γ干扰素与甲强龙合用对人胚肺成纤维细胞的影响

目的：观察γ干扰素(IFN-γ)与甲强龙(M-pred)联合应用对人胚肺成纤维细胞(HELF)增殖、胶原合成及转化生长因子β1 mRNA与蛋白表达的影响。方法：噻唑蓝（MTT）法观察HELF的生长抑制率，免疫细胞化学方法检测增殖细胞核抗原（PCNA）表达，碱水解法测定羟脯氨酸含量，半定量RT-PCR法、Western印迹技术检测TGF-β1 mRNA、蛋白变化。结果： M-pred与不同浓度IFN-γ单独或联合应用均具有抑制HELF增殖、降低PCNA、羟脯氨酸、TGF-β1 mRNA及蛋白表达作用， IFN-γ的作用随其浓度增加而增强（P<0.05）。析因分析表明IFN-γ与M-pred联合应用具有协同作用（P<0.05）。结论：IFN-γ与M-pred联合应用治疗肺纤维化时可增强疗效，减少各自用药量，可能成为治疗肺纤维化的理想方法。

Influence of IFN-γ combined with M-pred On human embryonic lung fibroblasts

AIM: To look for a better method to deal with interstitial lung disease, interferon-gamma (IFN-γ) combined with methylprednisolone (M-pred) to influence human embryonic lung fibroblast on proliferation, collagen synthesis and the expression of transforming growth factor-β1 (TGF-β1) protein and mRNA were investigated. METHODS: Exponentially growing cells were preincubated for 48 h before harvested. The microculture tetrazolium (MTT) assay was used to measure the inhibition ratios of M-pred combined with different concentrations of IFN-γ. The expression of proliferation cell nuclear antigen (PCNA) was detected by immunocytochemical analysis. Hydroxyproline kit was adopted to detect collagen synthesis. The expressions of TGF-β1 mRNA and protein were detected respectively by RT-PCR and Western blotting. RESULTS: Methylprednisolone, IFN-γ as well as the combination of methylprednisolone and IFN-γ inhibited the proliferation of HELF and the expression of PCNA in comparison with control group (P<0.05). That also decreased the expression of hydroxyproline, inhibited the expression of TGF-β1 mRNA and protein (P<0.05). The effectiveness of IFN-γ became stronger when the concentration increased. The synergistic effect between IFN-γ and methylprednisolone was observed (P<0.05). CONCLUSION: The combination effect of IFN-γ and methylprednisolone is augmented compared with using IFN-γ or methylprednisolone alone, suggesting that the combination use of both drugs has better anti-fibrous degeneration effect than use either alone.