【摘要】 目的：探究外源性维生素D（VitD）对肺炎链球菌诱导的慢性鼻-鼻窦炎（CRS）小鼠中调节性T细胞/辅助性T细胞17（Treg/Th17）功能失衡的作用。方法：将40只雄性C57BL/6小鼠按随机数字表法分为对照组（Control组）、模型组（Model组）、维生素D组（VitD组）和克拉霉素组（CLR组）,每组10只。除Control组外，其余各组小鼠均在鼻窦腔填充含肺炎链球菌菌液海绵以复制CRS模型，VitD组和CLR组小鼠分别以VitD₃溶液、克拉霉素溶液进行灌胃；21 d后，采用酶联免疫吸附试验检测小鼠血清白细胞介素（IL）6、IL-10、IL-17、IL-23及转化生长因子β（TGF-β）水平，苏木精-伊红染色和过碘酸雪夫染色观察小鼠鼻窦黏膜病理变化，实时荧光定量聚合酶链反应和蛋白质印迹法检测小鼠鼻窦黏膜内维甲酸相关孤儿受体γt（RORγt）和Forkhead家族转录因子3（Foxp3）的mRNA及蛋白表达水平，免疫组化染色检测小鼠鼻窦黏膜内IL-17和Foxp3阳性表达，流式细胞术分析外周血中CD4⁺IL-17A⁺和CD25⁺Foxp3⁺细胞比例分布。结果：与Control组比较，Model组小鼠血清中IL-6、IL-17及IL-23水平升高，IL-10、TGF-β水平降低（P<0.05）;鼻窦黏膜内发生明显病变，有大量炎症细胞浸润，杯状细胞数目增加（P<0.05）;RORγt mRNA和蛋白相对表达量上调，Foxp3 mRNA和蛋白相对表达量均下调（P<0.05）;IL-17阳性表达率升高，Foxp3阳性表达率降低（P<0.05）;外周血中CD4⁺IL-17A⁺细胞比例升高，CD25⁺Foxp3⁺细胞比例降低（P<0.05）,差异均有统计学意义。与Model组比较，VitD组和CLR组小鼠血清中IL-6、IL-17和IL-23水平降低，IL-10、TGF-β水平升高（P<0.05）;鼻窦黏膜内炎症细胞浸润减轻，杯状细胞数目减少（P<0.05）;RORγt mRNA和蛋白相对表达量均下调，Foxp3 mRNA和蛋白相对表达量均上调（P<0.05）;IL-17阳性表达率降低，Foxp3阳性表达率升高（P<0.05）;外周血中CD4⁺IL-17A⁺细胞比例降低，CD25⁺Foxp3⁺细胞比例升高（P<0.05）,差异均有统计学意义。结论：VitD能够改善CRS模型小鼠鼻窦黏膜的病理表现，减轻炎症细胞浸润，其机制可能与调控Treg/Th17细胞功能失衡相关。更多还原

【Abstract】 OBJECTIVE: To probe into the role of vitamin D（VitD） in improving chronic rhinosinusitis（CRS） model induced by Streptococcus pneumoniae by regulating the imbalance of T regulatory cells/T helper 17 cells（Treg/Th17） cell function. METHODS: Totally 40 male C57BL/6 mice were divided into the control group（Control）, model group（Model）, VitD group（VitD） and clarithromycin group（CLR） according to the random table method, with 10 mice in each group. Except for the control group, the other groups were filled with sponges containing S. pneumoniae in the sinus cavity to replicate the CRS model. The VitD group and CLR group were administered with VitD₃ solution and clarithromycin solution by gavage respectively. After 21 d, the levels of interleukin（IL）-6, IL-10, IL-17, IL-23 and transforming growth factor-β（TGF-β） in the serum of mice were detected by enzyme linked immunosorbent assay. Hematoxylin and Eeosin staining and periodic acid-schiff staining were used to observe the pathological changes of mucous membranes of the nasal sinuses of mice, real-time quantitative polymerase chain reaction and Western blotting were used to detect the mRNA and protein expression levels of retinoic acid-related orphan receptor γt（RORγt） and forkhead family transcription factor 3（Foxp3） in the mucous membranes of the nasal sinuses of mice, immunohistochemical staining was used to detect the positive expression of IL-17 and Foxp3 in the sinus mucosa of mice, and flow cytometry was used to analyze the distribution of CD4⁺IL-17A⁺ and CD25⁺Foxp3⁺ cells in peripheral blood. RESULTS: Compared with the control group, the serum levels of IL-6, IL-17 and IL-23 in the model group increased, while the levels of IL-10 and TGF-β decreased（P<0.05）; obvioued lesions occurred in the sinus mucosa, with a large number of inflammatory cells infiltrating, the number of goblet cells increased（P<0.05）; the relative expression of RORγt mRNA and protein was up-regulated, and the relative expression of Foxp3 mRNA and protein was down-regulated（P<0.05）; the positive expression rate of IL-17 increased, while the positive expression rate of Foxp3 decreased（P<0.05）, the proportion of CD4⁺IL-17A⁺cells in peripheral blood increased, and the proportion of CD25⁺Foxp3⁺ cells decreased（P<0.05）, with statistically significant differences. Compared with the model group, the serum levels of IL-6, IL-17 and IL-23 in the VitD group and CLR group decreased, while the levels of IL-10 and TGF-β increased（P<0.05）; the infiltration was alleviated, and the number of goblet cells decreased（P<0.05）; at the same time, the relative expressions of RORγt mRNA and protein were down-regulated, the relative expressions of Foxp3 mRNA and protein were up-regulated（P<0.05）, the positive expression rate of IL-17 decreased and the positive expression rate of Foxp3 increased（P<0.05）, the proportion of CD4⁺IL-17A⁺ cells in peripheral blood decreased and the proportion of CD25⁺Foxp3⁺ cells increased（P<0.05）, with statistically significant differences. CONCLUSIONS: VitD can improve the pathological manifestations of sinus mucosa in CRS model mice and reduce inflammatory cell infiltration, and its mechanism may be related to the regulation of imbalance of Treg/Th17 cell function.更多还原