| miR-125a-5p对实验性自身免疫性重症肌无力小鼠胸腺Treg细胞的影响 |
| 投稿时间：2025-02-27 修订日期：2025-04-06 点此下载全文 |
| 引用本文：谭舒婷,黎入荧,陈柳伶,刘竞丽,李劲频.miR-125a-5p对实验性自身免疫性重症肌无力小鼠胸腺Treg细胞的影响[J].医学研究杂志,2025,54(8):34-41 |
| DOI：
10.11969/j.issn.1673-548X.2025.08.007 |
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| 基金项目:国家自然科学基金资助项目(81960241) |
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| 中文摘要:目的 探讨miR-125a-5p对实验性自身免疫性重症肌无力(experimental autoimmune myasthenia gravis,EAMG)小鼠胸腺调节性T细胞(regulatory T cell,Treg)的影响。方法 使用鼠源性乙酰胆碱受体α亚基97~116多肽免疫诱导建立EAMG模型,随机分为对照组和EAMG组,对小鼠进行Lennon评分和低频重复电刺激实验评估模型。为进一步探讨miR-125a-5p对Treg细胞的影响,分别给予EAMG小鼠尾静脉注射阴性对照腺病毒和miR-125a-5p抑制腺病毒,实验分组为阴性对照组和miR-125a-5p抑制组。采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)法检测miR-125a-5p和叉状头转录因子P3(forkhead box P3,Foxp3)mRNA表达水平,Western blot法检测Foxp3蛋白表达水平,流式细胞术检测Treg细胞数量百分比,酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法检测血清白细胞介素-10(interleukin 10,IL-10)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)表达水平。结果 成功构建EAMG小鼠模型,与对照组比较,EAMG组Lennon评分升高,5Hz低频重复电刺激阳性,miR-125a-5p表达升高,Foxp3mRNA和蛋白表达均下降,Treg细胞数量百分比减少,IL-10、TGF-β1表达降低；抑制EAMG小鼠miR-125a-5p表达后,与阴性对照组比较,miR-125a-5p抑制组miR-125a-5p表达降低,Lennon评分降低,Foxp3mRNA和蛋白表达均升高,Treg细胞数量百分比增加,IL-10、TGF-β1表达升高,差异均有统计学意义(P<0.05)。结论 miR-125a-5p在EAMG中表达上调,抑制miR-125a-5p表达能够上调Foxp3的表达,促进Treg细胞数量增加和升高IL-10、TGF-β1水平,减轻EAMG小鼠肌无力症状,表明miR-125a-5p可能通过影响Foxp3导致Treg细胞数量减少和功能异常参与重症肌无力疾病的发生。 |
| 中文关键词:重症肌无力 miR-125a-5p 调节性T细胞 |
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| Effect of miR-125a-5p on Thymic Treg Cells in Mice with Experimental Autoimmune Myasthenia Gravis. |
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| Abstract:Objective To explore the effect of miR-125a-5p on thymic regulatory T cell (Treg) in mice with experimental autoimmune myasthenia gravis (EAMG). Methods The EAMG model was established by immunoinduction with murine-derived acetylcholine receptor α subunit 97-116 peptide and randomly divided into control group and EAMG group, and the mice were subjected to Lennon scoring and low-frequency repetitive electrical stimulation experiments to evaluate the model. To further explore the effect of miR-125a-5p on Treg cells, EAMG mice were injected with negative control adenovirus and miR-125a-5p-inhibited adenovirus by tail vein injection respectively, and the experimental groups were the negative control group and miR-125a-5p inhibiting group. The mRNA expression levels of miR-125a-5p and forkhead box protein P3 (Foxp3) were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), while the protein expression level of Foxp3 was detected by Western blot. The percentage of the number of Treg cells was detected by flow cytometry, and serum concentrations of interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) were assessed using enzyme-linked immunosorbent assay (ELISA). Results The EAMG mouse model was successfully established. Compared to the control group, the EAMG group exhibited a significantly higher Lennon score, positive responses to low-frequency repetitive electrical stimulation at 5Hz, and an upregulated expression of miR-125a-5p, downregulated of Foxp3mRNA and protein, reduced percentage number of Treg cells, and decreased expression of IL-10 and TGF-β1. Upon inhibition of miR-125a-5p expression in EAMG mice, compared with the negative control group, the expression of miR-125a-5p in the miR-125a-5p inhibiting group was decreased, and the Lennon score was decreased, Foxp3mRNA and protein expression were increased, with a significant increase in the percentage of Treg cells and enhanced expression of IL-10 and TGF-β1, and the differences were all statistically significance (P<0.05). Conclusion The expression of miR-125a-5p was upregulated in EAMG. Inhibition of miR-125a-5p expression led to upregulation of Foxp3, an increase in the number of Treg cells, elevated levels of IL-10 and TGF-β1, and attenuation of myasthenia gravis symptoms in EAMG mice. These results suggest that miR-125a-5p may be involved in developing myasthenia gravis disease by affecting Foxp3, leading to decreased Treg cell numbers and abnormal function. |
| keywords:Myasthenia gravis miR-125a-5p Regulatory T cell |
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