中药对湿热型溃疡性结肠炎小鼠NFκB p65及免疫功能的影响

Effect of Herbal Decoction on NFκB p65 and Immune Function in the Ulcerative Colitis Mice with Dampness Heat Syndrome

目的:探讨清热利湿方对湿热型溃疡性结肠炎(ulcerative colitis,UC)小鼠不同时段结肠黏膜核转录因子κB(NFκB)p65蛋白表达及相关免疫功能的影响。方法:选用80只SPF级雄性Balb/c小鼠,随机分为对照组、造模组、西药组、中药组,每组20只,除对照组外,其余3组采用2,4,6-三硝基苯磺酸(TNBS)/乙醇溶液灌肠法配合饮食加环境复合法建立小鼠湿热型UC模型,术后西药组予以美沙拉嗪灌肠,中药组予以清热利湿方灌肠;第3 d、7 d每组各取10只小鼠观察血清肿瘤坏死因子-α(TNF-α)、IgA、黏膜sIgA水平,结肠黏膜NFκB p65蛋白表达情况。结果:造模组TNF-α、IgA、黏膜sIgA水平升高,NFκB p65蛋白表达增加,与对照组有显著差异(P<0.01);与造模组相比,中药组、西药组TNF-α、IgA、黏膜sIgA、NFκB p65蛋白表达下降(P<0.01);中药组与西药组相比,给药3 d,上述各指标无明显差异(P>0.05)。给药7 d,中药组TNF-α为(240.16±16.24)pg/mL、IgA为(161.46±28.75)ng/mL、黏膜sIgA为(18.25±2.20)μg/mL、NFκB p65蛋白表达阳性率为33.3%,较西药组TNF-α(290.58±18.33)pg/mL、IgA(184.90±33.23)ng/mL、黏膜sIgA(20.86±1.84)μg/mL、NFκB p65蛋白表达阳性率(55.6%)明显降低(P<0.05)。结论:湿热型UC动物模型造模成功,清热利湿方治疗湿热型UC疗效肯定,作用机制可能与抑制NFκB p65蛋白表达、降低TNF-α、IgA、黏膜sIgA水平等相关免疫指标有关,随疗程增加,中药疗效优于西药。

Objective To investigate the effect on the protein expression of NFKB p65 and immune of clearing heat and removing dampness herbal decoction function in the colon tissue of mice with ulcerative colitis with dampness heat syndrome. Methods Eighty male Balb/c mice of SPF grade with 6-8 weeks old were ran- domly divided into control group, model group, Western medicine group and Chinese medicine group, with 20 mice in each group. Except for the control group, the remaining groups were given 2, 4, 6-trinitrobenzene sulfon- ic acid （TNBS） and ethanol enema combined with damp-heat type diet and environmental composite method to establish mice UC model. Western medicine group were administered with Mesalazine at the daily dose of 0.22 g/ kg by enema 1 mL BID; Chinese medicine group were administered with herbal decoction at the daily dose of 32 g/kg by enema 1 mL BID. Serum TNF-et, IgA, mucosal sIgA levels, NFKB p65 protein expression were de- tected in 10 mice of each group on day 3 and day 7. Results TNF-α, IgA, mucosal sIgA levels and NFKB p65 protein expression in model group were increased significantly than that in the control group （P 〈 0.01）. Compared with the model group, the expression of TNF-α, IgA, mucosal sIgA, and NFKB p65 protein were de- creased in Western medicine group and Chinese medicine group. There was no difference for above parameters between Western medicine group and Chinese medicine group after 3 days treatment. In Chinese medicine group after 7 days treatmen, TNF-α （240.16± 16.24） pg/ mL, IgA （161.46 ± 28.75） ng/mL, mucosal slgA （18.25±2.20） μg/mL and NFKB p65 protein expres-sion （33.3%） were significantly lower than those in Western medicine group with TNF-α（290.58±18.33） pg/mL, IGA（184.90±33.23） ng/mL, mucosal slgA（20.86±1.84） μg/mL and NFKB p65 protein expression （55.6%）. There was significant difference between two groups（P〈 0.05）. Conclusion The UC mice model with dampness heat syndrome were successfully modeled. The effect of clearing heat and removing dampness herbal decoction in the treatment of damp-heat type UC was obvious. The mechanisms could be the inhibition of NFKB p65 pro- tein and decrease in the related immune indexes of TNF-α, IgA and mucosal slgA levels. With the course of treatment increased, the curative effect of traditional Chinese medicine is better than that of Western medicine.