雷公藤多甙对尼古丁预处理的小鼠肠炎模型脾淋巴细胞因子的影响机制

Inhibitory effects of multi glycosidorum triptery on overproduction of lymphokines by splenocytes in nicotine pretreated and oxazolone-induced colitis in mice

目的:观察尼古丁预处理的小鼠肠炎模型体外实验中,雷公藤多甙对脾脏淋巴细胞生成细胞因子的影响,从免疫学、分子生物学角度探讨雷公藤多甙对经尼古丁预处置的炎症性肠病模型的作用,分析雷公藤多甙与尼古丁在溃疡性结肠炎的治疗方面是否有协同作用。 方法:用半抗原物质恶唑酮(oxazolone,以下简称OXZ)诱导SJL/J小鼠肠炎模型(n=7)。OXZ对照组:予6mg溶于乙醇灌肠3d后处死;尼古丁预处置组:在用OXZ灌肠3d处死前,每日予尼古丁0.5mg/kg连续皮下注射3wk的预处置。均收集脾细胞,计数。加入10μg/L PMA(phorbol myristate acetate,Sigma,St.Louis,MO,USA)和100μg/L Ionomycin(Sigma)作24h培养,离心后收集上清液置于-70℃保存,行ELISA检测。MGT实验组:将0.1g/L和0.01g/L浓度的雷公藤多甙(MGT)加入培养的淋巴细胞液中(未加的为空白对照组),用ELISA检测IFN-γ和IL-4的生成量。 结果:与OXZ组相比,尼古丁预处理组的IFN-r显著增高(0.65±0.08 ng/L vs 2.95±0.064 ng/L);随着剂量增加(2.95±0.65ng/L→1.448±0.28 ng/L→0.553 ng/L±0.07 ng/L,P<0.01),MGT明显地抑制尼古丁刺激IFN-γ的过量分泌。随着MGT剂量的增加(6.95±0.29 ng/L→6.90±0.65 ng/L→5.48±0.44 ng/L,P<0.05),尼古丁预处理组的IL-4生成较OXZ组的少(7.

AIM: To investigate the in vitro effect of multi-glycosidorum triptery (MGT) on lymphokine production by splenocytes in nicotine-pretreated and oxazolone (OXZ)-induced colitis of murine model.METHODS: Male SJL/J mice were divided into OXZ group and nicotine pretreated group (NP group), 6 mg of OXZ (in ethanol) was administered intrarectally to induce colitis and the mice were killed 3 days later. For nicotine group, 0.5 mg/kg every day of nicotine was injected subcutaneously for 3 weeks before OXZ was administered. Isolated splenocytes were cultured for 24 hours in the presence of PMA and ionomycin with MGT of 0.1 g/L or 0.01 g/L in the culture medium of splenocytes, and the blank samples were as control. Production of IFN-r and IL-4 in the supernatant wasmeasured by ELISA.RESULTS: Compared with OXZ group, significantly increased IFN-r was noticed (0.65±0.08 ng/L vs 2.95 ±0.64 ng/L) in NP group; MTG significantly inhibited nicotine stimulated overproduction of IFN-r in a dose independent manner (2.95 ±0.65 ng/L→ 1.448 ±0.28 ng/L→ 0.553 ± 0.07 ng/L, P<0. 01). IL-4 production in NP group was lower than that of OXZ group(7.83±0.69 ng/L vs 6.95 ±0.29 ng/L, P< 0.05) and was further decreased! by MTG in a dose dependent manner (6.95 ±0.29ng/L→6.90±0.65ng/L→5.48±0.44ng/L, P<0.05).CONCLUSION: MTG not only suppresses the IFN-y overproduction stimulated by nicotine and also has a cooperative effect with nicotine in terms of inhibition of IL-4 production. MGT may counteract the deteriorating effect of nicotine on Th1 dominant response (INF-r) and have a coordinate effect with nicotine for suppression of Th2 dominant response (IL-4).