GLP-1R过表达对COPD患者气道平滑肌细胞增殖和迁移的影响

摘要

目的:探讨胰高血糖素样肽-1受体(GLP-1R)过表达对慢性阻塞性肺病(COPD)患者气道平滑肌细胞(ASMCs)增殖及迁移的影响.方法:分离培养来自15例COPD患者(观察组)和6例无COPD病史肺部良性肿瘤患者(对照组)的ASMCs.利用PcDNA3.1(空载体组)和PcDNA3.1-GLP-1R(过表达组)质粒转染ASMCs,同时设置空白对照组.分别采用qPCR和westemblotting法检测GLP-1R表达水平,MTT法和Transwell实验检测ASMCs增殖和迁移能力,酶联免疫吸附试验(ELISA)法检测粒细胞-巨噬细胞集落刺激因子(GM-CSF)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-8水平.结果:观察组GLP-1R mRNA和蛋白表达水平明显低于对照组(P＜0.05),过表达组GLP-1R mRNA及蛋白表达水平明显高于空载体组及空白对照组(P＜0.05).过表达组ASMCs增殖和迁移能力明显低于空载体组、空白对照组(P＜0.05),GM-CSF、TNF-α、IL-6、IL-8水平明显亦低于空载体组和空白对照组(P＜0.05).空载体组与空白对照组上述各指标比较,差异均无统计学意义(均P＞0.05).结论:GLP-1R过表达可明显抑制COPD气道平滑肌细胞增殖、迁移,抑制促炎因子释放.%Objective:To investigate the effects of GLP-1R overexpression on proliferation and migration of airway smooth muscle cells (ASMCs)in patients with COPD.Methods:ASMCs were collected from 15 patients with COPD (observation group)and 6 patients with benign lung cancer without COPD (control group).The ASMCs were transfected with PcDNA3.1 and PcDNA3.1-GLP-1R plasmids,and blank control group was set at the same time.The expression levels of GLP-1R were detected by qPCR and western blotting,respectively.The proliferation of ASMCs was detected by MTT assay.The migration ability of cells was detected by trans-well assay.The levels of granulocyte macrophage-colony stimulating factor(GM-CSF),tumor necrosis factor (TNF)-α,interleukin (IL)-6 and IL-8 were detected by enzyme-linked immuno sorbent assay (ELISA).Results:The expressions of GLP-1R mRNA and protein in the observation group was significantly lower than that in the control group (P＜ 0.05).The GLP-1R mRNA and protein levels were significantly increased in overexpression group,compared with the empty vector group and blank control group (P＜ 0.05).The proliferation and migration abilities of ASMCs in overexpression group were significantly lower than those in empty vector group and blank control group (P＜0.05).The levels of GM-CSF,TNF-α,IL-6 and IL-8 in overexpression group were significantly lower than those in empty vector group and blank control group(P＜0.05).There were no significant differences in above indexes between the empty vector group and the blank control group (P＞0.05).Conclusion:Overexpression of GLP-1R could significantly inhibit the proliferation and migration of ASMCs in COPD patients,and suppress the release of proinflammatory cytokines.