重组可溶性人CD40L诱导人脐静脉内皮细胞损伤

目的:探讨重组可溶性人CD40L(rsh CD40L)对人脐静脉内皮细胞(HUVECs)的损伤作用及其在动脉粥样硬化中的作用。方法:应用rsh CD40L刺激人脐静脉内皮细胞12 h;MTS法观察HUVECs的生存活性,ELISA法测内皮细胞E-选择素(E-selectin)、细胞间黏附分子(ICAM)-1、组织因子(TF)、组织因子途径抑制物(TFPI)表达的变化,比色法测脂质过氧化物丙二醛(MDA)含量及超氧化物歧化酶(SOD)活力。结果:与正常组比较,不同浓度的rsh CD40L(0.5、1、2、3 mg/L)对内皮细胞的生存活性无明显影响;0.5 mg/L rsh CD40L即可增加内皮细胞E-selectin、s ICAM-1、TF、TFPI的分泌,差异有统计学意义(P0.01),同时增加内皮细胞MDA的含量、降低SOD活性(P0.05)。结论:0.5~3 mg/L rsh CD40L对内皮细胞生存活性无明显影响,但已经引起内皮细胞功能障碍,增加内皮细胞炎症和外源性凝血反应,诱导内皮细胞脂质过氧化物损,使其抗氧化能力下降。

Injury effect of recombinant soluble human CD40 ligand on human umbilical vein endothelial cells

AIM: To investigate the damage in human umbilical vein endothelial cells (HUVECs) induced by recombinant soluble human CD40 ligand (rshCD40L). METHODS: The cultured HUVECs were treated with rshCD40L for 12 h. The survival activity of the HUVECs was observed by MTS assay. The expression of E-selectin, intercellular adhesion molecule (ICAM)-1, tissue factor (TF) and tissue factor pathway inhibitor (TFPI) was measured by ELISA. The activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) were detected by the methods of thibabituric acid (TBA). RESULTS: Compared with normal group, different concentrations of rshCD40L (0.5, 1, 2, 3 mg/L) had no obvious effect on the survival activity of the HUVECs (P>0.05). rshCD40L at concentration of 0.5 mg/L promoted the secretion of E-selectin, sICAM-1, TF and TFPI in the HUVECs (P<0.01). rshCD40L at concentration of 0.5 mg/L also increased MDA content and reduced the activity of SOD in the HUVECs (P<0.05). CONCLUSION: 0.5~3mg/L rshCD40L has no obvious effect on endothelial cell survival, but already causes endothelial dysfunction by increasing endothelial inflammation and exogenous coagulation reaction, inducing lipid peroxides injury and reducing antioxidant capacity.