Article Summary
任安民,吴宏宪,郭真真,操群安,戴秋艳,刘丽萍.nAChRα1调控尼古丁促进巨噬细胞RAW264.7增殖迁移的作用研究[J].现代生物医学进展英文版,2018,(8):1474-1478.
nAChRα1调控尼古丁促进巨噬细胞RAW264.7增殖迁移的作用研究
nAChRα1 Mediates Nicotine-induced the Proliferation and Migration of RAW264.7
Received:February 14, 2018  Revised:February 28, 2018
DOI:10.13241/j.cnki.pmb.2018.08.014
中文关键词: nAChRα1  尼古丁  细胞增殖  细胞迁移
英文关键词: nAChRα1  Nicotine  Cell proliferation  Cell migration
基金项目:国家自然科学基金项目(81670399)
Author NameAffiliationE-mail
任安民 上海交通大学附属上海市第一人民医院心内科 上海 200080 renanmin2016@163.com 
吴宏宪 上海交通大学附属上海市第一人民医院心内科 上海 200080  
郭真真 上海交通大学附属上海市第一人民医院心内科 上海 200080  
操群安 上海交通大学附属上海市第一人民医院心内科 上海 200080  
戴秋艳 上海交通大学附属上海市第一人民医院心内科 上海 200080  
刘丽萍 盐城市第一人民医院心内科 江苏 盐城 224000  
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中文摘要:
      摘要 目的:探讨nAChRα1是否参与调节尼古丁促进巨噬细胞RAW264.7增殖迁移的作用。方法:将体外培养的RAW264.7细胞分4组为:①正常对照组;②尼古丁组;③对照干扰+尼古丁组;④nAChRα1干扰+尼古丁组。用尼古丁(5 ng/mL)刺激巨噬细胞RAW264.7,特异性nAChRα1 siRNA用脂质体3000转染细胞,CCK-8法检测尼古丁处理3 h、24 h和48 h后细胞的增殖情况,细胞划痕实验检测细胞迁移情况,Western blot和RT-PCR检测细胞内nAChRα1、MMP-2、MMP-9的蛋白和mRNA的表达情况。结果:与空白对照组相比,尼古丁可显著促进RAW264.7细胞的增殖和迁移,增加nAChRα1、MMP-2、MMP-9的蛋白和mRNA表达;而在干扰nAChRα1表达后,尼古丁诱导的RAW264.7细胞的增殖和迁移明显被抑制,且细胞nAChRα1、MMP-2、MMP-9的蛋白和mRNA表达均显著的降低。结论:nAChRα1可介导尼古丁促进RAW264.7细胞的增殖和迁移,这可能与其参与调控尼古丁增加RAW264.7细胞分泌MMP-2、MMP-9有关。
英文摘要:
      ABSTRACT Objective: To investigate whether nAChRα1 mediate nicotine-induced proliferation and migration of macrophages RAW264.7. Methods: RAW264.7 cells cultured in vitro were divided into four groups: ① Control group;②Control+Nicotine group;③NC-siRNA+Nicotine group; ④α1-siRNA+Nicotine group. RAW264.7 Cells were treated with nicotine(5 ng/mL), nAChRα1-siRNA was transfected into RAW264.7 cells by Lipofectamine 3000 reagent; The cell proliferation at 3 h, 24 h and 48 h after nicotine treatment was measured by CCK-8 assay; Scratch wound healing was used to measure the migration of cells. The protein and mRNA expression of nAChRα1, MMP-2, MMP-9 were determined by Western blot and RT-PCR. Results: Compared with the control group, the proliferation and migration of RAW264.7 cells under nicotine exposure were significantly increased(P<0.05). In addition, the protein and mRNA expression of nAChRα1, MMP-2, MMP-9 were obviously increased(P<0.05). However, the inhibitor of nAChRα1 by si-RNA interference can significantly reduce the proliferation and migration of RAW264.7 cells under nicotine exposure. And the protein and mRNA expression of nAChRα1, MMP-2, MMP-9 were significantly decreased(P<0.05). Conclusion: nAChRα1 mediated nicotine-induced proliferation and migration of RAW264.7 cells, which may related to the promotion of MMP-2 and MMP-9 secretion.
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