摘要
目的探讨壳寡糖对血管内皮细胞损伤的保护机制。方法以EA.hy926血管内皮细胞为研究对象,通过TNF-α刺激,建立高表达IL-6、IL-8的内皮细胞损伤模型。以RT-PCR和ELISA方法分别从mRNA和蛋白水平检测壳寡糖对IL-6、IL-8表达的影响。采用Western blot方法检测壳寡糖对p38MAPK信号通路及糖基转移酶(OGT)表达的影响。分析验证p38MAPK信号通路抑制剂和葡萄糖氨对TNF-α诱导的IL-6、IL-8表达的影响。结果成功建立了TNF-α诱导的高表达IL-6、IL-8的血管内皮细胞损伤模型。壳寡糖可从mRNA转录和蛋白翻译水平抑制IL-6、IL-8的表达。初步揭示壳寡糖对TNF-α诱导血管内皮细胞表达IL-6、IL-8的抑制作用可能是通过p38的磷酸化和O-连接糖基化相互作用来完成的。结论壳寡糖可通过OGT调控TNF-α诱导的EA.hy926细胞IL-6、IL-8的表达,保护血管内皮细胞,减少炎症损伤。
Aim To investigate the protective effect of chitosan oligosaccharides（COS）in human vascular endothelial cells（EA.hy926） damaged by TNF-α.Method The model of cell treated with TNF-α to express high levels of IL-6 and IL-8 was established.The effect of COS on cell expression of IL-6 and IL-8 induced by TNF-α was estimated not only at the RNA transcriptional level but also in protein production through RT-PCR and ELISA assay,respectively.The effect of COS on activation of p38MAPK signaling pathways and expression of OGT in the cell was detected by Western blot.The effect of p38MAPK inhibitor and glucosamine on the expression of IL-6 and IL-8 induced by TNF-α at mRNA level was verified by RT-PCR.Results Model of injury cell which expressed high levels of IL-6 and IL-8 induced by TNF-α was established successfully.COS inhibited the expression of IL-6 and IL-8 induced by TNF-α at mRNA transcription and protein translation levels in the cell.The study revealted that the inhibitory effect of COS on vascular endothelial cells expression of IL-6 and IL-8 induced by TNF-α may be mediated via decreasing phosphorylation of p38MAPK and increasing O-GlcNAc modification of protein.Conclusion COS displayed a significant anti-inflammation effect through inhibiting IL-6 and IL-8 expression induced by TNF-α in EA.hy926 cells,which suggested that COS might have promising therapeutic potential against vascular diseases.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2011年第2期252-257,共6页
Chinese Pharmacological Bulletin
基金
重庆市自然科学重点项目(NoCSTC
2009BA5083)
重庆医科大学重点基金项目(NoXBED200806)
