基础医学与临床 ›› 2022, Vol. 42 ›› Issue (11): 1716-1720.doi: 10.16352/j.issn.1001-6325.2022.11.1716

• 研究论文 • 上一篇    下一篇

IL-35对可溶性CD40配体诱导血管内皮细胞损伤的影响

李明1, 孙海玲1, 刘娟利1, 郭巍巍2*   

  1. 南京医科大学康达学院附属滨海县人民医院 1.检验科; 2.介入与血管外科, 江苏 盐城 224500
  • 收稿日期:2021-11-02 修回日期:2022-04-07 出版日期:2022-11-05 发布日期:2022-11-01
  • 通讯作者: * guodapeng007@163.com
  • 基金资助:
    盐城市医学科技发展计划项目(YK2020080)

Effect of IL-35 on soluble CD40 ligand-induced injury of vascular endothelial cells

LI Ming1, SUN Hai-ling1, LIU Juan-li1, GUO Wei-wei2*   

  1. 1. Department of Laboratory Medicine; 2. Department of Interventional and Vascular Surgery, Binhai County People’s Hospital Affitiated to Kangda College of Nanjing Medical University, Yancheng 224500, China
  • Received:2021-11-02 Revised:2022-04-07 Online:2022-11-05 Published:2022-11-01
  • Contact: * guodapeng007@163.com

摘要: 目的 探索深静脉血栓(DVT)患者外周血可溶性CD40配体(sCD40L)和白细胞介素(IL)-35浓度,并研究IL-35对sCD40L诱导血管内皮细胞损伤的保护作用。方法 收集深静脉血栓急性期患者30例(DVT组)、健康对照者30名(HC组)外周静脉血3 mL。ELISA检测外周血清IL-35、sCD40L、IL-1β、IL-18的水平;体外培养人脐静脉血管内皮细胞(HUVECs)并分为3组,对照组(磷酸盐缓冲液)、sCD40L组(25 μg/mL sCD40L)、IL-35组(20 ng/mL IL-35和25 μg/mL sCD40L);CCK8法检测HUVECs的活力;ELISA检测细胞培养上清IL-1β、IL-18的水平;Western blot检测细胞GSDMD-N、cleaved caspase-1蛋白的表达。结果 DVT患者外周血IL-35水平显著低于对照者,sCD40L、IL-1β、IL-18的水平显著高于对照者(P<0.05)。DVT患者外周血IL-35与sCD40L呈负相关。体外实验表明:IL-35能抑制sCD40L诱导内皮细胞活力的损伤,降低sCD40L组细胞培养上清IL-1β、IL-18的水平,以及细胞GSDMD-N、cleaved caspase-1蛋白的表达。结论 DVT患者外周血IL-35水平降低,sCD40L水平升高。IL-35对sCD40L诱导血管内皮细胞损伤具有保护作用,可能影响血栓形成。

关键词: 白细胞介素-35, 可溶性CD40配体, 人脐静脉血管内皮细胞(HUVECs)

Abstract: Objective To investigate the concentration of soluble CD40 ligand (sCD40L) and interleukin (IL)-35 in peripheral blood of patients with deep vein thrombosis (DVT), and to study the protective effect of IL-35 on sCD40L-induced vascular endothelial cell injury. Methods Peripheral venous blood 3 mL was collected respectively from 30 patients with acute deep vein thrombosis patients (DVT group) and 30 healthy controls (HC group). Enzyme-linked immunosorbent assay (ELISA) was used to detect the level of serum IL-35, sCD40L, IL-1β, and IL-18. Human umbilical vein endothelial cells (HUVECs) were cultured and divided into three groups: control group (phosphate buffer), sCD40L group (25 μg/mL sCD40L), and IL-35 group (20 ng/mL IL-35 and 25 μg/mL sCD40L). CCK8 kit was used to detect the viability of HUVECs and ELISA was used to detect the level of IL-1β and IL-18 in the cell culture supernatant. The protein expression of GSDMD-N and cleaved caspase-1 in HUVECs was detected by Western blot. Results The level of IL-35 in peripheral blood of DVT patients was signifi- cantly lower than that in HC group, while the level of sCD40L, IL-1β, and IL-18 was significantly higher(P<0.05). Furthermore, there was a negative correlation between IL-35 and sCD40L in DVT patients. In vitro, IL-35 inhibited the damage of cells viability induced by sCD40L, and reduced the concentration of IL-1β and IL-18 in culture supernatant from sCD40L group. The protein expression of GSDMD-N and cleaved caspase-1 in cells was decreased too. Conclusions The level of IL-35 in peripheral blood of DVT patients is decreased, while the level of sCD40L is increased. IL-35 has a protective effect on sCD40L-induced vascular endothelial cell injury, which may have some impacts on the pathogenesis of thrombosis.

Key words: interleukin-35, soluble CD40 ligand, human umbilical vein endothelial cells(HUVECs)

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