川崎病中性粒细胞高表达CD100促进冠状动脉内皮细胞迁移和炎症反应  

The High Expression of CD100 in Neutrophils in Patients with KawasakiDisease Promotes the Migration and Inflammatory Response ofCoronary Artery Endothelial Cells

作　　者：黄君华 谢小娟[2] 赵传梅 张书婉 HUANG Junhua;XIE Xiaojuan;ZHAO Chuanmei;ZHANG Shuwan(Department of Medical Technology,Xi’an Medical University,Xi’an 710021,China;Shaanxi Center For Clinical Laboratory,Shaanxi Provincial People’s Hospital,Xi’an 710068,China;Clinical Laboratory,Xi’an Children’s Hospital,Xi’an 710003,China)

机构地区：[1]西安医学院医学技术学院,陕西西安710021 [2]陕西省人民医院省临检中心,陕西西安710068 [3]西安市儿童医院检验科,陕西西安710003

出　　处：《标记免疫分析与临床》2023年第11期1831-1835,共5页Labeled Immunoassays and Clinical Medicine

基　　金：陕西省自然科学基础研究计划项目(编号:2023-JC-YB-718);西安市卫生健康委员会科研项目(编号:2023qn12);西安市未央区科技计划项目(编号:202222);西安医学院科研能力提升计划项目(编号:2022NLTS107)。

摘　　要：目的探讨川崎病(KD)患者中性粒细胞条件培养基对冠状动脉内皮细胞(HCAECs)生物学功能的影响。方法以健康儿童(HC)外周血中性粒细胞为对照组,KD患儿外周血中性粒细胞为实验组,KD中性粒细胞通过siRNA沉默CD100为si-CD100组,无效siRNA转染为si-NC组,RT-PCR检测siRNA敲减效率及CD100 mRNA表达;体外培养各组中性粒细胞并收集条件培养基,ELISA法检测各组条件培养基中CD100的水平;Transwell实验检测各组中性粒细胞条件培养基对HCAECs迁移能力的影响;RT-PCR检测各组中性粒细胞条件培养基对HCAECs细胞因子IL-1β和IL-18表达水平的影响。结果RT-PCR显示KD中性粒细胞CD100 mRNA表达较HC组明显升高(t=11.92,P<0.05),验证si-CD100组CD100 mRNA的表达较si-NC组明显降低(t=13.40,P<0.05);ELISA实验结果显示,KD中性粒细胞条件培养基中CD100水平较HC组明显增高(t=12.01,P<0.05),si-CD100组较si-NC组明显降低(t=11.53,P<0.05);Transwell实验结果显示,与HC相比,经KD中性粒细胞条件培养基处理的HCAECs的迁移能力明显增加(t=12.63,P<0.05),而si-CD100组较si-NC组明显下降(t=4.09,P<0.05);RT-PCR实验结果显示,与HC相比,经KD中性粒细胞条件培养基处理的HCAECs中IL-1β和IL-18的mRNA表达明显升高(t=14.11和18.40,P<0.05),而si-CD100组较si-NC组明显减低(t=11.88和9.21,P<0.05)。结论川崎病患者中性粒细胞高表达并分泌CD100,CD100促进冠状动脉内皮细胞迁移和炎症反应。Objective To investigate effects of Kawasaki disease(KD)patients’neutrophil-conditioned medium on the biological functions of human coronary artery endothelial cells(HCAECs).Methods Neutrophils from peripheral blood of healthy children(HC)were used as the control group,while neutrophils from KD patients were used as the experimental group.KD neutrophils were transfected with siRNA to silence CD100(si-CD100 group),and ineffective siRNA transfection was used as the negative control(si-NC group).RT-PCR was performed to assess the knockdown efficiency of siRNA and CD100 mRNA expression.Neutrophils from different groups were cultured in vitro,and the conditioned medium was collected.The concentration of CD100 in each conditioned medium was measured using ELISA.Trans-well assays were conducted to evaluate the migration ability of HCAECs treated with neutrophil-conditioned medium from different groups.RT-PCR was performed to assess the expression levels of the cytokines IL-1βand IL-18 in HCAECs treated with neutrophil-conditioned medium.Results RT-PCR results showed a significantly increased CD100 mRNA expression level in KD neutrophils compared to the HC group(t=11.92,P<0.05),while the si-CD100 group exhibited a significantly decreased CD100 mRNA expression level compared to the si-NC group(t=13.40,P<0.05).ELISA results demonstrated that CD100 concentration in the conditioned medium from KD neutrophils was significantly higher than that in HC group(t=12.01,P<0.05),and si-CD100 group exhibited significantly lower CD100 concentration compared to si-NC group(t=11.53,P<0.05).Trans-well assay results revealed that the migration ability of HCAECs treated with conditioned medium from KD neutrophils was significantly increased compared to HC group(t=12.63,P<0.05),while si-CD100 group exhibited a significant decrease compared to si-NC group(t=4.09,P<0.05).RT-PCR results showed that the mRNA expression levels of IL-1βand IL-18 in HCAECs treated with conditioned medium from KD neutrophils were significantly elevated compared

关 键 词：CD100 川崎病 中性粒细胞 冠状动脉内皮细胞 儿童 

分 类 号：R725.4[医药卫生—儿科]