γ-干扰素对人气道壁血管平滑肌细胞ADAM33基因表达的影响

Effect of γ-interferon on the expression of ADAM33 gene in human airway wall vascular smooth muscle cells

目的探讨以不同浓度及不同时间γ-干扰素(IFN-γ)刺激人气道血管壁平滑肌细胞对去整合素金属蛋白酶33(ADAM33)基因表达的影响。方法选择2013年1月-2015年5月在新疆医科大学第一附属医院胸外科行手术治疗的肺大泡患者的患肺切除的肺组织,先以4个不同浓度的IFN-γ((0.1、1、10和100 ng/mL)刺激培养的人气道壁血管平滑肌细胞,然后利用实时定量反转录多聚酶链反应(RT-PCR)法测定ADAM33 mRNA的表达变化,以及利用蛋白印迹法(Western Blot)来测定ADAM33蛋白表达的变化。另用恒定浓度(100 ng/mL)的IFN-γ刺激培养的人气道壁血管平滑肌细胞,观察不同时间(0、6、12、24、48和72 h)ADAM33 mRNA和蛋白表达的变化。结果在人气道壁血管平滑肌细胞中可以检测到ADAM33 mRNA的表达,ADAM33 mRNA的表达量随着刺激浓度的增加有逐渐下降的趋势。与对照组相比,不同浓度IFN-γ刺激下ADAM33蛋白表达量分别为(1.201±0.217)、(0.696±0.109)、(0.522±0.087)、(0.348±0.043)和(0.261±0.065),呈下降趋势,差异有统计学意义(P<0.05),ADAM33蛋白条带灰度与对照组相比逐渐下降,差异有统计学意义。使用一定浓度(100 ng/mL)IFN-γ刺激体外培养的人气道壁血管平滑肌细胞时,ADAM33 mRNA的表达量随着刺激时间的推移,先呈逐渐下降的趋势后转为逐渐上升的趋势,各组的ADAM33 mRNA表达无统计学差异(P>0.05)。随着不同刺激时间的变化,ADAM33蛋白表达量变化趋势无明显规律性,无统计学差异(P>0.05)。结论 IFN-γ能影响人气道壁血管平滑肌细胞ADAM33基因的表达。

Objective To investigate the effect of interferon-gamma(IFN-γ) irritation on the expression of disintegrin metalloproteinase 33(ADAM33) gene in human airway vascular smooth muscle cells at different concentrations and at different times. Methods Lung tissues from patients with alveoli who underwent lung resection in the First Affiliated Hospital of Xinjiang Medical University from January 2013 to May 2015 were irritated with four different concentrations of IFN-γ(0.1, 1, 10 and 100 ng/mL). The expression of ADAM33 was determined by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR). The changes of ADAM33 protein expression were detected by Western Blotting. In addition, cultured human airway wall vascular smooth muscle cells were irritated with IFN-γ at a constant concentration(100 ng/mL). The changes of ADAM33 gene and protein expression were observed at different time points(0, 6, 12, 24, 48 and 72 h). Results The expression of ADAM33 was detected in human airway wall vascular smooth muscle cells, and the expression of ADAM33 decreased with the increase of irritation concentration of IFN-γ. Compared with the control group, the expression levels of ADAM33 protein in different concentrations of IFN-γ irritation were(1.201±0.217),(0.696±0.109),(0.522±0.087),(0.348±0.043) and(0.261±0.065). The difference was statistically significant(P<0.05). The gray level of ADAM33 protein band decreased gradually compared with the control group, and the difference was statistically significant. When cultured human airway wall vascular smooth muscle cells were irritated by IFN-γ at 100 ng/mL, the expression of ADAM33 mRNA decreased gradually and then increased gradually with the passage of irritation time. There was no significant difference in the expression of ADAM33 mRNA among groups(P>0.05). With the change of irritation time, the expression of ADAM33 protein had no obvious regularity and no statistical difference(P>0.05). Conclusion IFN-γ can affect the expression of ADAM33 gene in human airway wall vascular smooth muscle cells.