百草枯中毒后NF-κB及其下游产物变化的研究

Expression of NF-κB and Its Downstream Products in Human Umbilical Vascular Endothelial Cells Treated with Paraquate

目的探讨百草枯处理后细胞内NF-κB的激活以及下游因子IL-6、IL-8的表达情况。方法体外培养人脐静脉血管内皮细胞(HUVEC)建立百草枯处理模型。在半数死亡浓度的百草枯作用8 h、24 h、48 h后,利用免疫细胞化学方法检测细胞NF-κB P65蛋白活化及定位,利用RT-PCR扩增IL-6、IL-8,半定量分析其mRNA表达水平变化。结果免疫组化显示正常细胞NF-κB P65定位于细胞胞浆,百草枯作用后开始出现细胞核转位,并随作用时间延长核转位数目明显增加;RT-PCR结果显示百草枯作用后NF-κB的下游产物IL-6、IL-8 mRNA表达水平均明显上调,并可持续到48 h。结论百草枯处理后早期即可能出现NF-κB的激活,进而诱导了下游的重要炎性介质的合成,促发和推进炎症反应,说明百草枯中毒后早期即有炎症反应参与了组织的损伤。

Objective To investigate the expression of NF-κB and its downstream products in the human umbilical vascular endothelial cells （HUVEC） treated with paraquate.Methods The HUVEC were cultured in vitro.The activation and location of NF-κB P65 protein were detected by immunocytochemical method at 8 h,24 h and 48 h after treatment with paraquat.The mRNA expression of IL-6 and IL-8 were detected by RT-PCR.Results Immunocytochemical analyses revealed that the activity subunit of NFκB P65 was located in cytoplasm of normal HUVEC.When the cells were treated with paraquat,positive staining in nuclear was observed.The sum of positive nuclear got more as the paraquat treated time increased.The RT-PCR verified the mRNA expression of IL-6 and IL-8 was up-regulated until 48 h after treatment with paraquat.Conlusion NF-κB could be activated at the early phase of paraquat poisoning.The activated NF-κB will induce the synthesis of inflammatory factors such as IL-6,IL-8 and then induce the inflammatory reaction.The inflammatory reaction have participated the tissue damage at the early phase of paraquat poisioning.