目的观察香烟烟雾长期染毒大鼠肺组织、气管上皮细胞中晚期糖基化终产物受体(RAGE)和钙结合蛋白(S100A6)的表达。方法健康雄性Wistar大鼠18只,随机分为对照组、低(烟雾浓度20%)和高(烟雾浓度60%)浓度组,每组6只。动式气体染毒装置每天染毒2次,每次30min,一周5d,共染毒43周。分离大鼠气管和肺组织,分离原代上皮细胞,收集并提取总蛋白;Western blot方法检测RAGE和S100A6蛋白的表达。同时测定正常的人支气管上皮细胞株BEAS-2B和肺癌细胞株A549中2种蛋白的表达情况。结果气管组织贴块培养法分离出的上皮细胞纯度为(93.56±1.48)%;烟雾染毒组大鼠肺组织及气管上皮细胞中RAGE和S100A6蛋白的表达均显著增高。同时,肺癌细胞A549中RAGE及S100A6蛋白的表达量明显高于正常人支气管上皮细胞BEAS-2B。结论RAGE和S100A6有望作为香烟烟雾暴露致肺损伤的生物学标志。
Objective To determine the change of RAGE and $100A6 in lung tissue and tracheal epithelial cells of rats long-term exposed to cigarette smoke. Methods Wistar rats were randomly divided into 3 groups. The low and high concentration groups were exposed to 20 % and 60 % of cigarette smoke in chamber gas respectively, with the control group in a non-sraoke environment. The exposure was conducted 30 minutes each time and twice a day, 5 days a week for 43 weeks in total. Cultures were obtained from a type I rat collagen substratum and the expression of RAGE and S100A6 were analyzed by using western blotting. Results The purity of the epithelium was (93.56 ± 1.48) %. The expressions of RAGE and S100A6 in samples from the exposure groups increased significantly compared to the control group, especially in the high concentration group. Conclusions RAGE and S100A6 could be used as potential exposure biomarkers for cigarette smoke.