黄绿青霉素对肿瘤坏死因子α诱导的血管内皮细胞表达单核细胞趋化蛋白1和白细胞介素的影响

Effects of citreoviridin on the expression of MCP-1 and ILs induced by TNF-α in vein endothelial cells

目的观察黄绿青霉素(CIT)对血管内皮细胞表达单核细胞趋化蛋白1(MCP-1)、白细胞介素1β(IL-lβ)、IL-6和IL-8等细胞因子的影响,以及CIT上调肿瘤坏死因子α(TNF-α)诱导的内皮细胞表达MCP-1、IL-lβ、IL-6和IL-8的作用。方法体外原代培养人脐静脉内皮细胞(HUVECs),选择第5～6代进行试验,待细胞融合80%后随机分组,加入TNF-α(10μg/L)、CIT(2μmol/L)建立TNF-α组、CIT组、TNF-α+CIT联合处理组和空白对照组,处理时间24h。用酶联免疫吸附法(ELISA)检测细胞上清液IL-lβ、IL-6、IL-8及MCP-1的含量;免疫荧光染色法测定细胞核转录因子κB(NF-κB)的激活表达;RT-PCR检测各组MCP-1 mRNA表达。结果在TNF-α组和TNF-α+CIT组,细胞上清液IL-lβ、IL-6、IL-8、MCP-1浓度,细胞MCP-1 mRNA表达量,NF-κB P65蛋白表达量均高于空白对照组(P<0.05);且TNF-α+CIT组均高于TNF-α组(P<0.05)。结论 CIT可明显上调TNF-α诱导的内皮细胞MCP-1、IL-lβ、IL-6、IL-8表达和NF-κB的激活。

Objective To investigate the effects of citreoviridin（CIT） on the expression of MCP-1,IL-lβ,IL-6 and IL-8 induced by TNF-α in human umbilical vein endothelial cells（HUVECs）.Methods HUVECs isolated from the umbilical cord of neonates within 1 hour after birth（informed with consent form） were cultured in DMEM/F12 media.After 80% of HUVECs were confluent,the cells were divided into four groups and treated with CIT（2μmol/L） and/or TNF-α（10μg/L）.The levels of MCP-1,IL-lβ,IL-6 and IL-8 in the supernatant of cell culture media were measured by ELISA,the activation of NF-κB in HUVECs was detected by immunofluorescence staining,and the expression of MCP-1 mRNA was determinated by RT-PCR assay.Result The levels of MCP-1,IL-lβ,IL-6 and IL-8 in the supernatant and the expression of NF-κB P65 and MCP-1 mRNA of HUVECs were higher in TNF-α group and TNF-α＋CIT group than those in the control group（P 0.05）,and those of TNF-α＋CIT group was higher than TNF-α group（P 0.05）.Conclusion The expression of NF-κB,MCP-1,IL-lβ,IL-6 and IL-8 in HUVECs up-regulated by TNF-α was promoted by CIT.