过敏性紫癜患儿外周血VDB...h17_Treg相关性研究_陈岑.pdf

过敏性紫癜患儿外周血 VDBP VD 表达水平与 Th17/Treg 相关性研究陈岑黄娜周玉超凌艳萍作者单位:210008江苏南京南京医科大学附属儿童医院风湿免疫科通信作者:凌艳萍，cc198010126 com 摘要 目的探讨维生素 D 结合蛋白(VDBP)、维生素 D 受体(VD)在过敏性紫癜(HSP)患儿外周血中的水平变化及与Th17/Treg 平衡的关系。方法选取 2019 年 12 月至 2020 年 12 月南京医科大学附属儿童医院就诊并确诊的 55 例 HSP 患儿为 HSP组，并以同期 50 例健康体检儿童为对照组。采用定量反转录 聚合酶链反应(qT PC)法检测外周血中 VD mNA 水平，酶联免疫法检测 VDBP 水平，蛋白印迹法检测 VD 蛋白的表达;流式细胞术检测 Th17 和 Treg 细胞比例;Pearson 法分析 VDBP 与 VD表达的相关性以及二者与 Th17/Treg 的相关性;受试者工作特征曲线(OC)评价血清 VDBP 和 VD 水平对 HSP 发生的诊断价值。结果与对照组比较，HSP 组患儿外周血中 VDBP 表达水平明显下调，VD 表达水平上调，差异均有统计学意义(P 0 05);HSP 组患儿 Th17、Th17/Treg 均高于对照组，Treg 均低于对照组(P 0 05);VDBP 与 Treg 呈正相关(r=0 358，P 0 05)，与 Th17、Th17/Treg 呈负相关(r=0 436、0 606，P 0 05);而 VD 与 Th17、Th17/Treg 均呈正相关(r=0 433、0 696，P 0 05)，与 Treg 呈负相关(r=0 385，P 0 05);HSP 患儿外周血中 VDBP 和 VD 的呈负相关(r=0 316，P 0 05);OC 结果显示，VD、VDBP 水平预测过敏性紫癜患儿的曲线下面积(AUC)分别为 0 732、0 933，对应的敏感度分别为 67 27%、94 55%，特异度分别为80 00%、78 00%，二者联合预测过敏性紫癜患儿的 AUC 为 0 946，灵敏感为 85 45%，特异度为 90 00%。结论VDBP 在 HSP 患儿外周血中明显低表达，VD 明显高表达，二者表达与 Th17/Treg 密切相关，联合检测 VD、VDBP 有助于诊断 HSP 的发生。关键词 过敏性紫癜;维生素 D 结合蛋白;维生素 D 受体;Th17/Tregdoi:10.3969/j.issn.1000 0399.2023.01.015Correlation between expression levels of VDBP and VD in peripheral blood and Th17/Treg in children with henoch schonleinpurpuraCHEN Cen，HUANG Na，ZHOU Yuchao，LING YanpingDepartment of heumatology and Lmmunology，Children s Hospital of Nanjing Medical University，Nanjing 210008，ChinaCorresponding author:LING Yanping，cc198010126 comAbstract ObjectiveTo explore the changes in the levels of vitamin D binding protein(VDBP)and vitamin D receptor(VD)inthe peripheral blood of children with Henoch Schonlein purpura(HSP)and their relationship with the balance of Th17/Treg MethodsAtotal of 55 children diagnosed with HSP in Children s Hospital Affiliated to Nanjing Medical University from December 2019 to December2020 were selected as the HSP group，and 50 healthy children in the same period were selected as the control group Quantitative reversetranscription polymerase chain reaction(QT PC)was used to detect the level of VD mNA in peripheral blood，and enzyme linkedimmunoassay was used to detect the level of VDBP Western blotting was applied to detect the expression of VD protein Flow cytometry wasused to detect the ratios of Th17 and Treg cells Pearson method was used to analyze the correlation between VDBP and VD expression andthe correlation between the two and Th17/Treg eceiver operating characteristic curve(OC)was used to evaluate the diagnostic value ofserum VDBP and VD levels for the