重组人白细胞介素35(rhIL-35)通过抑制NF-κB通路减轻川崎病的冠状动脉内皮细胞损害  

Recombinant human interleukin 35(rhIL-35)alleviates the damage of coronary artery endothelial cells in Kawasaki disease by inhibiting NF-κB pathway

作　　者：田正 周忠 杨艳娟 李洁滢 胡琳 焦蓉 TIAN Zheng;ZHOU Zhong;YANG Yanjuan;LI Jieying;HU Lin;JIAO Rong(Department of Pediatrics,Graduate Training Base of Xiangyang First People's Hospital Affiliated to Hubei University of Medicine of Jinzhou Medical University,Xiangyang 441000,China;Department of Pediatrics,Xiangyang First People's Hospital,Hubei University of Medicine,Xiangyang 441000,China)

机构地区：[1]锦州医科大学湖北医药学院附属襄阳市第一人民医院研究生培养基地儿科,湖北襄阳441000 [2]湖北医药学院附属襄阳市第一人民医院儿科,湖北襄阳441000

出　　处：《细胞与分子免疫学杂志》2022年第10期880-885,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：湖北省襄阳市科技局计划项目(2020ZD34);湖北省儿科联盟科研项目(HPAMRP202108)。

摘　　要：目的探究重组人白细胞介素35(rhIL-35)对川崎病(KD)冠脉损害的保护作用及其作用机制。方法采用体外培养的人冠状动脉内皮细胞(HCAEC)建立KD血管模型,利用肿瘤坏死因子α(TNF-α)及KD患者血清刺激HCAEC以模拟KD的局部炎性病变。将细胞分为对照组、TNF-α及KD血清刺激组、(25、50)ng/mL rhIL-35处理组,CCK-8法检测细胞增殖活性,实时定量PCR检测HCAEC的IL-1β、IL-6、IL-17A、闭锁小带蛋白1(ZO-1)mRNA水平,ELISA检测KD患儿血浆IL-35水平及HCAEC上清液IL-1β、IL-6、IL-17A含量,Western blot法检测核因子κB p65(NF-κB p65)及ZO-1的表达。结果TNF-α及KD血清处理抑制HCAEC增殖,rhIL-35则明显逆转以上作用;rhIL-35预保护细胞后可明显下调IL-1β、IL-6及IL-17A的表达;与TNF-α及KD血清刺激组相比,rhIL-35预处理细胞后可明显增高ZO-1蛋白的表达,抑制NF-κB p65的高表达。结论rhIL-35能够通过抑制NF-κB通路减轻KD的冠状动脉内皮细胞损害。Objective To investigate the protective effect and mechanism of recombinant human interleukin 35(rhIL-35)on coronary artery injury in Kawasaki disease(KD).Methods Human coronary artery endothelial cells(HCAECs)were cultured in vitro to establish KD vascular model.Tumor necrosis factor α(TNF-α)and the serum of KD patients stimulated HCAECs were used to mimic the local inflammatory lesions of KD.The cells were divided into control group,TNF-α and KD serum stimulation group,(25,50)ng/mL rhIL-35 treatment group.Cell proliferation was detected by CCK-8 assay;mRNA levels of IL-1β、IL-6、IL-17A and zonula occludens-1(ZO-1)of HCAECs were detected by real-time quantitative PCR;IL-35 expression in plasma and IL-1β、IL-6 and IL-17A content in HCAEC supernatant were tested by ELISA;Western blot was performed to detect the expression of nuclear factor κB p65(NF-κB p65)and ZO-1.Results TNF-α and KD serum inhibited the proliferation of HCAECs,while rhIL-35 significantly reversed the above effects.RhIL-35 significantly down-regulated the expression of IL-1β,IL-6 and IL-17A after preconditioning HCAECs.Compared with TNF-α and KD serum stimulation group,rhIL-35 pretreated cells could significantly increase ZO-1 protein expression and inhibit NF-κB p65 expression.ConclusionsrhIL-35 can alleviate the damage of KD coronary artery endothelial cells by inhibiting NF-κB pathway.

关 键 词：川崎病(KD) 重组人白细胞介素35(rhIL-35) 冠状动脉内皮细胞(HCAEC) 炎性细胞因子 核因子κB(NF-κB) 

分 类 号：R725.9[医药卫生—儿科] R392.9[医药卫生—临床医学] R979.5[生物学—细胞生物学] Q291