摘要
目的探讨尼古丁受体在尼古丁刺激大鼠气道平滑肌细胞(RASMCs)增殖中的作用。方法分别设计相应的RNA引物,应用RT-PCR实验,观察RASMCs上存在的尼古丁受体(n Ach Rs)亚单位情况;应用细胞计数法检测尼古丁受体非特异性拮抗剂MCA和α7尼古丁受体特异性拮抗剂MG624对尼古丁作用的RASMCs增殖作用的影响;EDU掺入法测定应用尼古丁受体拮抗剂MCA和MG624对尼古丁作用的RASMCs DNA复制活性的影响。结果RT-PCR实验证实大鼠气道平滑肌细胞上存在n Ach Rs亚单位,分别是α1、4、7、9、10,β1、2、4。细胞计数法检测发现尼古丁受体拮抗剂MCA、MG624均可以抑制尼古丁的促增殖作用,细胞数量明显下降,与尼古丁单独作用组比较差异有统计学意义(P<0.05);EDU检测显示尼古丁受体拮抗剂MCA和MG624均可以抑制尼古丁的作用,使DNA复制活性降低,细胞复制水平下降,与尼古丁单独作用组比较差异有统计学意义(P<0.05)。结论尼古丁能够通过尼古丁受体介导促进大鼠气道平滑肌细胞的增殖。
Objective To study the effects of nicotinic acetylcholine receptors（n Ach Rs） antagonists on the nicotineinduced proliferation of rat airway smooth muscle cells（RASMCs）. Methods Subunit-specific primers for n Ach Rs were designed for the amplification of unique regions of each subunit. Reverse-transcription polymerase chain reaction（RTPCR） was performed to examine the n Ach R present in RASMCs. Cell numbers were counted with a Zeiss Coulter Counter to detect the effects of n Ach Rs antagonist MCA and MG624 on the nicotine-induced proliferation of RASMCs.EDU incorporation assay was carried out to detect the effects of n Ach Rs antagonists MCA and MG624 on the cells replication.Results We found that eight different n Ach R subunits were expressed in RASMCs, that is α1, α4, α7, α9, α10, β1,β2 and β4. The nicotine-induced proliferation of the RASMCs was significantly inhibited by the n Ach R antagonists MCA and MG624; the cell number decreased significantly compared to 10 μmol / L nicotine group（P 〈0.05）. The EDU incorporation assays showed that the nicotine-induced proliferation of the RASMCs was significantly inhibited by the n Ach R antagonists MCA and MG624; the level of cells replication decreased significantly compared to 10 μmol / L nicotine group（P〈0.05）. Conclusion Nicotine can induce the proliferation of RASMCs via nicotinic acetylcholine receptors（n Ach R）.
出处
《热带医学杂志》
CAS
2014年第12期1541-1544,F0004,共5页
Journal of Tropical Medicine
基金
国家自然科学基金(81170043)
广州医科大学博士启动基金(2013C09)