occurrence of HSP esultsCompared with the control group，the expression level of VDBP in the pe-ripheral blood of children in the HSP group was significantly down regulated，the expression level of VD was up regulated，and thedifference was statistically significant(P 0 05)Th17 and Th17/Treg of children in HSP group were higher than those in control group，andTreg was lower than that in control group VDBP was positively correlated with Treg(r=0 358，P 0 05)，and negatively correlated withTh17 and Th17/Treg(r=0436，0606，P 005)VD was positively correlated with Th17 and Th17/Treg(r=0 433，0696，P 005)，and negatively correlated with Treg(r=0385，P 005)There was a significant negative correlation between VDBP and VD inperipheral blood of children with HSP(r=0316，P 005)OC results showed that the area under the curve(AUC)of VD and VDBPlevels predicting children with Henoch Schonlein purpura was 0732 and 0933，respectively，and the corresponding sensitivity was 6727%and 9455%，and the specificity was 8000%and 7800%，respectively The AUC of the combination of the two predicting children with He-noch Schonlein purpura was 0946，the sensitivity was 8545%，and the specificity was 9000%ConclusionsVDBP expression is signifi-47安徽医学第 44 卷第 1 期Anhui Medical Journal2023 年 1 月cantly low in peripheral blood of children with HSP，and VD expression is significantly high，which is closely related to Th17/Treg The com-bined detection of VD and VDBP contributes to the diagnosis of the occurrence of HSP，which has important clinical value Key words Henoch Schonlein purpura;Vitamin D binding protein;Vitamin D receptor;Th17/Treg过敏性紫癜(Henoch Schonlein purpura，HSP)是一种主要累及全身小血管的过敏性疾病，与自身免疫有关1。该病多发于儿童，临床表现为皮肤紫癜，伴有蛋白尿、腹痛、血尿等2 3，可在约四周内自行消退。但如果不能及时治疗，可能出现更严重的疾病，如持续性紫癜或关节炎，肠道受累导致内陷或大出血，以及进行性肾损伤4。因此对 HSP 进行早期诊断、加强防治，对临床工作具有重要意义。维生素 D 是一种类固醇激素，主要生理作用是维持钙稳态5。研究6 报道，维生素 D 与过敏性疾病关系紧密。维生素 D 受体(Vitamin D receptor，VD)属于核受体超家族，可以与维生素 D 的活性形式 1，25 二羟基维生素 D3 1，25(OH)2D3)结合，发挥生物效应7。据报道 VD 水平与系统性红斑狼疮、类风湿关节炎、I 型糖尿病等自身免疫性疾病有密切关系8。维生素 D 结合蛋白(vi-tamin D binding protein，VDBP)在代谢和生物途径中发挥不同作用9，但二者在 HSP 患儿中的作用尚不清楚。Th17 是一种新的促炎 T 辅助细胞，Treg 细胞是 T细胞的一个亚群，可以释放抗炎细胞因子 IL 10 和转化生长因子 TGF 1，发挥其抗炎特性。研究10 表明Th17/Treg 失衡与 HSP 的发病密切相关。本研究拟通过检测 VDBP、VD 在 HSP 患儿外周血中的水平，并探讨与 Th17/Treg 平衡的关系。现报道如下。1资料与方法1 1一般资料选取 2019 年 12 月至 2020 年 12 月南京医科大学附属儿童医院收治的 55 例 HSP 的患儿为研究对象，其中男性 23 例，女性 32 例，年龄 5 11岁，平均年龄(7 54 2 50)岁。选取50 例同期来本院体检的健康儿童为对照组，其中男性 30 例，女性 20例，年龄4 11 岁，平均年龄(7 64 2 70)岁。纳入标准:HSP 诊断参考诸福棠主编第 8 版实用儿科学11;首次发病;对照组均为体检结果健康的儿童;年龄12 岁。排除标准:合并有凝血功能障碍、自身免疫疾病;入院前进行激素、免疫抑制剂