α7烟碱样乙酰胆碱受体在重症肌无力患者外周血单个核细胞中的表达及对T细胞活化的影响

Expression ofα7 nicotinic acetylcholine receptor in peripheral blood mononuclear cells of patients with myasthenia gravis and its effects on T cell activation

目的:探讨α7烟碱样乙酰胆碱受体(α7 nAChR)在重症肌无力(MG)患者外周血单个核细胞(PBMC)中的表达及其对T细胞活化的影响。方法:采用qRT-PCR、Western blot法检测健康对照组(n=20)及MG患者组(n=20)PBMC中α7 nAChR的表达情况,用ELISA检测血清IFN-γ、TNF-α、IL-4、IL-6、IL-10和IL-12水平。收集MG患者的PBMC,诱导T细胞活化,将活化的T细胞分为空白对照组、α7 nAChR拮抗剂组、α7 nAChR激动剂组,培养72 h后采用流式细胞术检测T细胞亚群的增殖情况,并用ELISA检测培养液上清中IFN-γ、TNF-α、IL-4、IL-6、IL-10和IL-12水平。结果:与健康对照组相比,MG患者组α7 nAChR mRNA及蛋白表达水平降低,血清IFN-γ、TNF-α、IL-6、IL-12水平升高(P均<0.001)。与空白对照组相比,α7 nAChR拮抗剂组T细胞亚群增殖增强,IFN-γ、TNF-α、IL-6水平升高(P均<0.05),而α7 nAChR激动剂组T细胞亚群增殖受抑,IFN-γ、TNF-α、IL-6水平降低(P均<0.05)。结论:α7 nAChR在MG患者PBMC中的表达降低,α7 nAChR可能参与调控MG的发生。

Aim:To investigate the expression ofα7 nicotinic acetylcholine receptor(α7 nAChR)in peripheral blood mononuclear cells(PBMC)of patients with myasthenia gravis(MG)and its effects on T cell activation.Methods:The expression ofα7 nAChR in PBMC of healthy control group(n=20)and MG patient group(n=20)was detected by qRT-PCR and Western blot,and serum levels of IFN-γ,TNF-α,IL-4,IL-6,IL-10,and IL-12 were measured using ELISA.The T cells from PBMC of MG patients were induced activation and then divided into blank control group,α7 nAChR antagonist group,andα7 nAChR agonist group.After 72 hours culture,the proliferation of T cell subsets in 3 groups was detected by flow cytometry.The levels of IFN-γ,TNF-α,IL-4,IL-6,IL-10 and IL-12 in cultural supernatant were detected by ELISA.Results:Compared with the healthy control group,the expressions ofα7 nAChR mRNA and protein in the MG patient group decreased,and serum levels of IFN-γ,TNF-α,IL-6,and IL-12 increased(P<0.001).Compared with the blank control group,the proliferation of T cell subsets was enhanced in theα7 nAChR antagonist group,and the levels of IFN-γ,TNF-αand IL-6 were increased(P<0.05),however,in theα7 nAChR agonist group,the proliferation of T cell subsets was inhibited,and the levels of IFN-γ,TNF-αand IL-6 decreased(P<0.05).Conclusion:The expression ofα7 nAChR is reduced in PBMC of MG patients,which indicates thatα7 nAChR may be involved in regulating the occurrence of MG.