酪酸梭菌、蒙脱石散、美沙拉秦对UC大鼠Treg和Th17的影响

Impacts of Clostridium,Montmorillonite Powder and Mesalazine on Treg and Th17 in Rats with UC

目的初步研究酪酸梭菌、蒙脱石散、美沙拉秦对2,4,6三硝基苯磺酸(TNBS)/乙醇法诱导的溃疡性结肠炎(ulcerative colitis,UC)大鼠调节性T细胞(Treg)/辅助性T细胞17(Th17)免疫平衡的影响.方法 48只大鼠随机分为模型组、酪酸梭菌组、蒙脱石散组、美沙拉秦组、美沙拉秦联合蒙脱石散组、正常组,共6组,每组8只大鼠.通过TNBS/乙醇法建立溃疡性结肠炎大鼠的模型.按大鼠体表面积折算药量,每日灌服相应剂量的生理盐水、酪酸梭菌、蒙脱石散、美沙拉秦、美沙拉秦联合蒙脱石散治疗.第5天处死每组4只大鼠,第12天后处死余下的全部大鼠,收集大鼠的血标本.采用流式细胞仪技术测定血中Treg、Th17的含量,观察Treg/Th17比值变化.结果 (1)血Treg水平在治疗的第5天,正常对照组和模型组均高于酪酸梭菌组、蒙脱石散组、美沙拉秦组、美沙拉秦联合蒙脱石散组,差异有统计学意义(P<0.05);而在治疗的第12天,正常对照组和模型组明显低于酪酸梭菌组、蒙脱石散组、美沙拉秦组、美沙拉秦联合蒙脱石散组,尤其低于酪酸梭菌组,除美沙拉秦组外,差异有统计学意义(P<0.01);治疗的第12天和第5天相比,除正常组和模型组外,Treg值均有显著升高(P<0.01).(2)血Th17水平在治疗第5天和第12天时,正常对照组均明显低于模型组、酪酸梭菌组、蒙脱石散组、美沙拉秦组和美沙拉秦联合蒙脱石散组,尤其低于模型组,差异有统计学意义(P<0.05);治疗的第12天和第5天相比,Th17值除蒙脱石散组和酪酸梭菌组外,均明显降低,差异有统计学意义(P<0.05).(3)在治疗的第5天和第12天,正常组的Treg/Th17比值较其余各组高,尤其高于模型组(P<0.05).但在治疗的第12天,酪酸梭菌组、蒙脱石散组、美沙拉秦组、美沙拉秦联合蒙脱石散组的Treg/Th17比值已逐渐接近正常组,其中美沙拉秦联合蒙脱石散组升高最为明显(P<0.05),而模型组仍低;治疗的第12天和第5天相比,Treg/Th17比值均升高(P<0.05).结论 (1)Treg/Th17免疫失衡可能在UC发病中发挥重要作用;(2)酪酸梭菌、蒙脱石散、美沙拉秦能有效抑制或减低模型大鼠Th17的水平,提高血Treg细胞的水平,从而阻断炎症反应,缩短炎症的持续时间,其炎症控制与用药的时间也有关.

Objective To study the impa cts of clostridium, montmorillonite powder and mesalazine on Regulative T Cells（Treg） / Auxilliary T-17 Cells（Th17） immune balance in rats with ulcerative colitis（UC）induced by 2,4,6 TNBS/ethanol. Methods 48 rats were divided randomly into 6 groups（8 rats per group）including the model, clostridium, montmorillonite, mesalazine, mesalazine plus montmorillonite and normal group. TNBS/ethanol was used to build a model of rats with UC. The quantity of normal saline, clostridium,montmorillonite powder and mesalazine were converted by surface area of rat and given the rats by gastro-tube daily.4rats were executed on the 5th day after the molded and treatment, Other rats were executed on the 12 th day after the molded and treatment,and the blood samples were collected to measure the contents of Treg and Th17 by a flow cytometer and observe the Treg/Th17 ratios.Results（1） The levels of Treg in normal group and the model group were markedly higher than in clostridium,montmorillonite,mesalazine and mesalazine plus montmorillonite group on the fifth day, differences are statistically.（P〈0.05） However, this result reversed dramaticly on the 12 th day, which were more lower, especially in clostridium group.Significant difference appeared except the mesalazine group（P〈0.01）;Statistically comparing the 12 th day with the 5th day in the treatment, Treg values all rised tremendously, excluded the normal group and the model group.（2） The level of Th17 in normal group was obviously more lower than in other 5 groups on the 5th and 12 th day, apparently more lower than in the model group.The differences have statistic significance（P〈0.05）;Statistically comparing the 12 th day with the 5th day in the treatment,the Th17 value decreased apparently in the other 4 groups excluded the montmorillonite and clostridium group,differences were statistically significant（P〈0.05）.（3） The ratio of Treg/Th17 in normal group was obviously more higher than other 5 groups on the 5th and 12 th day, apparently higher than in the model group.（P 〈0.05） But on the 12 th day, the ratios of Treg/Th17 in clostridium, montmorillonite, mesalazine and mesalazine plus montmorillonite group were gradually close to normal group, especially in mesalazine plus montmorillonite group,（P〈0.05） while the model group still keeps lower level,Statistically comparing the 12 th day with the fifth day in the treatment, all the ratio of Treg/Th17 rised（P 〈0.05）.Conclusions（1） The immune imbalance of Treg/Th17 might play a important role in UC pathogenesis.（ 2） Clostridium,montmorillonite,and mesalazine could effectively restrain or reduce the level of Th17 and elevate the level of Treg in rats with UC model to prevent inflammatory reaction and shorten the duration of inflammation.