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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。
Objective To investigate the protective mechanism of chitosan oligosaccharide on vascular endothelial cell injury. Methods EA.hy926 vascular endothelial cells were used as experimental subjects. Endothelial cell injury models were established by TNF-α stimulation to express IL-6 and IL-8. The effects of chitooligosaccharides on the expression of IL-6 and IL-8 were detected by RT-PCR and ELISA from mRNA and protein levels respectively. Western blot was used to detect the effect of chitooligosaccharides on p38MAPK signal pathway and glycosyltransferase (OGT) expression. The effects of p38 MAPK signaling pathway inhibitor and glucosamine on TNF-α-induced IL-6 and IL-8 expression were analyzed. Results The model of vascular endothelial cell injury induced by TNF-α with high expression of IL-6 and IL-8 was successfully established. Chitooligosaccharides can inhibit the expression of IL-6 and IL-8 from mRNA transcription and protein translation level. The preliminary study revealed that the inhibitory effect of chito-oligosaccharides on TNF-α-induced expression of IL-6 and IL-8 in vascular endothelial cells may be through the phosphorylation of p38 and O-linked glycosylation interactions. Conclusion Chitosan oligosaccharide can regulate the expression of IL-6 and IL-8 in EA.hy926 cells induced by TNF-α by OGT and protect the vascular endothelial cells and reduce the inflammatory injury.