Tempol对紫外线照射所致HaCaT细胞损伤保护作用  

PROTECTION OF TEMPOL AGAINST UVB IRRADIATION-INDUCED DAMAGE ON HaCaT CELLS

作　　者：于洋[1] 王国英[2] 

机构地区：[1]青岛大学医学院附属医院皮肤科,山东青岛266021 [2]青岛大学医学院免疫学教研室,山东青岛266021

出　　处：《齐鲁医学杂志》2009年第4期327-329,332,共4页Medical Journal of Qilu

摘　　要：目的观察氮氧化物4-羟基-2,2,6,6-四甲基哌啶(Tempol)对中波紫外线(UVB)照射所致人角质形成细胞(HaCaT细胞)损伤的保护作用,并探讨其发生机制。方法在HaCaT细胞培养系统中加入不同浓度的Tempol进行培养,12h后洗掉培养液中的Tempol,用30mJ/cm2UVB照射细胞后,重新加入培养液继续培养24h。用MTT法检测细胞增殖活性,流式细胞术测定细胞凋亡率。RT-PCR方法测定FoxO3a和BubR1基因的表达。结果UVB照射组与正常对照组相比,细胞增殖活性明显降低(F=27.71,q=13.57,P<0.05),FoxO3amRNA的表达明显增强(F=13.93,q=9.79,P<0.05),BubR1mRNA的表达明显减弱(F=29.69,q=14.54,P<0.05),凋亡率明显增高(χ2=93.69,P<0.05);与UVB照射组相比,不同浓度的Tempol可以恢复UVB照射细胞的增殖能力(q=3.24～13.60,P<0.05),降低照射细胞的凋亡率(χ2=12.02～101.02,P<0.05),下调FoxO3amRNA的表达(q=2.92～9.95,P<0.05),上调BubR1mRNA的表达(q=2.97～14.61,P<0.05)。结论Tem-pol可以保护HaCaT细胞免受UVB照射造成的损伤。Objective To study the protection of nitroxide 4-hydroxy-2,2,6,6,-tetramethyl piperidin （Tempol） against UVB irradiation-induced damage on cultured HaCaT cells and its mechanism. Methods HaCaT cells were cultured with different concentrations of Tempol. After 12 hours of incubation, Tempol was washed away, and the HaCaT cells were irradiated with UVB for four seconds followed by 24 hours of incubation with fresh medium. Cell proliferation ability was studied with MTT assay and cell apoptosis analyzed by flow cytometry. Gene expressions of FoxO3a and BubR1 were detected by RT-PCR. Results Com pared with the control, in UVB group, HaCaT cell proliferation activity was suppressed （F= 27. 71 ,q= 13.57,P〈0. 05）, the expression of FoxO3a mRNA was significantly increased （F= 13.93,q=9.79 ,P〈0.05） ; the apoptoic rate was significantly enhanced （x^2 =93.69,P〈0. 05） and BubR1 mRNA was significantly decreased （F=29.69,q=14.54,P〈0.05）. Compared with the UVB group, different concentrations of Tempol could significantly increase the proliferation ability （q=3. 24-13.60,P〈0. 05） and decrease the apoptotic rate of UVB-irradiated HaCaT cells （x^2 = 12.02- 101.02, P〈0.05）. Furthermore, Tempol could significantly down-regulate the mRNA expression of FoxO3a （q=2. 92-9. 95 ,P〈0.05） and significantly up-regulate the expression of BubR1 mRNA （q=2.97-14. 61,P〈0.05） of UVB-irradiated HaCaT cells. Conclusion Tempol could protect HaeaT cells against UVB irradiation-induced cell damage.

关 键 词：紫外线 辐射损伤 实验性 环氮氧化物 HACAT细胞 细胞凋亡 FoxO3a基因 BubR1基因 

分 类 号：R122.25[医药卫生—环境卫生学]