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目的:观察白介素-10(interleukin-10,IL-10)基因转化大肠杆菌(E.coli/mIL-10)对葡聚糖硫酸钠(DSS)诱导小鼠结肠炎的影响,并探讨其相关机制.方法:将小鼠IL-10基因序列转化至大肠杆菌(E.coli/mIL-10),阴性组为空质粒转化大肠杆菌(E.coli0).60只♀balb/c小鼠随机分成6组:正常组对照组,DSS组,DSS+E.coli/mIL-10组,DSS+E.coli0,E.coli/mIL-10组和E.coli0组.建立小鼠急性DSS结肠炎模型.自小鼠模型建立第1天开始,DSS+E.coli/mIL-10组和正常鼠+E.coli/mIL-10分别给予E.coli/mIL-10灌胃至实验结束,DSS+E.coli0组和E.coli0组分别给予E.coli0灌胃至实验结束(1×108cfu/天/只),正常对照组以及DSS组给予同等培养基灌胃至实验结束.每天观察各组疾病活动指数(DAI),并在实验结束后检测各组小鼠炎症肠段肿瘤坏死因子(TNF)、髓过氧化物酶(MPO)和核因子(NF)-κBP65的表达.结果:自实验第4天开始DSS+E.coli/mIL-10组小鼠DAI明显低于DSS组和DSS+E.coli0组.实验结束时DSS+E.coli/mIL-10组结肠组织中TNF(172.46±66.71pg/g组织)、MPO活性(2.35±0.39U/g组织)比DSS组和DSS+E.coli0组[(237.85±47.01)和(239.81±50.38)pg/g组织]、[(4.15±0.77)和(3.5±1.23)U/g组织]均明显降低;结肠组织NF-κB阳性表达减少.正常小鼠给予重组大肠杆菌灌胃后未出现结肠黏膜损伤.结论:利用经IL-10基因转化的大肠杆菌可以明显缓解DSS小鼠的结肠炎症损伤,减低MPO活性、抑制炎症肠段炎症细胞NF-κB的活化及炎性细胞因子的分泌.利用基因工程技术结合肠道共栖菌表达IL-10可以为IBD治疗提供一个新的方法.
Objective: To investigate the effect of interleukin-10 (IL-10) gene transformed E.coli / mIL-10 on colitis induced by dextran sodium sulfate (DSS) Methods: The mouse IL-10 gene sequence was transformed into E.coli / mIL-10, and the negative group was transformed into E.coli 0. The 60 Balb / c mice were randomly divided into 6 Group: DSS group, DSS + E.coli / mIL-10 group, DSS + E.coli0, E.coli / mIL-10 group and E.coli0 group.Acute DSS colitis model was established in mice. Starting from the first day after the establishment of the mouse model, E. coli / mIL-10 was administered to DSS + E. coli / mIL-10 group and normal mice + E. coli / mIL-10 orally until the end of the experiment. E. coli0 and E.coli0 groups were given intragastric administration of E.coli0 to the end of the experiment (1 × 108cfu / day / only), the normal control group and the DSS group were given the same medium until the end of the experiment.Each day observed disease activity index (DAI), and the expression of tumor necrosis factor (TNF), myeloperoxidase (MPO) and nuclear factor (NF) -κBP65 in inflammatory bowel of mice in each group were tested at the end of the experiment.Results: The DAI in starting DSS + E. coli / mIL-10 mice was significantly lower than that in DSS mice and DSS + E. coli0 At the end of experiment, TNF (172.46 ± 66.71pg / g tissue) and MPO activity (2.35 ± 0.39U / g tissue) in DSS + E.coli / mIL-10 group were significantly lower than those in DSS group and DSS + E.coli0 group [(237.85 ± 47.01) and (239.81 ± 50.38) pg / g tissues], [(4.15 ± 0.77) and (3.5 ± 1.23) U / g tissues] were significantly decreased, while the expression of NF- The colon mucosa injury was not observed in rats after administration of recombinant E.coli.Conclusion: The E.coli transformed with IL-10 gene can obviously relieve the colonic inflammatory injury, reduce the activity of MPO and inhibit the expression of NF-κB in inflammatory bowel inflammatory cells And the secretion of inflammatory cytokines.It is a new method for the treatment of IBD by using gene engineering technology combined with intestinal bacteria to express IL-10.