西罗莫司诱导自噬对UVB辐照下HaCaT细胞增殖和凋亡的初步影响  

Effects of sirolimus on the proliferation and apoptosis in HaCaT cells irradiated by UVB

作　　者：任发亮[1] 黄丹[1] 陈旭[1] 郑云鹏[1] 陈崑[1] 鞠梅[1] 常宝珠[1] 李新宇[1] 顾恒[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所,江苏南京210042

出　　处：《临床皮肤科杂志》2013年第6期335-338,共4页Journal of Clinical Dermatology

基　　金：江苏省自然科学基金(BK2011129);国家自然科学基金面上项目(81171513);教育部博士学科点专项科研基金(2011110611004)资助项目

摘　　要：目的:研究西罗莫司诱导自噬对UVB致HaCaT细胞光损伤的干预作用。方法:设置6组,分别为空白组、二甲基亚砜(DMSO)组、25mJ/cm^2 UVB组、25 mJ/cm^2 UVB+西罗莫司组、50mJ/cm^2 UVB组、50mJ/cm^2 UVB+西罗莫司组。单丹(磺)酰戊二胺(MDC)法检测不同处理后HaCaT细胞的自噬并比较各组自噬水平。噻唑蓝(MTT)法检测不同处理后HaCaT细胞的增殖活性。Hochest、碘化丙啶(PI)染色观察HaCaT细胞的凋亡和坏死。结果:MDC染色结果可见西罗莫司处理组的自噬体阳性细胞百分率分别高于相应UVB照射组。MTT法检测到25 mJ/cm^2 UVB+西罗莫司组的细胞增殖抑制率低于25 mJ/cm^2 UVB组,而50mJ/cm^2 UVB+西罗莫司组高于50 mJ/cm^2 UVB组。Hochest染色观察到西罗莫司处理组的凋亡百分率分别低于相应UVB组。PI染色可见UVB照射组出现坏死细胞,而西罗莫司干预组未见坏死细胞。结论:西罗莫司诱导自噬可减轻25 mJ/cm^2 UVB辐照对HaCaT细胞增殖能力的抑制,而在50mJ/cm^2 UVB辐照下加重对细胞的抑制。西罗莫司诱导自噬可明显减轻25、50mJ/cm^2 UVB辐照下HaCaT细胞的凋亡和坏死,对UVB损伤有保护作用。Objective： To study the effects of sirolimus-induced autophagy on UVB-irradiation induced HaCaT cell damaged. Methods： HaCaT cells were divided into 6 groups, each of which was treated with vehicle, sirolimus, UVB or combination of sirolimus and UVB. MDC and MTT assays were used to measure cell autophagosome levels and proliferation respectively. Apoptosis and necrosis in HaCaT cells were assessed by Hoechest and PI staining respectively. Results： The percentage of the autopagosome-positive cells in sirolumus treated group was higher than that in UVB irradiated cells. In comparison with U- VB irradiation alone, combination of sirolimus and 25 mJ/cm2 UVB irradiation induced less inhibition of proliferation. In corn parison with 50 mJ/cm2 UVB irradiation alone, combination of sirolimus and 50 mJ/cm2 UVB irradiation induced a higher growth inhibition rate. Hoechest staining showed that apoptosis rate in sirolimus-treated cells was lower than that in UVB irradiated cells. Moreover, no necrotic cells were seen in sirolimus-treated groups while some necrotic ceils were observed following UVB irradiation. Conclusions： Sirolimus induced autophagy can attenuate HaCaT cell growth inhibition caused by 25 mJ/cm2 UVB irradiation. In contrast, sirolimus enhances growth inhibition induced by 50 mJ/cm2 UVB irradiation. Sirolimus induced au- tophagy can decrease HaCaT cell apoptosis and necrosis induced by UVB irradiation at the doses of 25 or 50 mJ/cm2, and protect HaCaT ceils against UVB irradiation.

关 键 词：角质形成细胞 自噬 西罗莫司 紫外线 中波 

分 类 号：R339.57[医药卫生—人体生理学]