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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。
Objective: To investigate the role of transforming growth factor β1 (TGF-β1) in the conversion of arginine vasopressin (AVP) -induced myocardial fibroblasts (CFs) into myofibroblasts (MFs). Methods: The CFs of SD rats were isolated and cultured by trypsin digestion. CFs were incubated with different concentrations of AVP, different concentrations of TGF-β1 or with different concentrations of anti-TGF-β1 neutralizing antibody (1 × 10-6mmol / L AVP for 48 h, the function of collagen synthesis in CFs was detected by 3H-proline incorporation, and the level of smooth muscle actin α (α-SMA) expression in CFs was detected by Western blot. Synthesis of TGF-β1. RESULTS: AVP induced 3H-proline incorporation and α-SMA expression in CFs in a concentration-dependent manner. The incorporation of 3H-proline in CFs of 1 × 10-6 mmol / L AVP and α The expression of-SMA was significantly higher than that of the control group (P <0.05). AVP also induced a concentration-dependent increase in the synthesis of endogenous TGF-β1 in CFs, in which the endogenous TGF-β1 significantly increased in the synthesis of CFs at 1 × 10 -6 mmol / L AVP just reached the level of exogenous TGF -β1 The dose (> 2 ng / ml) required to induce the conversion of CFs to MFs. Although the anti-TGF-β1 neutralizing antibody could significantly inhibit 3H-proline incorporation and α-SMA expression of CFs induced by 10-6mmol / L AVP (P <0.05), they could not inhibit Similar to the control group (P <0.05). CONCLUSIONS: The increased synthesis of endogenous TGF-β1 in CFs stimulated by AVP partially mediated the AVP-induced conversion of CFs to MFs.