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目的探讨肾上腺髓质素(adrenomedullin,AM)对转化生长因子β1(transforming growth factor-β1,TGF-β1)促肺成纤维细胞增殖及c-myc基因表达的影响。方法体外原代培养人胚肺成纤维细胞(human fetal lung fibroblasts,HFLFs),分为对照组、AM组、TGF-β1组和AM+TGF-β1组,采用MTT法观察各组光吸度(optical density,OD)值;各组提取细胞总RNA,采用反转录PCR观察c-myc基因mRNA表达情况。结果 AM+TGF-β1组HFLFs细胞培养24、48、72h时OD值(0.615±0.054、0.483±0.017、0.675±0.014)明显低于TGF-β1组(0.830±0.012、0.753±0.089、0.929±0.039)(P<0.05),TGF-β1组HFLFs细胞培养72h时OD值(0.929±0.039)明显高于对照组(0.605±0.274)(P<0.05),AM组HFLFs细胞培养24、48h时OD值(0.559±0.019、0.412±0.096)明显低于对照组(0.785±0.081、0.646±0.038)(P<0.05);AM组c-myc mRNA相对表达量(0.450±0.025)明显低于对照组(P<0.01);TGF-β1组c-myc mRNA相对表达量(1.160±0.100)高于对照组(P<0.05);AM+TGF-β1组c-myc mRNA相对表达量(0.340±0.030)低于TGF-β1组(P<0.01)。结论 AM能抑制肺成纤维细胞增殖和c-myc基因表达,AM可能具有抗纤维化作用。
Objective To investigate the effects of adrenomedullin (AM) on proliferation and c-myc gene expression in lung fibroblasts induced by transforming growth factor-β1 (TGF-β1). Methods Primary human fetal lung fibroblasts (HFLFs) were cultured in vitro and divided into control group, AM group, TGF-β1 group and AM + TGF-β1 group. MTT was used to observe the optical absorbance optical density, OD). Total RNA was extracted from each group. The expression of c-myc mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Results The OD values of HFLFs cultured in AM + TGF-β1 group at 24, 48 and 72 h were significantly lower than those in TGF-β1 group (0.830 ± 0.012, 0.753 ± 0.089, 0.929 ± 0.039, ) (P <0.05). The OD value of HFLFs cultured in TGF-β1 group for 72h (0.929 ± 0.039) was significantly higher than that in control group (0.605 ± 0.274, P <0.05) (0.559 ± 0.019,0.412 ± 0.096) was significantly lower than that of the control group (0.785 ± 0.081,0.646 ± 0.038) (P <0.05). The relative expression of c-myc mRNA in AM group (0.450 ± 0.025) was significantly lower than that in the control group <0.01). The relative expression of c-myc mRNA in TGF-β1 group (1.160 ± 0.100) was higher than that in control group (P <0.05) TGF-β1 group (P <0.01). Conclusion AM can inhibit lung fibroblast proliferation and c-myc gene expression, AM may have anti-fibrosis.