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特发性血小板减少性紫癜患者CD4+CD25+CD127low调节性T细胞、转化生长因子-β及Notch1 mRNA的表达及意义

[Expression and Significance of CD4+ CD25+ CDl27low Regulatory T Cells, TGF-β and Notch1 mRNA in Patients with Idiopathic Thrombocytopenic Purpura].

作者信息

Huang Wen-Ye, Sun Qiu-Hui, Chen Ye-Ping

机构信息

Department of Hematology, Yueqing Hospital Affiliated to Wenzhou Medical University, Wenzhou 325600, Zhejiang Province, China. E-mail:

Department of Hematology, Yueqing Hospital Affiliated to Wenzhou Medical University, Wenzhou 325600, Zhejiang Province, China.

出版信息

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2015 Dec;23(6):1652-6. doi: 10.7534/j.issn.1009-2137.2015.06.023.

Abstract

OBJECTIVE

To detect the expression levels of CD4(+) CD25(+) CDl27(low) Treg cells, TGF-β and Notch1 mRNA in peripheral blood of the patients with idiopathic thrombocytopenic purpura (IPT) before and after treatment, and to investigate their significance in the pathogenesis of ITP.

METHODS

Peripheral blood was collected from 30 newly diagnosed patients with ITP and 20 normal controls, then the number of CD4(+) CD25(+) Treg and CD4(+) CD25(+) CDl27(low) Treg were detected by the flow cytometry. Plasma TGF-β level was determined by ELISA. Total RNA was extracted and the expression level of Notch1 mRNA was measured by real-time Q-PCR.

RESULTS

The expression levels of CD4(+) CD25(+) CDl27(low) Treg and CD4(+) CD25(+) Treg in newly diagnosed ITP group were significantly lower than those in normal controls. After treatment, the proportion of Tregs increased to (5.17% ± 0.74%) and (4.16% ± 0.68%), and was higher than that in newly diagnosed patients. The TGF-β level in peripheral blood of newly-diagnosed patients was obviously lower than that in normal controls, and was (961.53 ± 60.10) ng/L after treatment and was significantly higher than that in newly diagnosed patients; the expression level of Notch1 mRNA in peripheral blood of patients in newly-diagnosed group was obviously lower than that in control, and was (1.35 ± 0.10) after treatment that was higher than that in newly-diagnosed group. After treatment, the proportion of Treg cells, level of TGF-β and erpression level of Notch1 mRNA in effective group were higher than those in effective group, improved group and ineffective group, and there was significant difference (P <0.01). The expression level of TGF-β and Notch1 mRNA in ITP patients possitively correlated to CD4(+) CD25(+) CDl27(low) (P <0.01).

CONCLUSIONS

The levels of CD4(+) CD25(+) CDl27(low) Treg, TGF-βand Notch1 mRNA in peripheral blood of the patients with ITP are significantly lower than those of normal control, suggesting that there is significant abnormal immunoregulation in ITP patients. In the ITP patients the levels of CD4(+) CD25(+) CDl27(low) Treg postively correlated with Notch1 mRNA expression, indicating that Notch signal may be revalent to Treg's immunosuppression function.

摘要

目的

检测特发性血小板减少性紫癜(IPT)患者治疗前后外周血中CD4(+)CD25(+)CD127(低)调节性T细胞(Treg)、转化生长因子-β(TGF-β)及Notch1 mRNA的表达水平,探讨其在ITP发病机制中的意义。

方法

采集30例新诊断ITP患者及20例正常对照者外周血,采用流式细胞术检测CD4(+)CD25(+)Treg及CD4(+)CD25(+)CD127(低)Treg细胞数量。采用酶联免疫吸附测定法(ELISA)检测血浆TGF-β水平。提取总RNA,采用实时荧光定量聚合酶链反应(real-time Q-PCR)检测Notch1 mRNA表达水平。

结果

新诊断ITP组CD4(+)CD25(+)CD127(低)Treg及CD4(+)CD25(+)Treg细胞表达水平显著低于正常对照组。治疗后,Treg细胞比例分别升至(5.17%±0.74%)和(4.16%±0.68%),高于新诊断患者。新诊断患者外周血TGF-β水平明显低于正常对照组,治疗后为(961.53±60.10)ng/L,显著高于新诊断患者;新诊断组患者外周血Notch1 mRNA表达水平明显低于对照组,治疗后为(1.35±0.10),高于新诊断组。治疗后,有效组Treg细胞比例、TGF-β水平及Notch1 mRNA表达水平均高于无效组、改善组,差异有统计学意义(P<0.01)。ITP患者TGF-β及Notch1 mRNA表达水平与CD4(+)CD25(+)CD127(低)呈正相关(P<0.01)。

结论

ITP患者外周血中CD4(+)CD25(+)CD127(低)Treg、TGF-β及Notch1 mRNA水平显著低于正常对照,提示ITP患者存在明显的免疫调节异常。ITP患者中CD4(+)CD25(+)CD127(低)Treg水平与Notch1 mRNA表达呈正相关,提示Notch信号可能与Treg的免疫抑制功能有关。

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