活性维生素D3抑制大鼠间质成纤维细胞激活作用的研究

1,25-dihydroxyvitamin D3 suppresses fibronectin production induced by TGF-β1 in renal interstitial fibroblasts

关键词:  活性维生素D₃  成纤维细胞  转化生长因子β1  纤维连接蛋白

Key words:  1,25-(OH)₂VitD₃  fibroblast  transforming growth factor β1  fibronectin

摘要:

目的通过观察活性维生素D3(1,25-(OH)2VitD₃,活性VitD3)抑制转化生长因子β1(Transforming growthfactorβ1,TGF-β1)诱导大鼠肾脏间质成纤维细胞(NRK-49F)合成纤维连接蛋白(Fibronectin,FN)作用的研究,探讨活性VitD3阻止肾组织纤维化作用的机制。方法体外培养的NRK-49F细胞,分为对照组、不同时间TGF-β1处理组、活性VitD3处理组、不同剂量活性VitD₃与TGF-β1联合处理组。通过蛋白免疫印迹、免疫荧光染色等实验方法,分别以α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)作为成纤维细胞被激活指标、FN作为细胞外基质合成的检测指标,比较不同浓度活性VitD₃对经TGF-β1刺激的NRK-49F细胞表达α-SMA以及合成FN的变化。结果TGF-β1可以诱导NRK-49F的α-SMA和FN蛋白表达明显增加,并呈一定的时间依赖性,同时,还可影响α-SMA和FN蛋白结构的组装和沉积;活性VitD₃单独作用对α-SMA和FN蛋白表达和组装无明显影响,而活性VitD₃与TGF-β1同时作用于NRK-49F时,活性VitD₃可以部分拮抗TGF-β1的效应,抑制α-SMA和FN蛋白表达,并呈剂量依赖关系。结论活性VitD3可以部分拮抗TGF-β1诱导的肾成纤维细胞的活化,抑制细胞外基质FN的合成分泌。

Abstract:

Objective To investigate the effect of 1,25-(OH)₂ VitD₃ on the production of fibronectin induced by transforming growth factor β1(TGF-β1) in renal interstitial fibroblasts.Methods To study the effect of 1,25-(OH)₂ VitD3 and TGF-β1 on ECM synthesis.The renal interstitial fibroblast NRK-49F cells were divided into control group,TGF-β1 group,VitD₃ group and TGF-β1 plus VitD₃ group.The de novo expression of α-smooth muscle actin(α-SMA) and fibronectin were detected by western blot analysis and immunofluorescence.Results TGF-β1 induced de novo expression of α-SMA and production of fibronection in a time-dependent manner.TGF-β1 also affected assembly of α-SMA and deposition of fibronectin.1,25-(OH)₂ VitD₃ alone had no effects on α-SMA and fibronectin expression.Whereas,1,25-(OH)₂VitD₃ portially obliterated T expression of α-SMA and fibronectin in a dose-dependent manner.Conclusion The results indicate that 1,25-(OH)₂ VitD₃ can partially inhibit renal interstitial fibroblast activation and fibronectin synthesis induced by TGF-β1.