本文選題：生物節(jié)律 + 雷帕霉素��； 參考：《山東醫(yī)藥》2017年42期

【摘要】：目的觀察雷帕霉素對(duì)衰老細(xì)胞生物節(jié)律關(guān)鍵基因的調(diào)控作用。方法通過(guò)傳代誘導(dǎo)人皮膚成纖維細(xì)胞復(fù)制性衰老,獲得衰老的人皮膚成纖維細(xì)胞。加入50%的馬血清刺激誘導(dǎo)細(xì)胞同步化,將細(xì)胞分為觀察組和對(duì)照組,分別加入100 mmol/L的雷帕霉素和DMSO培養(yǎng)24 h。采用熒光定量PCR法檢測(cè)生物節(jié)律關(guān)鍵基因芳烴受體核轉(zhuǎn)位蛋白3(BMAL1)、節(jié)律周期蛋白2(PER2)、藍(lán)光受體蛋白1(CRY1)、孤兒核受體α(Rev-erbα)、孤核受體α(Rorα)mRNA表達(dá),利用JTK-CYCLE軟件分析各基因的節(jié)律周期,Western blotting法檢測(cè)BMAL1、PER2、CRY1蛋白表達(dá)。結(jié)果觀察組PER2、CRY1、Rev-erbα、RorαmRNA表達(dá)高于對(duì)照組(P均0.05)。對(duì)照組BMAL1、CRY1、Rev-erbα、Rorα基因的節(jié)律周期為24 h,PER2基因的節(jié)律周期為28 h。觀察組BMAL1、CRY1、Rorα基因的節(jié)律周期為24 h,Rev-erbα基因的節(jié)律周期為27.5 h,PER2的節(jié)律周期為24 h。觀察組CRY1蛋白表達(dá)高于對(duì)照組(P0.05),PER2蛋白表達(dá)低于對(duì)照組(P0.05),兩組BMAL1 mRNA及蛋白表達(dá)比較無(wú)統(tǒng)計(jì)學(xué)差異。結(jié)論雷帕霉素可以提高衰老的人皮膚成纖維細(xì)胞生物節(jié)律關(guān)鍵基因BMAL1、PER2、CRY1、Rev-erbα、RorαmRNA的表達(dá),縮短PER2的節(jié)律周期,對(duì)紊亂的節(jié)律鐘功能具有調(diào)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 偽 (Rev-erb 偽) and solitary nucleus receptor 偽 (Ror 偽) 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 偽 and Ror 偽 in the observation group was higher than that in the control group (all P 0.05). In the control group, the Ror 偽 cycle of the CRY1A Rev-erb 偽 gene was 24 h and that of the PER2 gene was 28 h. In the observation group, the Ror 偽 gene cycle of CRY1or 偽 of BMAL1 was 24 h and that of Rev-erb 偽 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 偽 mRNA, shorten the rhythm cycle of PER2, and regulate the function of circadian clock in aged human skin fibroblasts.
【作者單位】： 深圳大學(xué);
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(81601215)
【分類號(hào)】：R33

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本文編號(hào)：2113319