| 贾宏林,张 茹,梁晓鹰,杨 清,姜孝芳.5-氮杂胞苷对T淋巴细胞Jurkat miR-126及IFN-γ、GATA3、ROR-γ、Foxp3表达的影响[J].现代生物医学进展英文版,2022,(1):43-51. |
| 5-氮杂胞苷对T淋巴细胞Jurkat miR-126及IFN-γ、GATA3、ROR-γ、Foxp3表达的影响 |
| Effects of 5-azacytidine on Expression of Jurkat miR-126 and IFN-γ, GATA3, ROR-γ and Foxp3 in T Lymphocytes |
| Received:July 23, 2021 Revised:August 18, 2021 |
| DOI：10.13241/j.cnki.pmb.2022.01.007 |
| 中文关键词: 变应性鼻炎 T淋巴细胞 DNA甲基化 5-氮杂胞苷 miR-126 Th细胞 |
| 英文关键词: Allergic rhinitis T lymphocytes DNA methylation 5-aza miR-126 Th cell |
| 基金项目:国家自然科学基金项目（81760182） |
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| 中文摘要: |
| 摘要 目的：探究甲基化酶抑制剂5-氮杂胞苷（5-azacytidin, 5-aza）对T淋巴细胞（Jurkat）miR-126、Th1/Th2、Th17/Treg细胞亚群及因子IFN-γ、GATA3、ROR-γ和Foxp3的调控作用。方法：采用不同浓度5-aza干预T淋巴细胞，24 h、48 h后检测其对细胞增殖抑制作用；实时荧光定量PCR、Western Blot检测5-aza干预后miR-126表达水平以及IFN-γ、GATA3、ROR-γ和Foxp3的mRNA及蛋白表达水平；流式细胞术检测5-aza干预后Th1/Th2、Th17/Treg细胞亚群分化比例。结果：甲基化酶抑制剂干预T淋巴细胞后，细胞抑制率随5-aza浓度增大及作用时间延长呈递增趋势（P<0.01）；细胞抑制率在24 h、48 h，低、中、高浓度下分别为（14.73±0.93）%、（32.67±8.40）%、（60.87±5.78）%以及(18.98±0.73)%、（39.80±8.42）%、（64.11±6.04）%；抑制率在24 h与48 h之间无差异（P>0.05）。甲基化酶抑制剂干预后miR-126表达降低（P<0.01）； IFN-γ蛋白表达降低（P<0.05）、Th1细胞亚群数目降低（P<0.01）； GATA3 mRNA和蛋白表达升高（P<0.05）、Th2细胞亚群数目增加（P<0.01）；ROR-γ蛋白表达降低（P>0.05）、Th17细胞亚群数目降低（P<0.05）；Foxp3 mRNA和蛋白表达升高（P>0.05）、Treg细胞亚群数目增加（P<0.05）。结论：甲基化酶抑制剂可以下调Jurkat细胞miR-126基因表达；下调Th1 、Th17细胞亚群分化，上调Th2、Treg细胞亚群分化；调控细胞因子IFN-γ、GATA3、ROR-γ和Foxp3的表达。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the regulatory effects of methylase inhibitor 5-azacytidin (5-aza) on miR-126, Th1/Th2, Th17/Treg cell subsets and cytokines IFN-γ, GATA3, ROR-γ and Foxp3 in T lymphocytes (Jurkat). Methods: T lymphocytes were treated with different concentrations of 5-aza. The inhibitory effects of 5-aza on T lymphocytes proliferation were measured at 24 h and 48 h after treatment. Real-time quantitative PCR and Western Blot were used to detect the expression level of miR-126 and the mRNA and protein expression levels of IFN-γ, GATA3, ROR-γ and Foxp3 after 5-aza intervention; Flow cytometry was used to detect the differentiation ratio of Th1/Th2 and Th17/Treg cell subsets after 5-aza treatment. Results: The cytostatic rate increased with increasing 5-aza concentration and duration of action after methylation enzyme inhibitor intervention on T lymphocytes (P<0.01). The cell inhibitory rates were (14.73±0.93)%, (32.67±8.40)%, (60.87±5.78)% and (18.98±0.73)%, (39.80±8.42)%, (64.11±6.04)% at 24 h, 48 h, low, medium and high concentrations, respectively. There was no difference in inhibition between 24 h and 48 h (P>0.05). The expression of miR-126 decreased after methylase inhibitor intervention (P<0.01); The expression of IFN-γ protein decreased (P<0.05), the number of Th1 cell subsets decreased (P<0.01); GATA3 mRNA and protein expression increased (P<0.05), the number of Th2 cell subsets increased (P<0.01); The expression of ROR-γ protein was decreased (P>0.05), the number of Th17 cell subsets decreased (P<0.05); Foxp3 mRNA and protein expression increased (P>0.05), the number of Treg cell subsets increased (P<0.05). Conclusion: Methylase inhibitors could down-regulate miR-126 gene expression in Jurkat cells. The down-regulation of subpopulation differentiation of Th1 and Th17 cells, up-regulation of subpopulation differentiation of Th2 and Treg cells, and expression of cytokines IFN-γ, GATA3, ROR-γ and Foxp3 were regulated by methylase inhibitors. |
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