天津医药

• 实验研究 • 上一篇    下一篇

氯乙酰胆碱对肠上皮内淋巴细胞产生一氧化氮的影响

孔维涵1,方步武1,吴咸中2   

  1. 1. 天津市天津医院
    2. 天津市中西医结合急腹症研究所
  • 收稿日期:2012-07-11 修回日期:2012-12-08 出版日期:2013-04-15 发布日期:2013-04-15
  • 通讯作者: 孔维涵

Effects of Acetylcholine Chloride on Nitric Oxide Produced by IELs of Mice

KONG Wei han1,FANG Bu wu2,WU Xian zhong3   

  1. 1. Tianjin Hospital, Tianjin 300211, China
    2. Tianjin hospital
    3. Institute of Integrative Medicine Therapy for Acute Abdominal Diseases of Tianjin
  • Received:2012-07-11 Revised:2012-12-08 Published:2013-04-15 Online:2013-04-15
  • Contact: KONG Wei han

PDF (PC)

2495

摘要： 【摘要】目的 探讨氯乙酰胆碱对小鼠肠上皮内淋巴细胞(IELs)产生一氧化氮(NO)的影响。方法 分离、培养 IELs，以不同浓度氯乙酰胆碱作用于IELs，以植物血凝素(PHA)为对照，用荧光分光光度法测定培养上清液中NO，用 MTT分析细胞增殖，以荧光素钙螯合剂（Fura-2/AM）检测细胞内钙([Ca2+]i)。结果与IELs对照组比较(培养液含终浓度0.5 mg/LCon A)，氯乙酰胆碱10-5、10-6、10-7 mol/L组吸光度（A）值均降低(P < 0.05)。IELs培养第4天时NO水平最低 (培养液不含Con A)，此后NO呈进行性升高，至第7天时(观察期末)达峰值且分别高于第1、2、3、4天(P < 0.05)。与IELs 对照组比较，10-5 mol/L氯乙酰胆碱组及10-2 g/L PHA组NO水平减低(P < 0.05)，氯乙酰胆碱组[Ca2+]i水平的差异无统计学意义(P > 0.05)，而PHA 组[Ca2+]i 水平升高(P < 0.05)。氯乙酰胆碱组NO 水平与lg[Ca2+]i 呈正相关（r=0.840，P = 0.036)。结论 氯乙酰胆碱具有抑制IELs产生NO的作用，提示迷走神经调控炎症反应可能与其抑制NO的产生有关。

关键词: 肠上皮内淋巴细胞, 一氧化氮, 氯乙酰胆碱

Abstract:

[Abstract] Objective  To explore effects of acetylcholine chloride on nitric oxide (NO) produced by intestinal intraepithelial lymphocytes (IELs) in mice. Methods  IELs of Balb/c mice were isolated, cultured and incubated with acetylcholine chloride of various concentrations. Phytohemagglutinin (PHA) served as the control. NO in culture supernatants was detected with fluorospectrophotometry, IELs proliferation was determined by MTT, and intracellular Ca2+ ([Ca2+]i) was assayed by Fura-2/AM. Results  Compared with the control group of IELs (with concanavalin A (Con A) of 0.5 mg/L final concentration in the medium), absorbance (A) value decreased in each group of acetylcholine chloride 10-5, 10-6 and 10-7mol/L (P < 0.05). The level of NO in IELs supernatant was the lowest (without Con A in medium) on day 4, but the level of NO was then in? creased progressively and got the peak at day 7 (at the end of observation), which was higher than that of day 1, day 2, day 3 and day 4 (P < 0.05). In comparison with the control group of IELs, the level of NO in 10-5 mol/L acetylcholine chloride group and 10-2 g/L PHA group were significantly lower respectively (P < 0.05). There was no significant difference in the [Ca2+]i level in acetylcholine chloride group (P > 0.05), but it increased significantly in PHA group (P < 0.05). The level of NO in supernatants was positively correlated with common logarithm of [Ca2+]i of IELs only in acetylcholine chloride group (r=0.840，P = 0.036). Conclusion  Acetylcholine chloride can inhibit NO produced by IELs, which suggests that vagus nerve’s effect on controlling inflammation was possibly related to its ability to inhibit the production of NO.

Key words: intestinal intraepithelial lymphocytes, nitric oxide, acetylcholine chloride