摘要
目的探讨雷帕霉素(RAPA)对紫外线照射导致的成纤维细胞(FBs)衰老和抗氧化应激的作用特点。方法采用CCK-8法检测不同浓度的RAPA对FBs活性的影响,甄选最佳应用浓度。利用紫外线体外反复照射构建成纤维细胞的老化模型。RAPA作用后,β-半乳糖苷酶染色法检测衰老细胞,利用DCFH-DA为荧光探针,用流式细胞仪检测细胞内活性氧簇(ROS)的水平。结果低浓度的RAPA对细胞的活性并无影响;与紫外线照射组组比较,RAPA+紫外线组的β-半乳糖苷酶染色阳性率下降了约40%,差异具有统计学意义(P<0.05)。RAPA预处理可以明显降低紫外线照射细胞组ROS的产生。结论 RAPA可以降低光老化成纤维细胞ROS生成量,一定程度上缓解了紫外线照射导致的细胞光老化进程。
Objective To explore the influence of RAPA on photo-aging and ant-oxidative stress of fibroblasts（FBs） induced by repeated exposure to ultraviolet B（UVB）. Methods The effect of different concentrations of RAPA on FBs proliferation was measured by cell counting kit-8（CCK-8） assay, and the optimum concentration was selected. The photo-aging model of FBs was induced by repeated UVB irradiation. After incubation with RAPA, the senescent cells were detected with β-galactosidase staining. The intracellar level of ROS was measured by flow cytometry（FCM） with DCFH-DA as fluorescent probe. Results The low concentration of RAPA had no effect on fibroblast cell viability or proliferation. Compared with the UVB irradiation group, the positive rate of β-galactosidase staining decreased about 40% in the UVB ＋ RAPA group. The difference was statistically significant（P〈0.05）. The production of ROS in the UVB irradiated cells reduced significantly after RAPA pretreatment. Conclusion Pretreatment with RAPA can decrease the expression of ROS and inhibit UVB-induced photo-aging process to a certain extent.
出处
《中国美容整形外科杂志》
CAS
2017年第8期499-501,507,共4页
Chinese Journal of Aesthetic and Plastic Surgery
