白细胞介素-35对可溶性CD40配体诱导人脐静脉血管内皮细胞活化的作用

The effect of IL-35 on the activation of human umbilical vein endothelial cells induced by sCD40L

目的:探讨白细胞介素(IL)-35对可溶性CD40配体(sCD40L)诱导人脐静脉血管内皮细胞(HUVEC)活化的作用及IL-35和sCD40L在冠状动脉粥样硬化性心脏病(冠心病)发病过程中的作用。 方法:入选43例冠心病患者,其中不稳定型心绞痛20例(UA组),急性ST段抬高型心机梗死23例(STEMI组),20例健康体检者设为对照组,酶联免疫吸附试验(ELISA)检测外周血清IL-35和sCD40L水平,并分析两者的相关性。体外培养HUVEC,分为空白组、sCD40L组、IL-35+sCD40L组,ELISA法检测细胞培养上清E-选择素、可溶性细胞间黏附分子(sICAM-1)的水平;比色法检测HUVEC中丙二醛(MDA)水平及超氧化物歧化酶(SOD)活性;DCFH-DA荧光探针法检测HUVEC中氧自由基(ROS)活性。 结果:与对照组相比,UA组和STEMI组外周血清sCD40L水平显著升高,IL-35水平显著降低(P均<0.05),且两者呈负相关(r=-0.443,P<0.05)。体外培养HUVEC,与sCD40L组相比,IL-35+sCD40L组培养上清中E-选择素、sICAM-1水平以及HUVEC中MDA水平、ROS活性均显著降低,HUVEC中SOD活性显著升高(P均<0.05)。 结论:冠心病患者外周血清IL-35水平显著降低,sCD40L水平显著升高,IL-35对sCD40L活化HUVEC有抑制作用。

Objective:To explore the effect of interleukin (IL)- 35 on the activation of human umbilical vein endothelial cells(HUVEC)induced by soluble CD40 ligand(sCD40L)and the role of IL-35 and sCD40L in the pathogenesis of coronary heart disease(CHD). Methods:IL-35 and sCD40L serum levels were measured by ELISA from20 healthy subjects and 43 patients with CHD,including 20 cases with unstable angina pectoris(UA)and 23 cases with acute myocardial infarction (AMI). Pearson correlation was used to identify the relationship between IL-35 and sCD40L.HUVEC were cultured in vitro and divided into three groups:control group,sCD40L group and IL-35+sCD40L group.Concentration of E-selectin and soluble intracellular adhesion molecule-1(sICAM-1)in the cell culture supernatants was measured by ELISA.Colorimetric assay was used to evaluate malondialdehyde (MDA)and superoxide dismutase (SOD)activity levels in HUVEC.Intracellular reactive oxygen species(ROS)was determined by means of 2,7-diclorofluorescein diacetate (DCFH-DA)assay.Results:Compared with the control group,sCD40L levels significantly increased,while the level of IL-35 significantly decreased in the UA and AMI group(all P<0.05),and the level of IL-35 in the patients with CHD was negatively correlated with sCD40L (r=-0.443,P<0.05).In the IL-35+sCD40L group,the concentration of E-selectin and sICAM-1 in culture supernatants,and intracellular MDA and ROS activity levels decreased significantly,while the SOD activity increased,comparing with the sCD40L group(all P <0.05). Conclusions:IL-35 levels in peripheral blood of patients with CHD decreased,while the sCD40L levels increased.IL-35 was found to inhibit HUVEC activation induced by sCD40L.