不同药物对香烟诱导的大鼠肺泡巨噬细胞氧化应激相关因子表达的影响

Effects of different drugs on oxidant stress factors in expression of cigarette induced alveolar macrophage

目的研究地塞米松、氨茶碱、盐酸氨溴索和红霉素对香烟诱导的大鼠肺泡巨噬细胞γ-谷氨酰半胱氨酸合成酶(γ-GCS)、转化生长因子(TGF)-β1和活化蛋白(AP)-1亚单位c-fos mRNA及其蛋白表达的影响,探讨这几种药物在慢性阻塞性肺疾病抗氧化治疗中的作用及其机制。方法通过支气管肺泡灌洗获取大鼠肺泡巨噬细胞,随机分为6组:对照组、香烟组、地塞米松组、氨茶碱组、氨溴索组和红霉素组。后4组分别加入地塞米松、氨茶碱、盐酸氨溴索和红霉素,2h后除对照组外,每组均加入香烟烟雾提取物,对照组加入磷酸盐缓冲液。于不同时间点收取细胞爬片,分别采用原位杂交和免疫细胞化学方法检测肺泡巨噬细胞中γ-GCSh(重链亚单位)、TGF-β1及c-fos mRNA及其蛋白的表达水平。结果与对照组比较,香烟组γ-GCSh、TGF-β1和c-fos mRNA及其蛋白的表达水平明显增高,差异有显著性,P均<0.01。与香烟组比较,地塞米松组和红霉素组γ-GCSh、TGF-β1和c-fos mRNA及其蛋白的表达水平明显降低,差异有显著性,P均<0.01。与香烟组比较,还不能认为氨茶碱组和氨溴索组γ-GCSh、TGF-β1和c-fos mRNA及其蛋白的表达水平差别有统计学意义,P均>0.05。结论香烟烟雾提取物可使大鼠肺泡巨噬细胞γ-GCSh、TGF-β1及c-fos mRNA及其蛋白表达增加,地塞米松和红霉素可使香烟诱导的大鼠肺泡巨噬细胞上述因子表达降低。提示这两种药物可能通过影响这些氧化应激相关因子的表达而发挥一定的抗氧化作用。

[Objective] To evaluate the role of dexamethasone （DEX）, theophylline, ambroxol and erythromycin in the course of treatment for oxidant/antioxidant imbalance in COPD by observing the effects of the four drugs on the expression of γ-GCS, TGF-131 and c-fos mRNA and its proteins in cigarreate smoke extract pretreated rat alveolar macrophage （AM）. [Methods] Acquired the AM of rat by BAL were randomly divided into six groups： control group, CSE group, DEX group, theophylline group, ambroxol group and erythromycin group. The last four groups were added DEX, theophylline, ambroxol and erythromycin respectivetly, two hours later, except control group, others were added CSE, while control group added PBS. Cells were collected at different time. Immunocytochemistry was used to examine the protein expression of γ-GCSh, TGF-β1 and c-fos, respectivetly. In situ hybridization staining was used to examine the mRNA expression of γ-GCSh, TGF-β1 and c-los, respectivetly. [Result] The expression of protein and mRNA of γ-GCSh, TGF-β1 and c-fos in CSE group increased significantly as compared to control group （P 〈0.01, respectively）. The expression of protein and mRNA of γ-RNA, TGF-β1and c-los in DEX and erythromycin group decreased significantly as compared to CSE group （P 〈0.01, respectively）. The expression of protein and mRNA of γ-GCSh, TGF-β1 and c-los in theophylline and ambroxol group did not decrease significantly as compared to CSE group （P 〉0.05, respectively）. [Conclusion] CSE can increase the expression of γ-GCSh, TGF-β1 and c-fos mRNA and its proteins, DEX and erythromycin can decrease the expression of mRNA and its proteins of these factors. The results implicate that DEX and erythromycin may play roles in the antioxidant treatment of COPD