摘要
目的探讨益生菌VSL#3对2,4,6-三硝基苯磺酸(TNBS)诱导的溃疡性结肠炎大鼠调节性T细胞表达的影响及其作用机制。方法将SD大鼠随机分为5组:对照组、模型组、VSL#3组、Wortmannin组和VSL#3+Wortmannin组,对照组大鼠正常喂养,其余4组利用TNBS诱导建立溃疡性结肠炎大鼠模型,VSL#3组于模型建立后给予VSL#3灌胃,Wortmannin组腹腔注射Wortmannin溶液,VSL#3+Wortmannin组先以VSL#3灌胃再腹腔注射Wortmannin溶液。大鼠继续喂养2周,观察大鼠喂养期一般情况,计算疾病活动指数(DAI)。2周后处死大鼠,观察结肠组织学改变,检测结肠组织髓过氧化物酶(MPO)活性和磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)通路相关蛋白表达;同时检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-10含量和外周血调节性T细胞(Treg)比例。结果从第7天开始,VSL#3组、Wortmannin组DAI评分明显低于同时段模型组,VSL#3+Wortmannin组DAI评分较模型组降低更明显,差异有统计学意义(P<0.05)。VSL#3组和Wortmannin组MPO活性和TNF-α、IL-6水平明显低于模型组,IL-10水平明显高于模型组,VSL#3+Wortmannin组则更显著,各组差异有统计学意义(均P<0.05)。与模型组比较,VSL#3组和Wortmannin组CD4^+CD25^+Treg细胞比例和Treg/CD4^+T细胞比值明显升高;VSL#3+Wortmannin组升高得更明显,差异有统计学意义(均P<0.01)。此外,VSL#3组、Wortmannin组、VSL#3+Wortmannin组p-Akt、NF-κB p65蛋白表达明显低于模型组,差异有统计学意义(均P<0.01)。结论 VSL#3能减轻溃疡性结肠炎大鼠炎症反应和稳定Treg细胞/CD4^+T细胞比值,其作用机制可能与抑制PI3K/Akt信号通路有关。
Objective To examine the effects of VSL#3 probiotics on TNBS-induced colitis in rats and the underlying mechanism.Methods Sprague-Dawley rats were randomly divided into control group,model group,VSL#3 group,Wortmannin group and VSL#3+Wortmannin group.Rats in control group were dealt with routinely feeding.TNBS-induced colitis rat model was established in the other groups.In VSL#3 group,the rats were treated with VSL#3.The rats in Wortmannin group were administrated with Wortmannin.The rats in VSL#3+Wortmannin group were given VSL#3 combined with Wortmannin.After 2 weeks of induction,the disease activity index(DAI)was calculated,and then rats were euthanized and the colon was removed,the morphological change of rats’ colonic membrane was observed,the MPO activity and the expression of PI3 K/Akt pathway related protein was detected.Meanwhile,the levels of serum TNF-α,IL-6,IL-10 and Treg cells ratio were also examined.Results From the day 7,DAI score was significantly lower in the VSL#3 group and wortmannin group than in the model group,and DAI score decreased more significantly in the VSL#3+Wortmannin group(P<0.05).MPO activity and levels of TNF-α and IL-6 were significantly lower,IL-10 was significantly higher in the VSL#3 group and Wortmannin group than in the control group(all P<0.05).Compared to the control group,CD4^+CD25^+Treg cell ratio and Treg/CD4^+T cell ratio were significantly increased in the VSL#3 group and Wortmannin group,and VSL#3+Wortmannin group increased more obviously,the difference was statistically significant(all P<0.05).The protein expression levels of p-Akt and NF-κB p65 were significantly lower in the VSL#3,Wortmannin and VSL#3+Wortmannin groups than in the control group(all P<0.05).Conclusion VSL#3 can alleviate the inflammation and maintain the ratio of Treg/CD4^+T cells of rats with ulcerative colitis,which might be related with inhibition of PI3 K/Akt signaling pathway.
作者
罗燕
陈志涛
徐崇明
张曲
Luo Yan;Chen Zhitao;Xu Chongming(Department of Abdominal Radiotherapy,Hubei Cancer Hospital, Hubei Clinical Medical Research Center for Colorectal Cancer,Wuhan Colorectal Cancer Clinical Medical Research Center,Wuhan 430079;Department of Gastroenterology,The Central Hospital of Wuhan A.ffiliated to Totigji Medical College,Huazhong University of Science and Technology,Wuhan 430014,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2019年第4期411-416,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金青年基金资助项目(No.81400578)
