γ干扰素与甲强龙合用对人胚肺成纤维细胞的影响

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

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β1mRNA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-β1mRNA、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-β1mRNA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。

AIM： To look for a better method to deal with interstitial lung disease, interferon - gamma （IFN- γ） combined with methylprednisolane （M- pred） to influence human embryonic lung fibroblast on proliferation, collagen synthesis and the expression of transforming growth factor - β1 （TGF - β1 ） protein and mRNA were investigated. METHODS： Exponentially growing cells were preincubated for 48 h before harvested. The microcuhure tetraznlium （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 - β1 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 - β1 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.