大黄牡丹汤提取液对结肠炎小鼠的治疗及作用机制研究

Therapeutic Effect and Mechanism of Alcohol Extract from Dahuang Mudan Decoction on Colitis Mice Induced by Trinitrobenzene Sulfonic Acid

目的研究大黄牡丹汤提取液对2,4,6-三硝基苯磺酸(TNBS)诱导的结肠炎小鼠的治疗作用。方法TNBS/乙醇溶液一次性灌肠建立结肠炎小鼠模型,大黄牡丹汤提取液低剂量(0.5 g.L-1)、中剂量(1 g.L-1)、高剂量(2 g.L-1)3个不同剂量灌胃治疗6 d,通过观察小鼠一般情况及结肠病理学变化,检测大黄牡丹汤提取液对结肠炎小鼠的治疗作用。用流式细胞术检测外周血、派氏结、肠系膜淋巴结中Treg细胞的变化,ELISA检测小鼠外周血白细胞介素10(IL-10)、转化生长因子(TGF-β)的变化。结果与模型组比较,大黄牡丹汤提取液各组小鼠疾病活动指数(DAI)评分、大体评分、病理学评分都有降低趋势。中剂量组小鼠大体评分、病理学评分都明显降低(2.29±0.76,2.86±0.69 VS 5.86±0.90,5.86±1.07,P<0.05),外周血、肠系膜淋巴结、派氏结Treg细胞,外周血IL-10、TGF-β升高,差异有统计学意义(P<0.05)。结论大黄牡丹汤提取液各剂量组对结肠炎小鼠均有一定的治疗作用,治疗的机制可能与提高Treg细胞水平有关。

Objective To study the therapeutic effect of alcohol extract from Dahuang Mudan Decoction （DMD） on experimental mice colitis induced by trinitrobenzene sulfonic acid （TNBS）. Methods The colitis mice model was established by Single-dose enteroclysis of TNBS-alcohol solution. After the modeling, low-dose（0.5 g·L^-1, middle-dose （1 g·L^-1 , and high-dose （2 g·L^-1 DMD alcohol extract were administered to the mice by intragastric garage for 6 days. Through the observation of body weight, stool, colon pathological change, the therapeutic effect of DMD alcohol extract on colitis mice was evaluated. The changes of Treg cells in peripheral blood, Peyer＇s patches（PPs）, mesenteric lymph nodes （MLN） were tested by flow cytometry, and the changes of interleukin-10（IL-10） and transforming growth factor beta（TGF-β） in peripheral blood were tested by enzyme linked immunosorbent assay （ELISA）. Results Compared with the model group, DA! score, general score and pathological score of DMD alcohol extract groups were decreased, and significant differences were shown in the decrease of general score and pathological score of mid dle-dose DMD alcohol extract group（2.29 ± 0.76, 2.86 ± 0.69 VS 5.86 ± 0.90, 5.86 ± 1.07, P 〈 0.05） , and in the increase of Treg cells count in peripheral blood, PPs and MLN, as well as the peripheral IL-10 and TGF-β （P〈 0.05 ）. Conclusion DMD alcohol extract has certain therapeutic effect on mice colitis induced by TNBS, and the therapeutic mechanism may be related with the increase of the level of Treg cells.