中长波紫外线辐射诱导皮肤成纤维细胞上清液中微囊泡功能的检测  

Function of UVA/UVB-induced Microvesicles in Human Skin Fibroblasts

作　　者：刘娟[1] 周炳荣[1] 王申[1] 张家安[1] 张丽超[1] 吴红巾 易飞[1] 谢淑芬[1] 范曾[1] 骆丹[1] 

机构地区：[1]南京医科大学第一附属医院皮肤性病科,江苏南京210029

出　　处：《中国皮肤性病学杂志》2016年第1期1-5,共5页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金(81371757;81573072)

摘　　要：目的观察中长波紫外线辐射诱导皮肤成纤维细胞微囊泡的生成及其对人皮肤成纤维细胞形态及增殖活性的影响。方法使用不同剂量中长波紫外线辐射人皮肤成纤维细胞，提取细胞上清液中的微囊泡，利用透射电镜及Western免疫印迹法鉴定所获得的微囊泡，BCA法测定微囊泡蛋白的含量；将紫外线辐射后生成的微囊泡与正常成纤维细胞共孵育，使用倒置显微镜观察细胞形态，并使用细胞计数CCK-8法检测细胞增殖率。结果中长波紫外线辐射皮肤成纤维细胞促进微囊泡的生成和释放。紫外线辐射后的微囊泡直径多数在20～200nm之间，表面蛋白Alix，tsg101及CD9表达阳性。紫外线辐射后生成的微囊泡可导致人皮肤成纤维细胞形态发生改变，倒置显微镜下出现较多的细胞碎片，细胞体积变大，过度伸展，并使细胞增殖率降低。结论中长波紫外线辐射可诱导皮肤成纤维细胞释放微囊泡，后者是介导紫外线辐射旁效应的主要介质之一。Objective To observe the production of microvesicles, and the effect of cell morphology and proliferation activity of human skin fibroblasts with acute UVA/UVB radiation. Methods Apply different doses of UVA/ UVB irradiation on human skin fibroblasts, and extract microvesicles in the supernatant fluid. Use transmission electron microscopy and Western blot to identify the microvesicles, and BCA method to determine the protein of microvesicles, ncubate the nomal fibroblasts with microvesicles induced by the ultraviolet radiation. Then the cell morphology was recorded using an inverted microscope and cell proliferation rate was de- tected by cell counting CCK-8 method. Results Acute UV radiation can promote the production and release of microvesicles in skin fibroblasts. After UV radiation, the majority diameter of microvesicles were between 20 -200hm,The surface proteins Alix, TSG101 and CD9 were positive. After UV radiation, the production of microvesicles can lead to morphology change of human skin fibroblast. Under the microscope, the cell debris increased, the cell volume were larger and the cells over extended. The cell proliferation rate were reduced as well. Conclusion Acute UVA radiation can induce skin fibroblasts to release microvesicles, and the latter is one of the main medium mediating the bystander effect of ultraviolet radiation.

关 键 词：中长波紫外线 成纤维细胞 微囊泡 细胞老化 

分 类 号：Q813.1[生物学—生物工程] R329.2[医药卫生—人体解剖和组织胚胎学]