摘要
目的 探讨自主神经控制对人肝细胞增殖的影响及其机理 ,并对增殖相关受体定位。方法 用去甲肾上腺素 (NE)及其拮抗剂及激动剂和乙酰胆碱 (Ach)及其阻滞剂作用于人传代正常肝L0 2细胞及肝癌Bel 740 2细胞 ,分别于 4h、2 4h、48h及 72h用改良MTT法检测上述药物对细胞增殖的影响 ;免疫组织化学染色法对肝L0 2细胞的肾上腺素受体 (α1B AR及 β2 AR)和表皮生长因子受体 (EGFR)定位。结果 NE对人正常肝细胞及肝癌细胞具有促增殖作用 ,且随浓度增加其促增殖作用也增加 ,4h的增殖率较其它时间点显著增高 (P<0 .0 5 ) ;NE +心得安组的作用与NE组间差异无显著性意义 ;二羟苯基异丙氨基乙醇的作用不明显 ;Ach对人正常肝细胞的增殖具有抑制作用 ,且随浓度增加其抑制作用也增加 ,加用阿托品在 2 4h及 48h与Ach比抑制率显著降低 (P<0 .0 5 ) ;琥珀胆碱组在 2 4h、48h及 72h对细胞的增殖也具有抑制作用 (P<0 .0 5 ,P<0 .0 1)。所有人肝L0 2细胞都呈α1B AR、β2 AR及EGFR免疫反应阳性 ,主要定位于胞膜及胞浆。结论 血清存在下 ,交感神经递质NE早期对人正常肝细胞L0 2及人肝癌细胞Bel 740 2具有促增殖作用 ,其作用是通过激动α AR实现的 ,β AR对人肝细胞增殖无影响。副交感神经递质Ach对人正常肝细胞L0 2的增殖?
Objective To explore the effects and mechanism of autonomic nervous control on the proliferation of human hepatocytes. And to examine the cellular localization of some related receptors expression in human hepatocytes. Methods Norepinephrine (NE), and its agonist, antagonist, acetylcholine (Ach), and its antagonist have been added to human hepatocyte line L02 and hepatoma cell line Bel 7402. Modified MTT assay was employed to test the effects of them on the proliferation of the two cell lines at 4 h, 24 h, 48 h and 72 h. Immuocytochemical staining was used to examine the cellular localization of alpha1B adrenoceptor (α 1B AR), β 2 AR and epidermal growth factor receptor (EGFR) expression in human hepatocyte line L02. Results NE potentiated the proliferation of human hepatocyte and hepatoma cell, which was enhanced significantly with dose increased. The proliferative rate of 4 h were higher than that of the other time points ( P <0.05). There were no significant differences between the group of NE combined with propanolol and the group of NE alone. Metaproterenol had no significant effect. Ach significantly inhibited the proliferation of human hepatocyte. Its effect was enhanced with dose increased. Atropine significantly attenuated the inhibitory effect of Ach at 24 h and 48 h ( P <0.05) . Scoline alone inhibited hepatocyte proliferation at 24 h, 48 h and 72 h ( P <0.05, P <0.01). In immunocytochemical staining, there were positive responses to α 1B AR, β 2 AR and EGFR in all cultures. It was observed that the responses to α 1B AR, β 2 AR and EGFR were mainly both cytoplasmic and cell membrane localized. Conclusion NE, the sympathetic neurotransmitter, acts via α 1B AR potentiate the proliferation of human hepatocyte and hepatoma cell in the presence of serum. Ach, the vagus neurotransmitter acting via mAchR and nAchR inhibits hepatocyte proliferation.
出处
《中国普外基础与临床杂志》
CAS
2002年第4期238-241,共4页
Chinese Journal of Bases and Clinics In General Surgery
基金
国家自然科学基金主任基金 (项目编号 :39840 0 2 0 )
