小鼠CD4+ T细胞活化后4种microRNAs的表达分析

目的: 研究小鼠脾脏CD4⁺ T淋巴细胞活化后微小RNA(microRNA，miRNA)的表达水平。方法: 采用CD3/CD28 免疫磁珠法刺激培养小鼠脾脏CD4⁺T细胞24 h，应用流式细胞术检测刺激前后CD4⁺T细胞的活化率，使用实时荧光定量PCR技术分别检测培养0 h，6 h，12 h以及24 h后的CD4⁺T细胞中miR-181d、miR-342-3p、miR-9、miR-122-3p的表达水平。结果： CD3/CD28 免疫磁珠法刺激培养24 h后CD4⁺ T细胞活化率由5.2%升高至60.9%，差异有统计学意义（t=22.01，P <0.01）。与培养0 h相比，miR-181d和miR-342-3p在经刺激培养6 h，12 h和24 h后的CD4⁺T细胞中的表达水平均明显降低（P <0.05），而miR-9和miR-122-3p的表达各组间的差异无统计学意义（P>0.05）。结论： 小鼠脾脏CD4+T细胞活化后，miR-181d和miR-342-3p的表达明显下调。

Analysis of four microRNAs expression in activated mouse CD4+T lymphocytes

Objective: To explore the expression level of four microRNAs(miRNA) in activated CD4⁺T lymphocytes from mouse spleen. Methods: CD4⁺T cells were isolated from mouse spleen and stimulated with anti-CD3/CD28 beads for 24 h. The activity index of CD4+T cells was determined by flow cytometry at 0 h and 24 h after stimulation, and the relative expression of miR-181d, miR-342-3p, miR-9 and miR-122-3p were quantified by real-time PCR at 0 h, 6 h, 12 h and 24 h after stimulation, respectively. Results: Anti-CD3/CD28 beads could activate CD4⁺T lymphocytes, and the activity index of CD4⁺T cells significantly increased from 5.2% to 60.9%(t=22.01, P<0.01). Expression levels of miR-181d and miR-342-3p in stimulation group at 6 h, 12 h, and 24 h decreased compared with pro-stimulation group(P<0.05). Difference of miR-9 and miR-122-3p expression between stimulation and pre stimulation group did not reach significance(P>0.05). Conclusion: Expressions of miR-181d and miR-342-3p were inhibited in activated CD4⁺T lymphocytes from mouse spleen.