IL-35对可溶性CD40配体诱导血管内皮细胞损伤的影响

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

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(25μg/mL sCD40L)、IL-35组(20 ng/mL IL-35和25μg/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。

Objective To investigate the concentration of soluble CD40 ligand(sCD40 L)and interleukin(IL)-35 in peripheral blood of patients with deep vein thrombosis(DVT),and to study the protective effect of IL-35 on sCD40 L-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,sCD40 L,IL-1β,and IL-18.Human umbilical vein endothelial cells(HUVECs)were cultured and divided into three groups:control group(phosphate buffer),sCD40 L group(25μg/mL sCD40 L),and IL-35 group(20 ng/mL IL-35 and 25μg/mL sCD40 L).CCK8 kit was used to detect the viability of HUVECs and ELISA was used to detect the level of IL-1βand 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 sCD40 L,IL-1β,and IL-18 was significantly higher(P<0.05).Furthermore,there was a negative correlation between IL-35 and sCD40 L in DVT patients.In vitro,IL-35 inhibited the damage of cells viability induced by sCD40 L,and reduced the concentration of IL-1βand IL-18 in culture supernatant from sCD40 L 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 sCD40 L is increased.IL-35 has a protective effect on sCD40 L-induced vascular endothelial cell injury,which may have some impacts on the pathogenesis of thrombosis.