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为了解乙酰胆碱 (acetylcholine,ACh)在人垂体腺瘤发生、发展中的作用 ,本研究首先观察了人垂体腺瘤细胞内是否存在合成ACh必需的胆碱乙酰转移酶 ,并应用MTT实验、[3 H]TdR掺入实验、细胞周期分析和TUNEL法测定了ACh对体外培养的人垂体无功能瘤、催乳素瘤和生长激素瘤等三种瘤细胞增殖和凋亡的影响。结果发现 :(1)三种垂体腺瘤细胞中均有胆碱酯酶的表达 ,但明显少于正常垂体 ;(2 )ACh对三种类型的人垂体腺瘤细胞增殖代谢的影响相类似 ,不同浓度的ACh能明显抑制体外培养的三种人垂体腺瘤细胞的增殖 ,呈明显的剂量效应关系 ,同时ACh能减少垂体腺瘤细胞进入S、G2 期的细胞比例 ,而使处于G1期的细胞比例增加 ;(3)ACh的这种作用可被阿托品阻断 ,但不受筒箭毒的影响 ;(4 )ACh对体外培养的三种人垂体腺瘤细胞凋亡无明显影响。该结果提示 ,ACh可能以旁分泌或自分泌的方式作用于垂体前叶细胞 ,对垂体腺瘤细胞增殖分化有调控作用 ,而且是通过AChM受体来实现的。
To understand the role of acetylcholine (ACh) in the development and progression of human pituitary adenomas, the present study first investigated the presence or absence of choline acetyltransferase required for the synthesis of ACh in human pituitary adenoma cells, and applied MTT assay, [3]. H]TdR incorporation assay, cell cycle analysis and TUNEL assay were used to determine the effects of ACh on the proliferation and apoptosis of human non-tumor pituitary tumors, prolactinoma and growth hormone tumors. The results were as follows: (1) The expression of cholinesterase in all three pituitary adenoma cells was significantly lower than that in the normal pituitary; (2) The effect of ACh on the proliferation and metabolism of three types of human pituitary adenoma cells was similar. Different concentrations of ACh significantly inhibited the proliferation of human human pituitary adenoma cells cultured in vitro and showed a significant dose-effect relationship. At the same time, ACh reduced the proportion of pituitary adenoma cells entering the S and G2 phases, leaving G1 at the same time. The proportion of cells increased; (3) this effect of ACh can be blocked by atropine, but not by tuberculosis; (4) ACh has no significant effect on the apoptosis of human pituitary adenoma cells cultured in vitro. The results suggest that ACh may act on the anterior pituitary cells in a paracrine or autocrine manner and have a regulatory effect on the proliferation and differentiation of pituitary adenoma cells, and is achieved through AChM receptors.