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目的观察肾上腺髓质素(AM)对转化生长因子β1(TGF-β1)促进肺成纤维细胞1型前胶原蛋白α1(Col1α1)、3型前胶原蛋白α1(Col3α1)基因表达及Smad2/3通路的影响,探讨AM对TGF-β1促肺成纤维细胞胶原合成作用的机制。方法体外原代培养人胚肺成纤维细胞(HFLF),利用反转录PCR(RT-PCR)观察AM及TGF-β1对HFLF的Col1α1、Col3α1 mRNA表达的影响;Western blot法观察AM及TGF-β1对HFLF磷酸化Smad2/3(p-Smad2/3)蛋白表达的影响。结果 TGF-β1可以刺激HFLF的Col1α1、Col3α1 mRNA和p-Smad2/3蛋白表达,AM则可抑制基础条件下TGF-β1促进的Col3α1 mRNA表达,同时也抑制Smad2/3蛋白的表达。结论 AM可能通过Smad2/3信号转导途径,抑制TGF-β1促进的HFLF前胶原蛋白的合成作用。
Objective To investigate the effect of adrenomedullin (AM) on the expression of procollagen type 1 (Col1α1) and type 3 procollagen α1 (Col3α1) and the Smad2 / 3 pathway in lung fibroblasts by transforming growth factor-β1 To explore the mechanism of AM on collagen synthesis of TGF-β1-induced lung fibroblasts. Methods Human embryonic lung fibroblasts (HFLF) were cultured in vitro. The effects of AM and TGF-β1 on the expression of Col1α1 and Col3α1 mRNA in HFLF were observed by reverse transcription-polymerase chain reaction (RT-PCR) β1 on HFLF phosphorylation of Smad2 / 3 (p-Smad2 / 3) protein expression. Results TGF-β1 could stimulate the expression of Col1α1, Col3α1 mRNA and p-Smad2 / 3 in HFLF. AM inhibited the expression of Col3α1 mRNA induced by TGF-β1 and the expression of Smad2 / 3 in HFLF. Conclusion AM may inhibit the synthesis of pre-cancerous HFLF collagen by TGF-β1 through the Smad2 / 3 signal transduction pathway.