摘要
目的:探讨成熟树突状细胞(dendritic cells, DCs)是否存在非神经元性胆碱能系统及其调控机制。方法:分离正常小鼠骨髓细胞,体外用细胞因子(IL-4,GM-CSF)诱导、分化为幼稚DCs,细菌脂多糖(LPS)刺激其成熟。光镜观察其形态变化、流式细胞术进行分子鉴定。免疫细胞化学方法、免疫荧光抗体技术、流式细胞术和RT-PCR方法检测DCs烟碱样乙酰胆碱受体亚单位α7(nAChRα7)、胆碱乙酰转移酶(ChAT)和乙酰胆碱酯酶(AChE)。同时,流式细胞术检测美加明(MEC)作用于成熟DCs 12 h前后nAChRα7表达。结果:成熟DCs表达nAChRα7、ChAT、AChE蛋白和mRNA;nAChRα7蛋白主要分布于细胞膜,ChAT和AChE分布于细胞浆。其中AChE蛋白表达强于ChAT(P<0.05),与nAChRα7比较有上升趋势;MEC作用组nAChRα7蛋白表达明显低于无MEC作用组(P<0.05)。结论:成熟DC存在固有胆碱能系统,外源性因素(美加明等)可调节其效应。DCs可能通过胆碱能系统而发挥其神经免疫调节作用。
AIM： To investigate whether there was nicotinic acetylcholine receptor subunit α 7 （nAChR α 7 ）, choline acetyhransferase（ CHAT）, acetylcholinesterase（ACHE） expression and its regulation in mature dendritic cells （DCs）. METHODS： Bone marrow（BM） -derived DCs from healthy BALB/c mice were incubated with rmGM -CSF and rmIL- 4, and stimulated to mature with LPS. -Meanwhile, light microscope and flow cytometry were used to identify DCs, as well as immunocytochemistry, immunofluorescence, flow cytometry and RT- PCR methods were used to dectect expression of nAChR α 7, ChAT and ACHE. Flow cytometry was also used to analyze nAChR α 7 expression with mecamylamine （ MEC ） in 12 h. RESULTS： Both protein and mRNA expression of cholinergic system nAChR α 7, ChAT and AChE were found in mature DCs. Furthermore, nAChR α 7 distributed principally in cell membrane, while ChAT and AChE in cytoplasm. Protein expression of AChE was stronger as compared with ChAT （ P 〈 0.05 ）, and there was a trend toward increasing as compared with nAChR ct 7. And then, the expression of nAChR α 7 was down regulated by MEC as compared with the group without MEC stimulation（ P 〈 0.05 ）. CONCLUSION： An innate cholinergic system was in mature DCs, which was affected by extrinsic factor （i. e. , MEC）. And it may be involved in anti -inflammation immune adjustion of cholinergic closed - circuit.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第3期428-434,共7页
Chinese Journal of Pathophysiology
基金
Supported by the Foundation of National Natural Science(No30170442)
关键词
树突细胞
胆碱能系统
美加明
Dendritic cells
Cholinergic system
Mecamylamine
