【摘要】 目的研究UVA对HaCaT细胞肿瘤坏死因子相关凋亡诱导配体(tumour necrosis factor related apoptosis inducing ligand,TRAIL)表达的影响;复制UVA诱导的HaCaT细胞凋亡模型,探究TRAIL在UVA诱导的HaCaT细胞凋亡中的作用及扇贝多肽(Polypeptide from Chlamys farreri,PCF)对UVA诱导的TRAIL凋亡通路的影响。方法实验设计分为5组:对照组、UVA模型组、UVA+5.69mmol·L-1PCF组、UVA+2.84mmol·L-1PCF组、UVA+1.42mmol·L-1PCF组。Real-TimePCR检测TRAIL mRNA表达;蛋白质印迹法检测TRAIL蛋白表达及caspase-8活性;琼脂糖凝胶电泳分析TRAIL中和性抗体对UVA诱导的HaCaT细胞凋亡的影响。结果8J·cm-2UVA照射HaCaT细胞后TRAIL mR-NA及蛋白表达增加,与对照组相比差异有显著性(P<0.01);TRAIL中和性抗体对UVA诱导的HaCaT细胞凋亡有抑制作用;1.42~5.69mmol·L-1剂量范围内的PCF可剂量依赖性抑制UVA引起的HaCaT细胞TRAIL mRNA及蛋白表达(P<0.05,P<0.01);PCF对UVA引起的HaCaT细胞caspase-8的活化有抑制作用,且呈量效关系。结论UVA可增强HaCaT细胞TRAIL表达;TRAIL参与了UVA诱导的HaCaT细胞凋亡;PCF对UVA诱导的HaCaT细胞TRAIL表达有抑制作用,也可减弱UVA诱导的caspase-8活化,以其抗氧化活性抑制TRAIL凋亡通路而发挥抗凋亡作用。更多还原

【Abstract】 Aim To investigate the impact of UVA on expression of tumour necrosis factor related apoptosis inducing ligand (TRAIL) and study the role of TRAIL in UVA-induced apoptosis of HaCaT cells as well as the influence of Polypeptide from Chlamys farreri(PCF)on TRAIL apoptotic pathway induced by UVA.Methods Cells were divided into five groups: control group, UVA model group,UVA+5.69 mmol·L-1 PCF group, UVA+2.84 mmol·L-1 PCF group, UVA+1.42 mmol·L-1 PCF group.Expression level of TRAIL mRNA was assayed by Real-Time PCR.Western blot analysis was used to determine the protein level of TRAIL and caspase-8 activation. The effect of TRAIL neutralization antibody on UVA-induced apoptosis was also investigated.Results TRAIL mRNA and protein levels increased after 8 J·cm-2 UVA radiation and the discrepancy was significant compared with control group(P<0.01).TRAIL neutralization antibody had inhibitory effect on UVA-induced apoptosis of HaCaT cells.1.42~5.69 mmol·L-1 PCF inhibited UVA-induced expression of TRAIL in HaCaT cells dose-dependently(P<0.05,P<0.01).PCF attenuated UVA-induced caspase-8 activation in a dose-dependent manner.Conclusions The enhancement of TRAIL expression induced by UVA in HaCaT cells indicated the involvement of TRAIL in UVA-induced apoptosis.PCF could attenuate UVA-induced TRAIL expression and block TRAIL apoptotic pathway, which may be attributed to its anti-apoptotic effect.更多还原