目的:研究皮肤朗格汉斯(Langerhans)细胞(LC)在不同剂量中波紫外线(UVB)照射前、后p53蛋白磷酸化水平的差异。方法:联合采用密度梯度离心和免疫磁珠的方法分离纯化LC,将LC分为实验组和对照组,实验组分别接受30、60和90mJ/cm^2 UVB辐射,辐射后4h提取总蛋白,采用Western—blotting法检测各组细胞中p53蛋白及磷酸化p53蛋白的表达量,并比较两者之间的差异。结果:接受UVB辐射后,皮肤LC中p53蛋白表达基本无变化,而p53蛋白磷酸化水平显著升高,差异具有统计学意义,并呈剂量依赖性。结论:UVB辐射可刺激p53蛋白磷酸化,并呈剂量依赖性,进而诱导p53蛋白活化,发挥其下游功能。
Objective: To study the phosphorylated p53 protein expressions in the epidermal Langerhans cell (LC) with or without UVB irradiation. Methods: Density gradient centrifugation and magnetic cell sorting were used to separate and purify the LCs from human epidermis. LCs were divided into experimental group and control group. Different dosage UVB was irradiated to the experimental groups. The total proteins were harvested 4 hours after the application. Western-blotting was conducted to detect phosphorylated p53 protein expressions of each group. Results: After the UVB irradiation in different dosage, the phosphorylated p53 protein expressions in LCs were statistically higher than those in the control group (P〈 0.05), and were dosage dependent. Condusion: UVB irradiation increased the phosphorylated p53 protein expressions of epidermal LC and its pattern is dosage-dependent. So it may induce the activation of p53 to perform its functions consequently.