【摘要】：目的研究PI3K-Akt-mTOR信號通路對Foxp3基因表達(dá)的影響。方法將SPF級昆明系小鼠60只隨機(jī)分為對照組(A)和實(shí)驗(yàn)組(B、C、D),B、C、D三組分別灌胃雷帕霉素0.2、0.4、0.6 mg.d-1,A組每天予以無菌水灌胃,共3周。3周后,無菌條件下心臟采血,EDTA抗凝,分離脾臟,制備單細(xì)胞懸液,采用流式細(xì)胞儀檢測小鼠外周血和脾臟中CD4+CD25+調(diào)節(jié)性T細(xì)胞水平(CD4+CD25+Treg細(xì)胞占CD4+T細(xì)胞的百分比),RT-PCR檢測小鼠脾細(xì)胞Foxp3mRNA的表達(dá),免疫組織化學(xué)SP法染色觀察小鼠脾臟中p-mTOR蛋白表達(dá)的變化,并利用計(jì)算機(jī)圖像分析技術(shù)測量各實(shí)驗(yàn)組及對照組中p-mTOR蛋白表達(dá)的平均光密度。結(jié)果 Foxp3 mRNA在實(shí)驗(yàn)組(B、C、D)小鼠脾臟中表達(dá)的程度分別為(0.4853±0.0574、0.7886±0.1085、0.9639±0.2015),明顯高于對照組A(0.1345±0.0271)(P0.05);實(shí)驗(yàn)組(B、C、D)小鼠脾臟中p-mTOR蛋白表達(dá)的平均光密度分別為(0.2326±0.0431、0.1156±0.0223、0.0556±0.0041),明顯低于對照組A(0.4223±0.0534)(P0.05);兩指標(biāo)經(jīng)pearson相關(guān)分析,小鼠脾臟中Foxp3mRNA的表達(dá)與p-mTOR蛋白表達(dá)呈負(fù)相關(guān)。結(jié)論抑制PI3K-Akt-mTOR信號通路會促使Foxp3的表達(dá)增強(qiáng);PI3K-Akt-mTOR信號通路的激活程度與Foxp3的表達(dá)程度呈負(fù)相關(guān)。
[Abstract]:Objective to study the effect of PI3K-Akt-mTOR signaling pathway on Foxp3 gene expression. Methods 60 Kunming mice of SPF grade were randomly divided into two groups: control group (A) and experimental group (group D). After 3 weeks, blood was collected from the heart, EDTA was anticoagulant, spleen was isolated and single cell suspension was prepared under aseptic condition. The levels of CD4 CD25 regulatory T cells (the percentage of CD4 CD25 Treg cells in CD4 T cells) in peripheral blood and spleen of mice were detected by flow cytometry, and the expression of Foxp3mRNA in spleen cells of mice was detected by RT-PCR. Immunohistochemical SP staining was used to observe the expression of p-mTOR protein in the spleen of mice, and the average optical density of p-mTOR protein expression in each experimental group and control group was measured by computer image analysis. Results the expression of Foxp3 mRNA in the spleen of the experimental group was (0.4853 鹵0.0574) 0.7886 鹵0.1085 鹵0.9639 鹵0.2015, which was significantly higher than that of the control group A (0.1345 鹵0.0271) (P0.05). The average optical density of p-mTOR protein expression in the spleen of the experimental group was (0.2326 鹵0.0431) 0.1156 鹵0.022 3n 0.0556 鹵0.0041, which was significantly lower than that of the control group A (0.4223 鹵0.0534) (P0.05). By pearson correlation analysis, the expression of Foxp3mRNA in mouse spleen was negatively correlated with the expression of p-mTOR protein. Conclusion inhibition of PI3K-Akt-mTOR signaling pathway can increase the expression of Foxp3, and the activation of PI3K-Akt-mTOR signaling pathway is negatively correlated with the expression of Foxp3.
【作者單位】： 安徽醫(yī)科大學(xué)第一附屬醫(yī)院心臟血管外科;
【分類號】：R-332

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