N-乙酰半胱氨酸减轻香烟烟雾提取物所致的A549细胞损伤

基础医学与临床 ›› 2014, Vol. 34 ›› Issue (1): 72-77.

N-乙酰半胱氨酸减轻香烟烟雾提取物所致的A549细胞损伤

张强弩¹,焦宗宪²,常嘉琛¹

1. 兰州大学基础医学病理学研究所
2. 兰州大学

收稿日期:2013-01-21 修回日期:2013-05-20 出版日期:2014-01-05 发布日期:2013-12-26
通讯作者: 焦宗宪 E-mail:jiaozongx@lzu.edu.cn
基金资助:
教育部留学回国人员科研启动基金;兰州大学中央高校基本科研业务费专项资金

NAC relieves the A549 cells injury induced by cigarette smoke extract

Received:2013-01-21 Revised:2013-05-20 Online:2014-01-05 Published:2013-12-26
Contact: ZONGXIAN JIAO E-mail:jiaozongx@lzu.edu.cn
Supported by:
The Project Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry.

摘要/Abstract

摘要： 目的研究N-乙酰半胱氨酸（NAC）对香烟烟雾提取物（CSE）暴露后的A549细胞的影响。方法 MTT法测定不同处理后A549细胞的存活率；用PI单染和AnnexinV-FITC/PI双染检测细胞的周期变化和凋亡变化；用荧光探针H2DCFDA检测细胞内活性氧簇(ROS)变化；用实时荧光定量PCR和Western Blot 技术检测胰岛素样生长因子结合蛋白-3(IGFBP-3)的表达变化。结果 CSE致A549细胞存活的A值为0.55±0.04，显著低于对照组的0.78±0.03（P<0.05），NAC处理使A值显著上升至0.67±0.04（P<0.05），并消除了G1期阻滞，降低细胞凋亡率等。IGFBP-3的表达在CSE刺激后，有所升高，在接受NAC保护后，其表达水平有所降低（P<0.05）。结论 NAC可以抑制香烟烟雾提取物造成的A549细胞损伤，IGFBP-3可能在香烟烟雾提取导致细胞损伤的机制中发挥作用。

关键词: IGFBP-3, 香烟烟雾提取物, 肺泡上皮细胞, NAC

Abstract: Objective To study the protective role of N-acetylsteine in the A549 cell injury caused by cigarette smoke exact and the expression change of insulin-like growth factor binding protein -3 in this process. Methods MTT assay was used to evaluate cell viability. Cell cycle and apoptotic cell proportion in each group detected respectively by PI and AnnexinV-FITC/PI double-labelled flow cytometry. Intracellular reactive oxygen species (ROS) was estimated by fluorescent indicator H2DCFDA. DAPI was used to observe the nuclear morphology. Furthermore, using real-time quantitative RT-PCR as well as western blot methods,the expression level of IGFBP-3 was detected.Result Compared with control group(0.78±0.03), CSE inhibit cell proliferation significantly(0.55±0.04). NAC can reduce cell injury caused by CSE including restoring the viability of A549 cells(0.67±0.04), attenuating G1 block of cell cycle and significantly reducing the proportion of apoptotic cells and so on. High expression of insulin-like growth factor binding protein-3 (IGFBP-3) in A549 cells treated with CSE was found at both transcriptional and protein levels, and concomitant with the restoration of cell growth after treatment with NAC, down regulation of IGFBP-3 was observed.Conclusion From the present study, it is concluded that NAC can antagonize CSE-induced growth arrest of alveolar epithelial cells and that down regulation of IGFBP-3 probably play an important role in this process.

Key words: IGFBP-3, cigarette smoke extract, alveolar epithelial cells, NAC

中图分类号:

R365

张强弩焦宗宪常嘉琛. N-乙酰半胱氨酸减轻香烟烟雾提取物所致的A549细胞损伤[J]. 基础医学与临床, 2014, 34(1): 72-77.