miR-126基因敲减对小鼠胸腺淋巴细胞发育的影响

发布时间：2018-02-27 12:39

本文关键词： 微小RNA- 基因敲减胸腺 CD+T细胞　出处：《基础医学与临床》2016年03期 　论文类型：期刊论文

【摘要】：目的研究miR-126基因敲减对小鼠胸腺细胞发育的影响,并初步探讨其意义。方法观察miR-126基因敲减(KD)小鼠胸腺的体积、重量和细胞总数;Real-time PCR检测小鼠胸腺miR-126表达水平;HE染色观察小鼠胸腺形态学结构;FACS检测胸腺中淋巴细胞比例、核抗原Ki-67的表达以及细胞凋亡的变化;最后,Western blot检测胸腺中磷酸化AKT(p-AKT)和磷酸化ERK1/2(p-ERK)的表达。结果 WT和miR-126KD小鼠胸腺体积大小、重量以及细胞总数均无明显差异,但miR-126KD小鼠胸腺miRNA-126表达水平显著下调(P0.05)。miR-126KD小鼠胸腺组织结构出现形态学异常。与WT小鼠相比,miR-126KD小鼠胸腺CD4~+SP细胞比例及细胞绝对数明显增加(P0.05),而CD4~+CD8~+(DP)细胞比例及绝对数则显著减少(P0.05),miR-126KD小鼠胸腺CD4~+SP细胞核抗原Ki-67表达显著增加(P0.05),且细胞凋亡明显减少(P0.05),而DP细胞Ki-67表达则明显减少(P0.05)。最后,miR-126KD小鼠胸腺淋巴细胞中磷酸化AKT和磷酸化ERK1/2的表达水平明显下调(P0.05)。结论 miR-126基因敲减可显著影响胸腺细胞特别是CD4~+SP细胞的发育,为后续进一步深入探讨miR-126对T淋巴细胞发育以及功能的影响提供重要的实验基础。
[Abstract]:Objective to study the effect of miR-126 knockout on the development of thymocytes in mice and its significance. Methods the volume of thymus in miR-126 knockout mice was observed. The expression level of miR-126 in thymus of mice was detected by real-time PCR. The morphological structure of thymus was observed by HE staining. The percentage of lymphocytes in thymus, the expression of nuclear antigen Ki-67 and the changes of apoptosis were observed. Finally, we detected the expression of phosphorylated AKTa-p-AKT and phosphorylated ERK1 / 2p-ERK in thymus by Western blot. The results showed that there was no significant difference in the volume, weight and total number of cells between WT and miR-126KD mice. However, the expression of miRNA-126 in thymus of miR-126KD mice was significantly down-regulated. The morphologic changes of thymus tissue were observed in P0.05, miR-126KD mice. Compared with WT mice, the percentage and absolute number of CD _ 4 ~ SP cells in thymus of miR-126KD mice were significantly increased, while the percentage of CD _ 4 ~ CD _ (8) ~ (DPN) cells and the absolute number of cells in thymus were significantly increased compared with WT mice. Logarithm significantly decreased the expression of CD _ 4 ~ SP nuclear antigen Ki-67 in thymus of P0.05 ~ 126KD mice, and significantly decreased the apoptosis of P0.05, while the expression of Ki-67 in DP cells decreased the expression of P0.05. Finally, phosphorylated AKT and phosphorylated AKT in thymocytes of miR-126KD mice were significantly reduced. Conclusion miR-126 knockout can significantly affect the development of thymocytes, especially CD4 ~ SP cells. It provides an important experimental basis for the further study of the effect of miR-126 on the development and function of T lymphocytes.
【作者单位】：遵义医学院免疫学教研室贵州省免疫学研究生教育创新基地;
【基金】：国家自然科学基金(31370918) 教育部新世纪优秀人才计划(NCET-12-0661)
【分类号】：R392

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