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目的探究不同剂量1,25-(OH)_2D_3对慢性溃疡性结肠炎小鼠炎症反应TLR4信号通路中相关因子的影响。方法 9w龄雌性C57BL/6小鼠50只,适应性喂养1w后,随机分为正常组、模型组、低、中、高剂量干预组。正常组小鼠在整个实验期间自由饮用蒸馏水,其余四组在第1~7 d、15~21 d自由饮3%的DSS溶液,其余时间自由饮用蒸馏水。在实验的第15~28日,低、中、高剂量干预组小鼠每天分别灌胃含10、50、100 ng 1,25-(OH)_2D_3 0.1ml的橄榄油,正常组和模型组小鼠则只灌胃0.1ml的橄榄油。于第29日处死全部小鼠。根据小鼠的一般情况,结合结肠大体形态评分以及结肠组织病理学评分对小鼠结肠严重程度进行评估。酶联免疫吸附法检测小鼠血清TNF-α、IL-10含量,免疫组化法检测小鼠结肠组织中VDR、TLR4、TRAF6及NF-κB1蛋白含量。结果与模型组小鼠相比,低、中、高剂量1,25-(OH)_2D_3干预组小鼠结肠大体形态和组织病理学评分均低于模型组,结肠长度均长于模型组。干预后,血清TNF-α含量降低,IL-10含量有所升高,结肠组织TLR4、NF-κB1、TRAF6表达减少,VDR蛋白表达增加。结论补充1,25-(OH)_2D_3可降低结肠组织TLR4、TRAF6、NF-κB1的表达,改善慢性溃疡性结肠炎,这可能与TLR4信号通路有关。
Objective To investigate the effects of 1,25- (OH) _2D_3 at different dosages on TLR4 signaling pathway in chronic ulcerative colitis mice. Methods 50 Wistar female C57BL / 6 mice were randomly divided into normal group, model group, low, middle and high dose intervention groups. The normal mice were allowed to drink distilled water for the entire duration of the experiment. The remaining four groups were allowed to drink 3% DSS solution on the 1st, 7th and 15th to 21st days, respectively, and the rest were free to drink distilled water. On the 15th to 28th day of the experiment, the mice in low, medium and high dose intervention groups were orally administered with olive oil (10,50,100 ng 1,25- (OH) _2D_3 0.1 ml daily, respectively. Rats only gavage 0.1ml of olive oil. All mice were sacrificed on the 29th day. According to the general situation of mice, colonic colon morphology score and colon histopathology score were used to evaluate the colonic severity of mice. The levels of TNF-α and IL-10 in serum were detected by enzyme-linked immunosorbent assay (ELISA). The contents of VDR, TLR4, TRAF6 and NF-κB1 in colonic tissues were detected by immunohistochemistry. Results Compared with the model group, the morphological and histopathological scores of colon in the low, medium and high dose 1,25- (OH) _2D_3 groups were lower than those in the untreated group, and the colon length were longer than that in the model group. After intervention, serum TNF-αlevel decreased and IL-10 content increased. The expression of TLR4, NF-κB1 and TRAF6 in colonic tissue decreased and the expression of VDR protein increased. Conclusion Supplementation of 1,25- (OH) _2D_3 can reduce the expression of TLR4, TRAF6 and NF-κB in colonic tissue and improve the chronic ulcerative colitis, which may be related to the TLR4 signaling pathway.