一株猪肠道Lactobacillus animalis LGM对结肠炎小鼠Th细胞分化转录因子基因表达的影响
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国家自然科学基金(31430082)


Effects of Lactobacillus animalis LGM on transcription factors gene expressions in DSS-induced colitis mice
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    [目的]从体外和体内研究Lactobacillus animalis LGM对Th细胞分化转录因子T-bet、GATA-3、ROR-γt和Foxp3的调节作用,以及探究L. animalis LGM对小鼠结肠炎的影响。[方法]本试验采用改良型的Hungate滚管技术从猪结肠内容物中分离一株L. animalis LGM,根据其16S rRNA序列进行鉴定。收集L. animalis LGM培养液上清,与细菌脂多糖(LPS,2μg/mL)同时孵育Caco-2细胞24 h,体外研究L. animalis LGM对Caco-2细胞内Th细胞分化转录因子(T-bet,GATA3,ROR-γt和Foxp3) mRNA表达的影响;配制L. animalis LGM细菌悬液,研究L. animalis LGM灌胃对DSS诱导结肠炎小鼠症状及结肠Th细胞分化转录因子和细胞因子(IFN-γ,IL-4,IL-17和IL-10) mRNA表达的影响,表达结果采用荧光定量PCR法检测。[结果]与对照组相比,L. animalis LGM培养液上清显著上调Caco-2细胞内ROR-γt与Foxp3 mRNA表达(P<0.05),显著下调GATA3、IL-4、IL-17和TGF-β mRNA表达(P<0.05)。L. animalis LGM灌胃显著上调小鼠结肠内ROR-γt和Foxp3的表达(P<0.05),显著降低了促炎因子IL-4和IL-17的表达(P<0.05),阻止了小鼠结肠长度缩短(P<0.05)。[结论]猪肠道分离L. animalis LGM表现出对Th细胞分化转录因子的选择性调节,显著上调Caco-2细胞及结肠炎小鼠ROR-γt与Foxp3 mRNA表达。降低DSS诱导结肠炎小鼠炎症水平,对DSS诱导结肠炎起保护作用,有助于维护肠道环境稳态。

    Abstract:

    [Objective] Investigating the relationship between Lactobacillus animalis LGM and transcription factors (T-bet, GATA3, ROR-γt and Foxp3) gene expressions in vivo or in vitro and observing the effects of L. animalis LGM gavage on DSS-induced colitis.[Methods] L. animalis LGM was isolated from pig colon contents using Hungate rolling tube technique. Bacterial supernatants were extracted and incubated Caco-2 cells with LPS (2 μg/mL) simultaneously. Bacterial suspension was adjusted to 1×109 CFU/0.2 mL and gavaged to DSS-induced mice. Gene expressions of transcription factors (T-bet, GATA-3, ROR-γt and Foxp3) and intracellular cytokines (IL-4, IL-17, INF-γ and IL-10) were analyzed by real-time PCR. [Results] In Caco-2 cells culture experiment, compared to control and LPS groups, L. animalis LGM supernatants treatment significantly downregulated the GATA-3, IL-4, IL-17 and TGF-β gene expressions and upregulated Foxp3 and ROR-γt gene expressions (P<0.05). In DSS-induced colitis mice, compared to the DSS group, L. animalis LGM administration restored the colonic length and increased ROR-γt and Foxp3 gene expressions (P<0.05). [Conclusion] L. animalis LGM isolated from pig colon contents in the present study have distinctive effects on transcription factors. ROR-γt and Foxp3 expressions were significantly upregulated both in Caco-2 cells and DSS-induced colitis mice. L. animalis LGM showed protection on DSS-induced colitis and gut inflammation.

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程颍州,慕春龙,朱伟云.一株猪肠道Lactobacillus animalis LGM对结肠炎小鼠Th细胞分化转录因子基因表达的影响.微生物学报,2019,59(9):1765-1777

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  • 收稿日期:2019-02-24
  • 最后修改日期:2019-05-11
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  • 在线发布日期: 2019-08-29
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