益生菌Faecalibacterium prausnitzii联合溃克灵对大鼠结肠炎的预防作用
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国家自然科学基金面上项目(81470819)


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

    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。

    Abstract:

    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 in the colon mucosal tissues (P < 0.05) were significantly decreased in F.prausnitzii combined Kuikeling group,and the level of TNF-α in 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 ofF.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.

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毋攀锋,李 琳,于成功.益生菌Faecalibacterium prausnitzii联合溃克灵对大鼠结肠炎的预防作用[J].南京医科大学学报(自然科学版),2016,(9):1079-1084

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  • 收稿日期:2016-01-29
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  • 在线发布日期: 2016-09-18
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