TGF-β1刚好达到外源性TGF-β1诱导CFs向MFs转化所需要的剂量(>2 ng/ml)。抗TGF-β1中和抗体虽然能够显著抑制在10-6mmol/L AVP诱导下CFs的3H-脯氨酸掺入率和α-SMA的表达量(P<0.05),但却不能将其抑制到与对照组相近的水平(P<0.05)。结论:AVP刺激下CFs中内源性TGF-β1合成的增加可部分地介导AVP诱导的CFs向MFs转化。
AIM：To investigate the role of transforming growth factor β1（TGF-β1） in the differentiation of cardiac fibroblasts（CFs） into myofibroblasts（MFs） induced by arginine vasopressin（AVP）.METHODS：CFs were isolated by trypsin digestion method and co-incubated for 48 h with different concentrations of AVP,different concentrations of TGF-β1,or 1×10-6 mmol/L AVP supplemented with different concentrations of TGF-β1 neutralizing antibody,respectively.3H proline incorporation was used to measure collagen synthesis,Western blot analysis was used to measure α-smooth muscle actin（α-SMA） expression,and ELISA was used to measure TGF-β1 secretion in CFs.RESULTS：AVP induced the increases of 3H proline incorporation and α-SMA expression dose dependently in CFs,and 1×10-6 mol/L AVP notably increased 3H proline incorporation and α-SMA expression compared with those in control group（P0.05）.AVP induced endogenous TGF-β1 secretion of CFs dose dependently,and the notably increased endogenous TGF-β1 induced by 1×10-6 mol/L AVP was well within the range for exogenous TGF-β1 to cause differentiation of CFs into MFs（2 ng/ml）.Although 1×10-6 mol/L AVP-induced increases of 3H proline incorporation and α-SMA expression could be notably inhibited by TGF-β1 neutralizing antibody（P0.05）,they were still higher than those in control group（P0.05）.CONCLUSION：AVP can induce differentiation of CFs into MFs partly through the increase of endogenous TGF-β1 synthesis.
出处
《心脏杂志》
CAS
2012年第1期7-11,共5页
Chinese Heart Journal
基金
国家自然科学基金项目资助(30800470)
