苦荞蛋白源肽AFYRW对脂多糖诱导的人脐静脉内皮细胞损伤的影响

Ameliorative Effect of Peptide AFYRW Derived from Tartary Buckwheat Protein on Lipopolysaccharide-Induced Injury in Human Umbilical Vein Endothelial Cells

目的:探讨苦荞蛋白源肽AFYRW在脂多糖(lipopolysaccharide,LPS)诱导的血管内皮细胞损伤中的作用。方法:采用LPS诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)建立血管内皮细胞炎症损伤模型;采用CCK8试剂盒检测AFYRW对细胞增殖活力的影响;Western blot检测血管细胞黏附分子-1(vascular cell adhension molecule-1,VCAM-1)、细胞内黏附分子-1(intercellular adhension molecule-1,ICAM-1)、白细胞介素(interleukin,IL)-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、内皮型一氧化氮合酶(endothelial nitric oxide synthase,e NOS)、诱导型一氧化氮合酶(inducible nitricoxide synthase,iNOS)水平及核因子-κB(nuclear factor kappa-B,NF-κB)p65的磷酸化水平;酶法检测一氧化氮(nitric oxide,NO)的含量变化;单核细胞黏附实验检测HUVECs的黏附性。结果:AFYRW明显提高了细胞的生存活力,降低VCAM-1、ICAM-1、IL-6、TNF-α、iNOS蛋白的相对表达量,减少了NO含量和单核细胞对内皮细胞的黏附,抑制NF-κB的活性。结论:AFYRW对LPS诱导的HUVECs炎症具有改善作用,其机制可能与抑制NF-κB的活性有关。

Objective:To investigate the ameliorative effect of peptide AFYRW derived from tartary buckwheat protein against LPS-induced vascular endothelial injury.Methods:Lipopolysaccharide(LPS)was used to induce vascular endothelial injury in human umbilical vein endothelial cells(HUVECs).The cell counting kit-8(CCK8)assay was used to detect the effect of AFYRW on cell proliferation.Western blot was used to evaluate the protein expression of vascular cell adhesion molecule-1(VCAM-1),intracellular adhesion molecule-1(ICAM-1),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),endothelial nitric oxide synthase(eNOS),inducible nitric oxide synthase(iNOS),and phosphorylation of nuclear factor-κB(NF-κB)p65.An enzymatic assay was used to detect the changes in the content of nitric oxide(NO).Monocyte adhesion assay was used to detect the adhesiveness of HUVECs.Results:Peptide AFYRW significantly increased the survival activity of cells,decreased the relative protein expression of VCAM-1,ICAM-1,IL-6,TNF-α,and iNOS,reduced the content of NO,inhibited LPS-induced adhesion of human monocyte THP-1 cells to HUVECs and suppressed the activity of NF-κB.Conclusion:Peptide AFYRW can ameliorate LPS-induced inflammation in HUVECs,which may be related to the inhibition of NF-κB activity.