| 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].,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 |
| 投稿时间：2018-02-14 修订日期：2018-02-28 |
| 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） |
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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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