氯乙酰胆碱对肠上皮内淋巴细胞产生一氧化氮的影响  

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

作　　者：孔维涵[1] 方步武[2] 吴咸中[3] 

机构地区：[1]天津市天津医院,300211 [2]天津医科大学药理学教研室 [3]天津市中西医结合急腹症研究所

出　　处：《天津医药》2013年第4期353-356,I0002,共5页Tianjin Medical Journal

基　　金：天津市高等学校科技发展基金项目(项目编号:2001YY08);天津医科大学科学基金项目(项目编号:2000KY31)

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0.5mg/L ConA),氯乙酰胆碱10-5、10-6、10-7mol/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ConA),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,10-5mol/L氯乙酰胆碱组及10-2g/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。Objective To explore effects of acetylcholine chloride on nitric oxide （NO） produced by intestinal intraepithelial lymphocytes （IELs） in mice. Methods IELs of Balb/e 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 Fu- ra-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）, absorhance （A） value decreased in each group of acetylcholine chloride 10-5, 10.6 and 10-mol/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 increased 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-s 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 lev-el 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.

关 键 词：一氧化氮 钙 肠上皮内淋巴细胞 氯乙酰胆碱 

分 类 号：R392[医药卫生—免疫学]