益生菌Faecalibacterium prausnitzii联合溃克灵对大鼠结肠炎的预防作用

The preventive effect of Faecalibacterium prausnitzii combined with Kuikeling on colitis of rats

目的 :探讨益生菌Faecalibacterium prausnitzii(F.prausnitzii)联合溃克灵防治炎症性肠病的疗效及作用机制。方法 :将50只Sprague-Dawley大鼠随机分为5组:正常组、模型组、F.prausnitzii组、溃克灵组、F.prausnitzii联合溃克灵组。造模前各组灌胃1周(正常组和模型组予生理盐水,其余3组给予相应的F.prausnitzii和溃克灵处理)。采用2,4,6-三硝基苯磺酸(2,4,6-trinitrobenzene sulfonic acid,TNBS)法(80 mg/kg TNBS联合等体积无水乙醇)灌肠复制出结肠炎大鼠模型,3 d后全部处死,观察结肠组织损伤情况,行结肠组织病理学评分;ELISA法测定各组大鼠血浆白介素(interleukin,IL)-10、IL-12、肿瘤坏死因子(tumor necrosis factor,TNF)-α、干扰素(interferon,IFN)-γ水平,计算IL-10/IL-12比值;免疫组化检测肠黏膜IL-10及TNF-α的表达。结果 :F.prausnitzii联合溃克灵组与模型组相比,结肠组织损伤明显改善,组织病理学评分显著降低(P<0.05),血浆IL-10、IL-10/IL-12比值显著升高(P均<0.05),血浆IL-12、IFN-γ显著下降(P<0.05,P<0.001),血浆TNF-α下降但差异无统计学意义,肠黏膜IL-10表达明显升高(P<0.01),肠黏膜TNF-α表达明显降低(P<0.05)。与F.prausnitzii组、溃克灵组相比,F.prausnitzii联合溃克灵组大鼠结肠组织病理学评分降低,IL-10/IL-12比值及血浆IL-10、肠黏膜IL-10表达升高,血浆IL-12、IFN-γ、TNF-α及肠黏膜TNF-α表达下降,但差异无统计学意义。结论:益生菌F.prausnitzii联合中药溃克灵对结肠炎大鼠肠黏膜的预防保护作用,可能与促进外周血及肠黏膜IL-10分泌从而抑制外周血IL-12、TNF-α、IFN-γ产生、降低肠黏膜TNF-α表达等免疫调节机制有关。

Objective：To study the effects and the mechanism of Faecalibacterium prausnitzii（F.prausnitzii）combined with Kuikeling on colitis of rats with inflammatory bowel disease（IBD）. Methods：A total of 50 rats was randomly divided into 5 groups,which named the normal group,the model group,the Kuikeling group,the F.prausnitzii group and the F.prausnitzii combined Kuikeling group,respectively. Each group was given medicine by gavage for a week before model establishment（the normal and model group received saline,the other groups received corresponding F.prausnitzii and Kuikeling）. Colitis model was established by 2,4,6-trinitrobenzene sulfonic acid（TNBS） method（80 mg / kg TNBS combined with equad volume ethanol）. All rat were killed after 3 days,and mucosal lesion area and colon mucosal inflammation were assessed by the colorectal histological damage scores（HDS）. Levels of interleukin-10（IL-10）,interleukin-12（IL-12）,tumor necrosis factor-a（TNF-α）and interferon-γ were detected by ELISA in the plasma,and then we calculated the ratio of IL-10 / IL-12. The expressions of IL-10 and TNF-α in the colon mucosal tissues were detected by immunohistochemical（IHC） method. Results：Compared with the model group,mucosal lesion area and the colon HDS were significantly decreased（P 〈0.05）,the levels of IL-12 and IFN-γ in the plasma（P 〈0.05,P 〈0.001）and the expression of TNF-α inthe colon mucosal tissues（P 〈0.05） were significantly decreased in F.prausnitzii combined Kuikeling group,and the level of TNF-αin the plasma was decreased without statistical significance,the levels of IL-10 in the plasma and the colon mucosal tissues（P 〈0.05,P 〈0.01）and the ratio of IL-10 / IL-12（P 〈0.05）were significantly increased in the F.prausnitzii combined Kuikeling group. Compared with the Kuikeling group and the F.prausnitzii group,mucosal lesion area and the colon HDS were decreased,the levels of IL-12,IFN-γ,and TNF-α in the plasma and the expression of TNF-α in the colon mucosal tissues were decreased,the levels of IL-10 in the plasma and the colon mucosal tissues,the ratio of IL-10 / IL-12 were increased in the F.prausnitzii combined Kuikeling group,but those differences had no statistical significance. Conclusion：The preventive and therapeutic effects of F.prausnitzii combined Kuikeling is better than single treatment. The immune regulation mechanism may be related to promoting secretion of IL-10 in the plasma and the colon mucosal tissues and decreasing IL-12,TNF-α,IFN-γ levels in the plasma and TNF-α level in the colon mucosal tissues.