大承气颗粒剂量和药物血清作用时间对IELs生成一氧化氮的影响

Effects of Dosage of Dachengqi(大承气)Granules and Time Incubated with Sera Containing Drug on NO Produced by Intestinal Intraepithelial Lymphocytes of Mice

目的:探讨大承气颗粒剂量和药物血清作用时间对小鼠肠上皮内淋巴细胞(intestinal intraepithelial lymphocytes,IELs)产生一氧化氮(nitric oxide,NO)的影响。方法:分离、培养、鉴定小鼠IELs;以不同剂量(1.204g/kg、0.602g/kg、0.120g/kg)的大承气颗粒灌胃大鼠后不同时间点的药物血清作用于IELs,另以临床等效剂量(0.602g/kg)大承气颗粒及以大黄酚消除半衰期为时间间隔连续6个t1/2灌胃大鼠的药物血清作用于IELs动态观察7d;用荧光分光光度法测定培养上清液中NO。结果:大承气颗粒多次灌胃的药物血清能持续抑制IELs产生NO,对IELs加药物血清培养1、5、7d产生的NO分别显著低于加培养液培养相应时间产生的NO;加药物血清培养3、5、6、7d产生的NO分别显著低于加正常血清培养相应时间产生的NO(P<0.01,P<0.05)。单次灌胃不同剂量的大承气颗粒后不同时间点的药物血清多可抑制IELs产生NO,其中大承气颗粒0.120g/kg灌胃后12h以及0.602g/kg灌胃后12h和24h的药物血清使IELs产生的NO水平显著低于正常血清组(P<0.05)。结论:大承气颗粒可确切抑制IELs产生NO,以减轻肠道和全身炎症反应、改善MODS患者的肠道免疫功能。

Objective： To explore the effects of dosage of Dachengqi（大承气） granules and time incubated with sera containing drug on nitric oxide（NO） produced by intestinal intraepithelial lymphocytes（IELs） of mice. Methods IELs of Balb/c mice were isolated, cultured and incubated with drug sera from varying time points after different doses （1.204 g/kg,0.602 g/kg and 0.120 g/kg） of Dachengqi granules administered orally to rats, and incubated （0～8 d） with drug sera from rats administered orally with Dachengqi granules of 0.602 g/kg （clinically equivalent dose） for 7 times according to interval by elimination half life of chrysophanol （t1/2 3.20±0.54 （h） ） in Dachengqi granules. NO in culture supernate was detected by fluorospectrophotometry. Results Drug sera from Dachengqi granulae intragastric administration （ig） to rats for 7 times persisted in inhibition of NO production, NO produced by IELs incubated with drug sera for 1, 5, 7 days was significantly less than that produced by IELs incubated with culture medium control at corresponding time points respectively; NO produced by IELs incubated with drug sera for 3, 5, 6, 7 days was significantly less than that produced by IELs incubated with normal sera （not containing drug） control at corresponding time points respectively （P 0.01,P 0.05）. Drug sera at different time points of single medication of Dachengqi granules most inhibited NO production, NO produced by IELs incubated with drug sera of Dachengqi granules of 0.120 g/kg after 12 h administered and 0.602 g/kg after12 h and 24 h was less than that in normal sera （not containing drug） control group （P 0.05）. Conclusion Sera containing drug from Dachengqi granulaes indeed inhibit NO produced by IELs, thus to mitigate intestinal and systemic inflammatroy reactions,aud to improve intestinal immune function of patients with multiple organ disfunction.