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香烟提取物对人支气管平滑肌细胞DNA的损伤作用及细胞应激的影响 预览 被引量:7

Effects of cigarette smoke extract on DNA damage and cell stress in human bronchi smooth muscle cells
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摘要 目的:探讨香烟提取物(CSE)对人支气管平滑肌细胞DNA损伤和细胞应激(热休克蛋白70表达)的影响.方法: 以1 mL培养液中含30 mL烟雾作为原液,分别以1/16、1/10、1/8、1/6和1/4浓度与正常人支气管平滑肌细胞培养3 h,采用单细胞琼脂糖凝胶电泳(即彗星实验)和Western印迹技术检测细胞DNA损伤和热休克蛋白70(HSP70)的表达.结果: 随着培养液中CSE浓度的增加,细胞受损的百分率(拖尾细胞百分率)及受损的程度(尾长)逐渐增加,除1/16原液外,其它与阴性对照之间均有显著差异(P<0.05).随着培养液中CSE浓度的改变,结果发现含1/16和1/10原液的CSE刺激后HSP70的表达有轻度增高的趋势,而含1/8原液-1/4原液的CSE刺激后,HSP70的表达逐渐降低.其中含1/6原液组和含1/4原液组与阴性对照组之间HSP70的表达有显著差异(P<0.05).结论: CSE可导致支气管平滑肌细胞DNA的损伤并伴有细胞保护蛋白(HSP70)合成的减少,可能与慢性阻塞性肺疾病(COPD)的发病有关. AIM: To investigate DNA damage and cell stress (heat shock protein 70 expression) in human bronchi smooth muscle cells by cigarette smoke extract (CSE) in vitro. METHODS: 30 mL smog was dissolved in 1 mL culture medium as stock solution of CSE. Human bronchi smooth muscle cells were cultured 3 hours with 1∶16, 1∶10, 1∶8, 1∶6 and 1∶4 of CSE. The DNA damage and HSP70 expression were determined by single cell gel assay (comet assay) and Western blot, respectively. RESULTS: Associated with rising CSE concentration, DNA damage aggravated. Compared with the untreated group, except 1∶16 of CSE, the level of DNA damage was significantly different (P<0.05). The level of HSP70 expression rised in 1∶16 and 1∶10 of CSE, but it gradually decreased in others. There was significantly different in the fifth and sixth group with negative control (P<0.05). CONCLUSION: CSE results in DNA damage and decreasing of HSP70 expression, which may be related with chronic obstructive pulmonary diseases. [
作者 谢俊刚 徐永健 张珍祥 倪望 陈仕新 XIE Jun-gang, XU Yong-jian, ZHANG Zhen-xiang, NI Wang, CHEN Shi-xin(Department of Respiratory Medicine, Tongji Hospital of Tongji Medical College, Huazhong Science and Technology University, Wuhan 430030, China) [
出处 《中国病理生理杂志》 CAS CSCD 北大核心 2004年第11期 2028-2032,共5页 Chinese Journal of Pathophysiology
基金 国家科技攻关项目,卫生部部属(管)医疗机构临床学科重点项目
关键词 烟雾吸入损伤 DNA损伤 热休克蛋白质70 支气管 Smoking inhalation injury DNA damage Heat-shock proteins 70 Bronchi
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