肿瘤坏死因子相关凋亡诱导配体及扇贝多肽在UVA诱导HaCaT细胞凋亡中的作用

The role of tumor necrosis factor related apoptosis inducing ligand and Polypeptide from Chlamys farreri in UVA-induced apoptosis in HaCaT cells

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-Time PCR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 mRNA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。

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 with control group （ P 〈 0.01 ）. antibody had inhibitory effect on sis of HaCaT cells. 1.42 - 5.69 significant compared TRAIL neutralization UVA-induced apoptommol · 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 dosedependent 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.