目的：观察在同种异体角膜移植的小鼠模型中，1α，25-二羟维生素D3对Th17细胞及其相关细胞因子的影响。方法：C57BL／6小鼠作为受体，BALB／c小鼠作为供体，建立小鼠穿透性角膜移植术模型。术后隔日对角膜移植小鼠腹腔注射1.0μg 1α，25二羟维生素D3（维生素D3干预组），模型组和正常对照组腹腔注射等量大豆油。在裂隙灯活组织镜下观察角膜移植物的透明情况，评估排斥反应的发生情况，并取角膜移植物做病理学检测。采用实时荧光定量PCR检测脾脏中IL-17、RORγt和IFN-γ的表达水平，Western blotting分析外周血中RORγt及IL-17的蛋白含量，流式细胞术检测外周血IL-17和IFN-γ细胞因子的表达情况。结果：1α，25-二羟维生素D3显著地抑制了同种异体角膜移植排斥反应，同时减少了角膜移植物的炎性渗出。应用1α，25-二羟维生素D3治疗后，小鼠脾脏中IL-17、RORγt和IFN-γ的表达水平降低；外周血中IL-17、RORγt和IFN-γ的表达水平被下调。结论：1α，25-二羟维生素D3抑制小鼠同种异体角膜排斥反应的作用与IL-17及相关细胞因子RORγt细胞调节密切相关。%[ ABSTRACT] AIM:To evaluate the effects of 1α, 25-dihydroxyvitamin D3 on T helper cell 17 ( Th17 cells) and its related cytokines in a mouse model of corneal allograft transplantation.METHODS:C57BL/6 mice were transplanted with corneal grafts from BALB/c mice and treated intraperitoneally with 1.0μg 1α, 25-dihydroxyvitamin D3 or soybean oil every other day after operation.The transparency of the corneal grafts was evaluated for potential rejection signs by slit lamp biomicroscopy and histopathology.The expression levels of IL-17, RORγt and IFN-γin the spleen were measured by real-time PCR.Moreover, the protein expression of RORγt and IL-17 in the peripheral blood was analyzed by Western blotting. IL-17 and IFN-γin peripheral blood were measured by flow cytometry.RESULTS:1α, 25-dihydroxyvitamin D3 signifi-cantly inhibited the rejection of the corneal allograft and reduced the numbers of inflammatory infiltrates in the corneal graft. In the spleen, 1α, 25-dihydroxyvitamin D3 treatment reduced the expression levels of IL-17, RORγt and IFN-γ.In the pe-ripheral blood, 1α, 25-dihydroxyvitamin D3 treatment downregulated the expression levels of RORγt, IL-17 and IFN-γ. CONCLUSION:The effects of 1α, 25-dihydroxyvitamin D3 on suppressing corneal transplantation-induced allograft rejec-tion in mice are closely associated with its modulation on IL-17 and related cytokine RORγt.
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