摘要目的 探讨热处理、中波紫外线(UVB)辐射及两者联合作用下,体外培养的人表皮黑素细胞热休克蛋白72(heat shock protein 72,HSP72)的变化.方法 热处理(42℃,1 h/d,连续3d)、UVB辐射(50 mJ/cm2,连续3d)及两者联合(首先42℃,加热1h,然后50 m J/cm2 UVB照射,连续3d)处理正常人表皮(包皮)黑素细胞2h、6h后收集细胞,用实时荧光定量PCR技术和Western印迹技术分别检测不同组别间HSP72 mRNA及蛋白表达水平的差异.结果 热处理组(6.584±0.871)及联合作用组(7.269±0.454)与对照组(0.975±0.089)相比,mRNA表达水平明显增加(P＜ 0.001);UVB辐射组(0.832±0.084)与对照组相比,mRNA表达水平差异无统计学意义(P＞0.05).Western印迹结果显示,热处理组(2.022±0.058)及联合作用组(2.080±0.045 )HSP72蛋白表达水平明显高于对照组(0.532±0.033,P＜ 0.001 );UVB组(0.546±0.021)与对照组相比,表达水平差异无统计学意义(P＞0.05).析因设计方差分析结果显示,热处理与UVB辐射对HSP72 mRNA和蛋白表达均无交互作用(F=2.106,1.399,P＜ 0.05).结论 热处理引起HSP72表达显著增多,可能与热处理引起黑素细胞功能增强及UVB辐射后的损伤保护作用有关.

abstractsObjective To explore the effects of heat treatment and ultraviolet B (UVB) radiation alone or in combination on the expression of heat shock protein (HSP) 72 in human epidermal melanocytes.Methods Melanocytes were obtained from human foreskin,and subjected to primary culture.After 3 to 5 passages,the melanocytes were classified into 4 groups:control group (receiving no treatment),heat treatment group (treated with heat at 42 ℃ for 1 hour every day for 3 days),UVB group (irradiated with UVB at 50 mJ/cm2 daily for 3days),combination group (treated with heat at 42 ℃ for 1 hour followed by irradiation with UVB at 50 mJ/cm2daily for 3 days).After another 2- to 6-hour culture following the last treatment,melanocytes were collected and subjected to real time PCR and Western blot for the detection of HSP72 mRNA and protein expression,respectively.Results The mRNA and protein expressions of HSP72 were significantly higher in the heat treatment group and combination group than in the control group (mRNA:6.584 ± 0.871 and 7.269 ± 0.454 vs.0.975 ± 0.089,both P ＜ 0.001; protein:2.022 ± 0.058 and 2.080 ± 0.045 vs.0.532 ± 0.033,both P ＜ 0.001 ),but was similar between the UVB group and control group (mRNA:0.832 ± 0.084 vs.0.975 ± 0.089,P ＞ 0.05;protein:0.546±0.021 vs.0.532 ± 0.033,P ＞ 0.05).The ANOVA of factorial design showed that neither heat treatment nor UVB irradiation had interaction effect on the mRNA or protein expression of HSP72 (F =2.106,1.399 respectively,both P ＜ 0.05).Conclusions Heat treatment can cause an increase in the expression of HSP72,which may enhance the function of melanocytes and protect melanocytes from UVB induced damage.