雷帕霉素对衰老细胞生物节律关键基因的调控作用观察

发布时间：2018-07-10 11:55

本文选题：生物节律 + 雷帕霉素　；参考：《山东医药》2017年42期

【摘要】：目的观察雷帕霉素对衰老细胞生物节律关键基因的调控作用。方法通过传代诱导人皮肤成纤维细胞复制性衰老,获得衰老的人皮肤成纤维细胞。加入50%的马血清刺激诱导细胞同步化,将细胞分为观察组和对照组,分别加入100 mmol/L的雷帕霉素和DMSO培养24 h。采用荧光定量PCR法检测生物节律关键基因芳烃受体核转位蛋白3(BMAL1)、节律周期蛋白2(PER2)、蓝光受体蛋白1(CRY1)、孤儿核受体α(Rev-erbα)、孤核受体α(Rorα)mRNA表达,利用JTK-CYCLE软件分析各基因的节律周期,Western blotting法检测BMAL1、PER2、CRY1蛋白表达。结果观察组PER2、CRY1、Rev-erbα、RorαmRNA表达高于对照组(P均0.05)。对照组BMAL1、CRY1、Rev-erbα、Rorα基因的节律周期为24 h,PER2基因的节律周期为28 h。观察组BMAL1、CRY1、Rorα基因的节律周期为24 h,Rev-erbα基因的节律周期为27.5 h,PER2的节律周期为24 h。观察组CRY1蛋白表达高于对照组(P0.05),PER2蛋白表达低于对照组(P0.05),两组BMAL1 mRNA及蛋白表达比较无统计学差异。结论雷帕霉素可以提高衰老的人皮肤成纤维细胞生物节律关键基因BMAL1、PER2、CRY1、Rev-erbα、RorαmRNA的表达,缩短PER2的节律周期,对紊乱的节律钟功能具有调控作用。
[Abstract]:Objective to observe the regulation of rapamycin on key genes of biological rhythm in aging cells. Methods Human skin fibroblasts were induced to replicate senescence by subculture, and human skin fibroblasts were obtained. The cells were induced to synchronize by adding 50% equine serum. The cells were divided into observation group and control group. The cells were cultured with 100 mmol / L rapamycin and DMSO for 24 h, respectively. The mRNA expressions of aromatic receptor nuclear translocation protein 3 (BMAL1), rhythm cyclin 2 (PER2), blue light receptor protein 1 (CRY1), orphan nuclear receptor 伪 (Rev-erb 伪) and solitary nucleus receptor 伪 (Ror 伪) were detected by fluorescence quantitative PCR. JTK-CYCLE software was used to detect the expression of BMAL1 / PER2 / CRY1 protein by Western blotting. Results the mRNA expression of PER2, CRY1, Rev-erb 伪 and Ror 伪 in the observation group was higher than that in the control group (all P 0.05). In the control group, the Ror 伪 cycle of the CRY1A Rev-erb 伪 gene was 24 h and that of the PER2 gene was 28 h. In the observation group, the Ror 伪 gene cycle of CRY1or 伪 of BMAL1 was 24 h and that of Rev-erb 伪 was 27.5 h and that of PER2 was 24 h. The expression of CRY1 protein in the observation group was higher than that in the control group (P0.05) and the expression of PER2 protein was lower than that in the control group (P0.05). There was no significant difference in the expression of BMAL1 mRNA and protein between the two groups. Conclusion Rapamycin can increase the expression of Ror 伪 mRNA, shorten the rhythm cycle of PER2, and regulate the function of circadian clock in aged human skin fibroblasts.
【作者单位】：深圳大学;
【基金】：国家自然科学基金资助项目(81601215)
【分类号】：R33

【相似文献】