高浓度胆汁诱导单核细胞前炎症反应

Induction of Monocyte Proinflammatory Response by High Concentration Bile

目的探讨高浓度胆汁在触发早期炎症反应中的作用。方法 20只Wistar大鼠随机分为体外培养组和阻塞性黄疸(阻黄)体内培养组,体外组为正常大鼠,阻黄体内组开腹结扎切断胆总管。明胶法提取外周血单核细胞(PBMCs)体外培养。体外组的PBMCs体外与不同浓度的胆汁共同培养,分别于1、6、24、48 h后用Western blot和酶联免疫吸附试验(ELISA)检测培养上清液内前炎症因子IL-1β和前IL-1β表达的变化,并与加入细菌内毒素(LPS)的PBMCs相比较。阻黄体内组于术后1、6、24、48、168 h提取PBMCs同法检测两者的变化。结果阻黄动物术后1 h和6 h提取的PBMCs内可见IL-1β和前IL-1β表达,其中术后1 h和6 h前IL-1β的浓度分别为(834.0±111.1)pg/mL和(785.0±102.2)pg/mL,IL-1β的浓度是(681.0±82.5)pg/mL和(602.0±78.2)pg/mL,较其它时间段明显增高(P<0.05)。正常大鼠来源的PBMCs体外培养在高浓度胆汁(5%)刺激下早期(1 h)可以检测到IL-1β(612.0±70.5)pg/mL和前IL-1β(467.0±63.2)pg/mL表达增高(均P<0.05)。同时与LPS共同培养,5%胆汁刺激后1 h、6 h及2%、1%胆汁刺激1 h后可见IL-1β和前IL-1β表达增加。结论进入血循环的高浓度胆汁能触发早期炎症反应,合并细菌感染后作用更强。

Objective To investigate the effects and mechanisms of high concentration bile on monocyte proinflammatory response.Methods Twenty Wistar rats were randomly divided into two groups equally：cell culture in vitro group and in vivo group.The normal rats served as in vitro group,and the animals subject to abdominal incision and ligation of common bile duct served as obstructive jaundice（OJ）in vivo group.The peripheral blood monouclear cells（PBMCs）of rats were isolated and purified by gelatin method,and cultured in vitro.PBMCs from in vitro group were co-cultured with different concentrations of bile,and at 1,6,24,48 h,by using Western blot and ELISA,the changes of pro-inflammatory factors pro-IL-1β and cleavage fragment IL-1β in the culture supernatant were detected and compared with those of PBMCs in the presence of endotoxin（LPS）.The changes of pro-IL-1β and IL-1β in PBMCs in OJ in vivo group were also examined.Results Both pro-IL-1β and IL-1β could be detected after operation.In OJ in vivo group the concentrations of pro-IL-1β at 1st and 6th h were（834.0±111.1） and（785.0±102.2） pg/mL,and those of IL-1β were（681.0±82.5） and（602.0±78.2） pg/mL respectively,which were significantly higher than those at other time points（all P〈0.05）.In the culture supernatant of in vitro group,the pro-IL-1β and IL-1β concentrations were increased to（467.0±63.2） and（612.0±70.5） pg/mL 1 h after stimulation of PBMCs with high concentration of bile（5%）（P〈0.05）.After co-culture with bile in the presence of LPS,pro-IL-1β and IL-1β concentrations were increased 1 and 6 h after stimulation with 5% bile,and 1 h after stimulation with 2% and 1% bile.Conclusion The high concentration of bile in the blood circulation induces the release of IL-1β from mononuclear cells,and triggers the proinflammatory response.The effects are stronger after combination of bacterial infection.