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1型糖尿病患者血清miRNA-24、mTOR与Treg细胞亚群比例的相关性

Correlation between serum miRNA-24, mTOR and ratio of Treg cell subsets in patients with type 1 diabetes mellitus

    摘要
  • 摘要:
    目的 探讨1型糖尿病（T1DM）患者血清miRNA-24、哺乳动物雷帕霉素靶蛋白（mTOR）与外周血单个核细胞（PBMC）中调节性T细胞（Treg）及其亚群比例的相关性。
    方法 选择2020年1月至2022年1月在南通大学附属常熟医院内分泌科就诊的40例T1DM患者及同期体检健康者40例（对照组）。用密度梯度离心法分离研究对象的PBMC，以CD25和CD127标记CD4+ Treg（CD4+CD25hiCD127low，总Treg）；分别以CD45RA、CD45RO标记效应型Treg细胞亚群（CD45RA+Treg，eTreg）、静息型Treg细胞亚群（CD45RO+Treg，rTreg）。采用qPCR法检测血清miRNA-24的表达，ELISA检测血清mTOR的水平，比较两组上述指标水平。采用Pearson法分析miRNA-24及mTOR与T1DM患者的eTreg、rTreg及eTreg/rTreg比值之间的相关性。
    结果 两组PBMC中总Treg比例差异无统计学意义。T1DM组eTreg亚群占比低于对照组、rTreg亚群占比高于对照组（P＜0.05）；T1DM组eTreg/rTreg比值小于对照组（P＜0.05）。T1DM组患者血清中miRNA-24水平低于对照组（P＜0.001），mTOR水平高于对照组（P＜0.05）。T1DM患者单糖尿病抗体亚组（n＝23）与多糖尿病抗体亚组（n＝17）总Treg细胞比例、Treg亚群比例、eTreg/rTreg比值、miRNA-24及mTOR水平差异均无统计学意义；血糖控制良好（糖化血红蛋白＜7.5%）亚组（n＝11）与血糖控制不佳（糖化血红蛋白≥7.5%）亚组（n＝29）上述指标差异均有统计学意义（均P＜0.05）。miRNA-24与eTreg（r＝0. 40，P＝0. 01）、eTreg/rTreg比值（r＝0.66，P＜0.001）正相关，与rTreg为负相关（r＝-0. 69，P＜0.001）；mTOR与eTreg（r＝-0. 32，P＝0. 04）、eTreg/rTreg比值（r＝-0.40，P＝0.01）负相关，与rTreg正相关（r＝0. 48，P＝0.001）。
    结论 T1DM患者PBMC中CD4+Treg亚群改变，不同血糖控制T1DM患者miRNA-24、mTOR水平不同，miRNA-24、mTOR与Treg亚群比例及eTreg/rTreg比值存在一定相关性，提示miRNA-24、mTOR可能参与T1DM患者Treg调节。

     

    Abstract:
    Objective To explore the correlation between serum miRNA-24, mechanism target of rapamycin (mTOR) and the frequency of regulatory T cells (Treg) and their subsets in peripheral blood mononuclear cells (PBMC) in patients with type 1 diabetes (T1DM).
    Methods A total of 40 T1DM patients who were treated in the Endocrinology Department of Affiliated Changshu Hospital of Nantong University from January 2020 to January 2022 were selected as T1DM group, and 40 healthy people who underwent physical examination in the hospital during the same period were selected as the control group. CD4+Treg (CD4+CD25hiCD127low) were marked with CD25 and CD127, and Treg cell subsets included effector Treg (eTreg, CD45RA+Treg) and resting Treg (rTreg, CD45RO+Treg) were differentiated by CD45RA and CD45RO. Serum miRNA-24 was detected by qPCR and serum mTOR was detected by enzyme-linked immunosorbent assay. The levels of above indexes were compared between the two groups. Pearson test was used to analyze the correlations between miRNA-24 and mTOR levels and eTreg, rTreg and eTreg/rTreg ratio in T1DM patients.
    Results There was no statistically significant difference in the total Treg ratio in the PBMC between T1DM group and control group. Compared with the control group, the proportion of eTreg subset in the T1DM group was lower, the proportion of rTreg subset was higher (P < 0.05), and the eTreg/rTreg ratio in the T1DM group was lower (P < 0.05). Compared with the control group, the serum miRNA-24 level in the T1DM group was lower (P < 0.001), and the mTOR level was higher (P < 0.05). There was no significant difference in the proportion of total Treg cells, the proportion of Treg subsets, eTreg/rTreg ratio, the levels of miRNA-24 and mTOR between the single-antibody subgroup (n＝23) and multi-antibody subgroup (n＝17) among T1DM patients. There were statistically difference in the above indicators between the blood glucose controlled well (glycosylated hemoglobin < 7.5%, n＝11) and bad (glycosylated hemoglobin≥7.5%, n＝29) subgroups among T1DM patients (P < 0.05). MiRNA-24 was positively correlated with eTreg (r＝0.40, P＝0.01) and eTreg/rTreg ratio (r＝0.66, P < 0.001), while was negatively correlated with rTreg (r＝-0.69, P < 0.001); mTOR was negatively correlated with eTreg (r＝-0.32, P＝0.04) and eTreg/rTreg ratio (r＝-0.40, P＝0.01), while was positively correlated with rTreg (r＝0.48, P＝0.001).
    Conclusion CD4+Treg subsets in PBMC in patients with T1DM change; miRNA-24 and mTOR might be different. MiRNA-24 and mTOR might be correlated with the frequency of Treg cell subsets and eTreg/rTreg ratio, and involve in the regulation of Treg in T1DM patients.

     

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