维生素D3对脂多糖诱导流产小鼠外周血辅助性T细胞17/调节性T细胞的影响

Effect of vitamin D3 on helper T cell 17/regulatory T cell in peripheral blood of lipopolysaccharide induced abortion mice

目的探讨维生素D3对脂多糖诱导流产小鼠外周血辅助性T细胞17(Th17)/调节性T细胞(Treg)细胞的影响。方法将ICR孕小鼠随机分为正常组、模型组和实验组,每组10只;于妊娠第2天,实验组腹腔注射50 mg·L^-1维生素D30.2 mL,模型组和正常组注射等量0.9%NaCl,每日2次,至妊娠第7天模型组和实验组尾静脉注射脂多糖0.2 mL,正常组尾静脉注射等量生理盐水。以酶联免疫吸附法检测小鼠血清中炎性因子含量,以流式细胞术检测小鼠外周血中Th17和Treg百分比,观察各组小鼠胚胎孕育情况,以苏木精-伊红染色观察小鼠子宫组织病理学变化,以蛋白质印迹法检测各组小鼠子宫组织肿瘤坏死因子受体(TNFR)、B淋巴细胞瘤-xl(BCL-xl)蛋白表达情况。结果正常组、模型组和实验组小鼠流产率为10.00%(1只/10只),100.00%(10只/10只),50.00%(5只/10只),胚胎吸收率为8.62%(5只/58只),53.33%(32只/60只),35.48%(22只/62只),正常组、模型组和实验组小鼠白细胞介素-17A(IL-17A)/白细胞介素-10(IL-10)为0.19±0.05,4.28±0.13,0.69±0.25,Th17/Treg为0.23±0.08,1.67±0.21,0.58±0.13,子宫组织中TNFR蛋白表达量为0.23±0.04,1.02±0.33,0.56±0.19,BCL-xl蛋白蛋白表达量为0.96±0.11,0.52±0.13,0.79±0.15,差异均有统计学意义(均P<0.05)。结论维生素D3可改善脂多糖诱导流产小鼠妊娠结局,与调控Th17/Treg细胞平衡,调节子宫组织TNFR、BCL-xl蛋白表达有关。

Objective To investigate the effect of vitamin D3 on helper T cell 17(Th17)/regulatory T cell(Treg)in peripheral blood of lipopolysaccharide induced abortion mice.Methods ICR pregnant mice were randomly divided into normal group,model group and test group,10 mice in each group.On the second day of pregnancy,50 mg·L^-1 vitamin D30.2 mL was intraperitoneally injected into test group,0.9%NaCl was injected into model group and normal group twice a day,and lipopolysaccharide 0.2 mL was injected into tail vein of model group and test group on the seventh day of pregnancy,and normal saline 0.2 mL was injected into tail vein of normal group.Enzyme linked immunosorbent assay was used to detect the content of inflammatory factors in the serum of mice;flow cytometry was used to detect the percentage of Treg and Th17 cells in the peripheral blood of mice;embryogenesis of mice in each group was observed;hematoxylin eosin staining was used to observe the histopathological changes of mouse uterus;Western blotting was used to detect the expression of tumor necrosis factor receptor(TNFR)and B-lymphoma-xl(BCL-xl)protein of mouse uterus in each group.Results The abortion rates in normal group,model group and test group were 10.00%(1/10),100.00%(10/10)and 50.00%(5/10),embryo absorption rates were 8.62%(5/58),53.33%(32/60)and 35.48%(22/62),IL-17 A/IL-10 were 0.19±0.05,4.28±0.13,0.69±0.25,Th17/Treg were 0.23±0.08,1.67±0.21,0.58±0.13,respectively;The expression of TNFR protein were0.23±0.04,1.02±0.33 and 0.56±0.19,BCL-xl protein were 0.96±0.11,0.52±0.13 and 0.79±0.15,the difference were all statistically significant(all P<0.05).Conclusion Vitamin D3 can improve the pregnancy outcome of lipopolysaccharide induced abortion mice,which is related to the regulation of Th17/Treg cell balance and the regulation of TNFR and Bcl-xl protein in uterine tissue.