川崎病高表达SEMA7A促进单核和中性粒细胞黏附与迁移  

High expression of SEMA7A in Kawasaki disease promotes adhesion and migration of monocytes and neutrophils

作　　者：黄君华 党心睿 方可 苏以萱 李一阳 张书婉 HUANG Junhua;DANG Xinrui;FANG Ke;SU Yixuan;LI Yiyang;ZHANG Shuwan(Basic Clinical Laboratory and Hemato-logy Laboratory Teaching and Research Section,School of Medical Technology,Xi'an Medical University,Xi'an 710021,China;School of Medical Technology,Xi'an Medical University;Clinical Laboratory,Xi'an Children's Hospital)

机构地区：[1]西安医学院医学技术学院临床检验基础与血液学检验教研室,西安710021 [2]西安医学院医学技术学院 [3]西安市儿童医院检验科

出　　处：《山西医科大学学报》2025年第1期43-48,共6页Journal of Shanxi Medical University

基　　金：陕西省自然科学基础研究计划项目(2024JC-YBQN-0869,2023-JC-YB-718);西安市科技计划项目(24YXYJ0064);西安市卫生健康委员会科研项目(2023qn12);陕西省大学生创新创业训练计划项目(S202311840077,S202311840070);西安医学院大学生创新创业训练计划项目(121523077,121523070)。

摘　　要：目的 探究川崎病(KD)条件下Semaphorin 7A(SEMA7A)对单核细胞(THP-1细胞系)和中性粒细胞(HL-60细胞系)黏附与跨内皮细胞迁移的作用。方法 将4周龄雄性C57BL/6小鼠随机分为对照组和模型组,每组6只;模型组小鼠腹腔注射干酪乳杆菌细胞壁提取物(LCWE)构建KD样血管炎小鼠模型,对照组腹腔注射等量磷酸盐缓冲液(PBS);注射后第14天采用实时定量PCR(RT-PCR)和免疫印迹(Western blot)检测心脏组织SEMA7A表达情况。体外培养人冠状动脉内皮细胞(HCAEC)并将细胞分为健康儿童(HC)血清刺激组和KD血清刺激组,培养24 h后采用RT-PCR和Western blot检测细胞SEMA7A表达水平。KD血清刺激HCAEC分为si-NC组和si-SEMA7A组,采用RT-PCR法检测敲减效率。将四组HCAEC分别接种于Transwell上室形成细胞单层,与THP-1和HL-60细胞分别共培养后,利用荧光标记和Transwell实验检测黏附和跨内皮迁移的细胞数量。结果 与对照组相比,模型组小鼠心脏组织SEMA7A的mRNA和蛋白表达均升高(P<0.01)。体外实验发现,KD血清刺激组SEMA7A的mRNA和蛋白表达均高于HC血清刺激组(P<0.01),si-SEMA7A组SEMA7A的mRNA表达低于siNC组(P<0.01)。KD血清刺激组HCAEC单层黏附的THP-1和HL-60细胞数量高于HC血清刺激组(P<0.05),si-SEMA7A组黏附的THP-1和HL-60细胞数量低于si-NC组(P<0.05)。Transwell迁移实验表明,KD血清刺激组THP-1和HL-60跨内皮细胞迁移的数量高于HC血清刺激组(P<0.01),si-SEMA7A组THP-1和HL-60跨内皮细胞迁移的数量较si-NC组明显下降(P<0.05)。结论 川崎病条件下高表达的SEMA7A能够促进单核细胞和中性粒细胞的内皮细胞黏附与和跨内皮细胞迁移。Objective To investigate the role of Semaphorin 7A(SEMA7A)in the adhesion and the transendothelial migration of monocytes(THP-1 cell line)and neutrophils(HL-60 cell line)in Kawasaki disease(KD).Methods For in vivo experiment,four-week-old male C57BL/6 mice were randomly divided into control group and model group,with 6 mice in each group.The mice in model group were intraperitoneally injected with lactobacillus casei cell wall extract(LCWE)to establish a KD-like vasculitis mouse model,while the mice in control group were injected with phosphate-buffered saline(PBS).At day 14 after injection,the expression of SEMA7A in cardiac tissue was detected by real-time quantitative PCR(RT-PCR)and Western blot.For in vitro experiment,human coronary artery endothelial cells(HCAECs)were divided into healthy children's serum(HC)stimulation group and KD serum stimulation group.After 24 h culture,the expression level of SEMA7A was detected by RT-PCR and Western blot.Moreover,HCAECs stimulated by KD serum were further divided into si-NC group and si-SEMA7A group,and the SEMA7A expression was detected by RT-PCR.Furthermore,the HCAECs from four groups were respectively seeded into the upper chamber of Transwell device to form a monolayer and co-cultured with THP-1 or HL-60,then the adhesion and the transendothelial migration numbers of THP-1 and HL-60 were detected by fluorescence labeling and Transwell experiment.Results Compared with control group,the mRNA and protein expressions of SEMA7A in cardiac tissue of model group were increased(P<0.01).The mRNA and protein expressions of SEMA7A in KD serum stimulation group were both higher than those in HC serum stimulation group(P<0.01).Additionally,the mRNA expression of SEMA7A in si-SEMA7A group was lower than that in si-NC group(P<0.01).Moreover,the numbers of THP-1 and HL-60 adhering to HCAEC monolayer in KD serum stimulation group were higher than those in HC serum stimulation group(P<0.05),and the numbers in si-SEMA7A group were lower than those in si-NC group(P<0.05).Furthermore,

关 键 词：信号素7A 川崎病 单核细胞 中性粒细胞 内皮细胞 细胞黏附 迁移 

分 类 号：R725[医药卫生—儿科]