雷帕霉素抑制光老化成纤维细胞周期阻滞的研究

Inhibiting Effect of Rapamycin on the Cell Cycle Arrest of Photoaged Fibroblasts

目的探讨雷帕霉素(rapamycin,RAPA)对光老化成纤维细胞(fibroblasts,FBs)周期及相关蛋白的影响。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UVB体外反复照射构建FBs的老化模型,流式细胞技术检测各组周期的分布变化,Western Blot技术检测细胞周期相关蛋白p53和p21的表达。结果低浓度RAPA对细胞活性并无影响。与UVB照射组比较,RAPA+UVB组的S期细胞比例下降了6.5%,细胞周期相关蛋白p53和p21的表达分别下降了26.3%和27.8%。结论 RAPA可以缓解光老化FBs的S期阻滞,降低周期蛋白p53和p21的产生,一定程度上改善UVB照射导致的细胞衰老。

Objective To explore the effect of rapamycin （RAPA） on the cell cycle of photoaged fibroblasts （FBs）and on the expressions of associated proteins. Methods Cell counting kit-8 （CCK-8） assay was applied in detecting the effectof RAPA of different concentration on the activity of FBs so as to find out the most suitable concentration; the aged model ofFBs was built up by repeated UVB radiation; flow cytometry was applied in detecting the changes of cell cycle distributionwhile Western Blot Analysis in detecting the expressions of proteins related to cell cycle： protein p53 and protein p21. ResultsRAPA of low concentration was of no effect on cell activity; compared with FBs radiated by UVB, the cell ratio in S phaseof FBs pretreated by RAPA and UVB decreased 6.5%, the expressions of protein p53 and protein p21 decreased 26.3% and27.8% respectively. Conclusions RAPA can relieve the S-phase arrest of photoaged FBs, decrease the expression of proteinp53 and protein p21, and thus improve the cell aging induced by UVB radiation to some extent.