维生素D、辅助性T淋巴细胞17和调节性T淋巴细胞及相关细胞因子在多发性硬化青少年患者中的变化及其意义

Changes and their significance of vitamin D,T helper cell 17 and regulatory T cells and associated cytokines in adolescents with multiple sclerosis

目的探讨外周血维生素D、辅助性T淋巴细胞17(Th17细胞)和调节性T淋巴细胞(Treg细胞)表达率以及血清中肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)水平在多发性硬化(MS)青少年患者发病机制中的作用。方法选取2010年1月至2020年12月新乡医学院第一附属医院儿科确诊为MS的38例青少年患者作为MS组,同期的38名年龄及体质量指数相匹配的健康者作为健康对照组。采用液相色谱仪检测MS青少年患者(MS组)与健康对照者(健康对照组)外周血25-羟基维生素D3[25-(OH)D3]水平,流式细胞术检测其Th17细胞和Treg细胞的表达率,免疫荧光法检测血清中TNF-α、MCP-1水平。采用SPSS 22.0统计学软件分析MS组与健康对照组上述指标之间的差异及各指标间的相关性。结果MS组与健康对照组相比,外周血25-(OH)D3水平差异无统计学意义(P>0.05);Th17细胞表达率明显高于健康对照组[(3.02±0.20)%比(1.99±0.16)%,t=12.03,P<0.05];外周血Treg细胞表达率明显低于健康对照组[(4.63±0.77)%比(5.10±0.90)%,t=14.65,P<0.05];Th17/Treg细胞的比值明显高于健康对照组(0.25±0.07比0.17±0.05,t=16.89,P<0.05);外周血TNF-α、MCP-1水平均高于健康对照组[(26.13±5.98)ng/L比(24.45±3.01)ng/L;(122.26±37.71)ng/L比(87.95±17.66)ng/L,t=1.986、47.650,均P<0.05]。MS组中,外周血25-(OH)D3与其他检测指标均无相关性;疾病复发或进展期患儿TNF-α、MCP-1水平均高于疾病缓解稳定期患儿[(17.49±3.94)ng/L比(14.45±3.81)ng/L;(90.42±23.06)ng/L比(77.55±20.56)ng/L,t=1.990、2.472,均P<0.05];Th17细胞的表达率与其外周血中TNF-α、MCP-1水平均呈正相关(r=0.478、0.442,均P<0.05);Treg细胞的表达率与其外周血中的TNF-α、MCP-1水平均呈负相关(r=-0.318、-0.356,均P<0.05)。结论Th17细胞和Treg细胞可能参与MS青少年患者发病的免疫病理机制;相关细胞因子的变化可能参与Th17细胞和Treg细胞变化的调控及炎症反应;Th17细胞、Treg细胞及相关细胞因子变化可能在MS的发生、发展、复发进展中起重要免疫病理机制作用。

Objective To investigate the function of the expression rate of vitamin D,T helper cell 17(Th17)and regulatory T cells(Tregs)and the serum level of tumor necrosis factor-α(TNF-α)and monocyte chemotactic protein-1(MCP-1)in the pathogenesis of adolescents with multiple sclerosis(MS).Methods Thirty-eight adolescent patients diagnosed with MS in the Department of Pediatrics of the First Affiliated Hospital of Xinxiang Medical University from January 2010 to December 2020 were selected as the MS group,and 38 healthy adolescents matched with age and body mass index were selected as healthy control group.The level of 25-hydroxyvitamin D3[25-(OH)D3]was measured by liquid chromatograph;the expression rates of Th17 and Tregs were measured by flow cytometry;the serum levels of TNF-αand MCP-1 were measured by immunofluorescence.SPSS 22.0 statistical software was adopted to analyze the differences between the above indicators and their correlation in the MS group and the healthy control group.Results There was no difference in the level of 25-(OH)D3 in peripheral blood between the MS group and the healthy control group(P>0.05).As for the MS group,the expression rate of Th17 was significantly higher than that in the healthy control group[(3.02±0.20)%vs.(1.99±0.16)%,t=12.03,P<0.05];the expression rate of Tregs in peripheral blood was significantly lower than that in the healthy control group[(4.63±0.77)%vs.(5.10±0.90)%,t=14.65,P<0.05];the ratio of Th17 to Tregs was significantly higher than that in the healthy control group(0.25±0.07 vs.0.17±0.05,t=16.89,P<0.05);the levels of TNF-αand MCP-1 in peripheral blood were higher than those in the healthy control group[(26.13±5.98)ng/L vs.(24.45±3.01)ng/L;(122.26±37.71)ng/L vs.(87.95±17.66)ng/L,t=1.986,47.650,all P<0.05].In the MS group,25-(OH)D3 in peripheral blood was not correlated with other test indicators;the levels of TNF-αand MCP-1 were higher in patients with relapsing or progressive disease than in patients with stable disease in remission[(17.49±3.94)ng/L vs.(14.45±3.81)ng/L;(90.42±23.06)ng/L vs.(77.55±20.56)ng/L,t=1.990,2.472,all P<0.05];the expression rate of Th17 cells was positively correlated with the level of TNF-αand MCP-1 in peripheral blood(r=0.478,0.442,all P<0.05);the expression rate of Tregs was negatively correlated with the level of TNF-αand MCP-1 in peripheral blood(r=-0.318,-0.356,all P<0.05).Conclusions Th17 and Tregs may be involved in the immunopathological mechanisms in the pathogenesis of adolescents with MS;changes in associated cytokines may be involved in the regulation of Th17 and Tregs changes and the inflammatory response;Th17 and Tregs and associated cytokine changes may play an important role in the occurrence,progression,and relapse progression of MS as an immunopathological mechanism.