摘要
目的探讨普拉梭菌(Faecalibacterium prausnitzii,F.prausnitzii)及其上清液对葡聚糖酸钠(DSS)诱导的结肠炎小鼠Treg/Th17平衡的影响。方法将40只雄性C57BL/6J小鼠随机分为正常组、结肠炎对照组、F.prausnitzii活菌干预组、F.prausnitzii上清干预组,每组10只,正常组正常饮水,其余各组均饮用质量浓度为25 g/L的葡聚糖硫酸钠(DSS)诱导结肠炎模型,同时分别用磷酸缓冲盐溶液(PBS)、F.prausnitzii、F.prausnitzii上清灌胃。流式细胞术检测各组小鼠脾脏中的Treg、Th17比例,并计算各组Treg/Th17比值;ELISA法检测外周血清中IL-10、IL-17水平;RT-PCR检测结肠组织中Foxp3、ROR-γt mRNA的表达。结果与结肠炎对照组相比,F.prausnitzii活菌干预组和F.prausnitzii上清干预组的Treg细胞比例上升,Th17细胞比例下降,Treg/Th17比值升高;且血清中IL-10水平升高,IL-17水平降低(P<0.05)。与结肠炎对照组相比,F.prausnitzii上清干预组Foxp3 mRNA表达上升(P<0.05),但F.prausnitzii活菌干预组Foxp3 mRNA表达差异无统计学意义(P>0.05);两干预组肠组织中ROR-γt mRNA表达较结肠炎对照组下降(P<0.05)。结论 F.prausnitzii及其上清液可调控T细胞亚群,其机制可能为通过提高Foxp3 mRNA表达,抑制ROR-γt mRNA的表达调节Treg/Th17的平衡。
Objective To investigate the effect of Faecalibacterium prausnitzii （F. prausnitzii） and its supernatant on the balance of Treg/Thl7 in mice induced by dextran sulfate sodium. Methods A total of 40 C57BL/6J mice were randomly divided into four groups： normal group, colitis model group, F. prausnitzii treatment group and F. prausnitzii supernatant treatment group, respectively. The normal group drunk water normally, colitis were induced by drinking 25 g/L DSS solution in the other groups, which were injected the PBS, the F. prausnitzii and the F. prausnitzii supernatant through the stomach, respectively. Percentages of Treg cells and Thl7 ceils in spleen were analyzed by flow cytometry, and the ratio of Treg/Thl7 was calculated. Serum levels of IL-10 and IL-17 were measured by ELISA and the levels of Foxp3 and ROR-γt mRNA in colonic tissue were detected by RT-PCR. Results Percentage of Treg cells in F. prausnitzii treatment group and F. prausnitzii supernatant treatment group in spleen was higher than that in colitis model group, and the percentage of Thl7 cells in the two treatment groups in spleen was lower than that in colitis model group. The ratio of Ttreg/Thl7 in the two treatment groups was higher than that in the colitis model group. Serum level of IL-10 in F. prausnitzii treatment group and the F. prausnitzii supernatant treatment group was higher than that in colitis model group, and serum level of IL-17 in the two treatment groups was lower than that in colitis model group. Compared with the colitis model group, the expression of Foxp3 mRNA in F. prausnitzii supernatant treatment group in colonic tissues was increased, but the expression of Foxp3 mRNA had no difference in F. prausnitzii treatment group. The expression of ROR-γt mRNA in the two treatment groups in colonic tissues were lower than that in colitis model group. Conclusion The F. prausnitzii and its supernatant could regulate the T lymphocyte subsets by increasing the expression of Foxp3 mRNA and inhibiting the expression of ROR-γt mRNA, which might regulate the balance of Treg/Th17.
出处
《胃肠病学和肝病学杂志》
CAS
2017年第12期1396-1400,共5页
Chinese Journal of Gastroenterology and Hepatology
基金
国家自然科学基金项目(81470819)
