IL-8基因沉默后猪血管内皮细胞相关细胞因子表达的变化

Effects of IL-8 silencing on secretion of related cytokines in porcine vascular endothelial cell

为了探究内皮源IL-8对猪血管内皮细胞其他相关细胞因子表达的影响,解析IL-8在内皮细胞中作用,利用siRNA干扰技术对猪血管内皮细胞(VEC)中IL-8基因进行沉默,于干扰后不同时间收集细胞及上清,应用荧光定量PCR技术和ELISA检测VEC中各细胞因子的变化。结果显示,在mRNA水平上,促炎因子IL-6和促内皮生长因子(VEGF)在12h上调显著,且在4hVEGF显著上调;ICAM-1和VCAM-1在4h显著上调,其中ICAM-1在48h极显著下调,72h又显著上调;巨噬细胞集落刺激因子(M-CSF)几个时间段均极显著高于对照组。蛋白水平上,VEGF除48h外均显著高于对照组,M-CSF在4h后均显著低于对照组。以上数据表明,内皮源IL-8变化可能影响VEC增殖、迁移以及对造血干细胞分化。

In this study,it was aimed to dig out the effect of IL-8 on secretion of related cytokines in porcine vascular endothelial cell,and the small interfering RNA （siRNA） was applied to inhibit the expression of IL-8 in VEC.The related cytokines from VEC or its supernatants at different durations after RNAi,were assayed by real-time fluorescent quantitative PCR （Real time FQ-PCR） and ELISA respectively.The results showed the transcriptional changes of vascular endothelial growth factor （VEGF）,pro-inflammatory cytokines IL-6 in VECs were similar.Being Compared with the control group,IL-6 mRNA level from the interference one was significantly increased at 12 hours post interference （HPI） ; VEGF mRNA level was significantly increased at 4 and 12 HPI; ICAM-1 and VCAM-1 transcriptional levels were increased at 4 HPI,decreased at 48 HPI and increased at 72 HPI with significant difference for both at 4 HPI,ICAM at 48 HPI and 72 HPI.At the different durations,the expressions of M-CSF in the interference groups were all significantly higher than those in the control one.Meanwhile,VEGF expression was significantly increased except for 48 HPI and M-CSF expression was significantly decreased except for 4HPI.Collectively,all of the results indicated that silencing of IL-8 gene impact the secretion of VEGF,IL-6,hematopoietic growth factor M-CSF.It also implied that the change of IL-8 secretion in VEC could influence VEC＇s proliferation,migration and differentiation of hematopoietic stem cells.