振保丸通过诱导 HSP27 表达减少 Treg 细胞的百分比。

Zhenbao Pill reduces the percentage of Treg cells by inducing HSP27 expression.

机构信息

Department of Orthopaedic Surgery, Renmin Hospital of Wuhan University, Wuhan, 430060 Hubei, China.

Department of Spine Surgery, Inner Mongolia People's Hospital, Hohhot, 010017, Inner Mongolia, China.

出版信息

Biomed Pharmacother. 2017 Dec;96:818-824. doi: 10.1016/j.biopha.2017.09.133. Epub 2017 Nov 6.

DOI:10.1016/j.biopha.2017.09.133

PMID:29078259

Abstract

CONTEXT

Zhenbao pill containing Nacre, Safflower, Musk and Cornu Bubahas has been proved to have a good therapeutic effect on the repair of spinal cord injury (SCI). However, its complex mechanism of repairing SCI is not yet known.

OBJECTIVE

This study attempts to investigate the role of CD4+CD25+Foxp3+ regulatory T cells (Tregs) in the mechanism of the action of Zhenbao pill, and further explore the relationship between Tregs and HSP27 expression in repair mechanism.

MATERIALS AND METHODS

Treatment of human peripheral blood mononuclear cells (PBMCs) with different concentrations (0, 0.5, 2.5, 5, 10mg/mL) of Zhenbao pill, flow cytometry was used to detect the expression of the specific factors CD4+CD25+Foxp3+ in Tregs, and detection of Tregs related regulatory factor TGF-β content was performed with ELISA assay. The relationship between miR-214 and HSP27 was assessed by Luciferase assay, and the level of miR-214 was detected by qPCR. The expression of HSP27 was examined with qPCR and western blotting. RNA interference technology and gene recombination were used to inhibit and up-regulate the expression of HSP27.

RESULTS

Zhenbao pill with 11.61mg/mL of IC for Tregs can significantly inhibit the differentiation into Tregs in human PBMCs and up-regulate by more than 1-fold of HSP27 expression. Essentially, it enhanced the expression of HSP27 by inhibiting miR-214 expression (50%). Inhibition of HSP27 expression, followed by the differentiation into Tregs, was promoted in human PBMCs. When the HSP27 expression was up-regulated, the differentiation into Tregs was decreased by 30%. It indicated that the expression of HSP27 regulated the differentiation into Tregs. Inhibition of HSP27 expression and Zhenbao pill treatment, the differentiation into Tregs was decreased but remained at a higher level than that of the group was only treated with pill. Under the action of Zhenbao pill, the expression of HSP27 was not completely interfered, and its expression level was still increased.

CONCLUSIONS

In the process of repairing the SCI, Zhenbao pill inhibits reduces numbers of Treg lymphocytes as well as TGF-β levels by inducing HSP27 expression.

摘要

背景

珍珠宝胶囊含有珍珠、红花、麝香和鹿茸，已被证明对脊髓损伤（SCI）的修复有很好的治疗效果。然而，其修复 SCI 的复杂机制尚不清楚。

目的

本研究试图探讨 CD4+CD25+Foxp3+调节性 T 细胞（Tregs）在珍珠宝胶囊作用机制中的作用，并进一步探讨 Tregs 与 HSP27 表达在修复机制中的关系。

材料和方法

用不同浓度（0、0.5、2.5、5、10mg/ml）的珍珠宝胶囊处理人外周血单个核细胞（PBMCs），流式细胞术检测 Tregs 中特异性因子 CD4+CD25+Foxp3+的表达，酶联免疫吸附试验（ELISA）检测 Tregs 相关调节因子 TGF-β的含量。用荧光素酶测定法评估 miR-214 与 HSP27 的关系，用 qPCR 检测 miR-214 的水平。用 qPCR 和 Western blot 检测 HSP27 的表达。采用 RNA 干扰技术和基因重组技术抑制和上调 HSP27 的表达。

结果

珍珠宝胶囊对 Tregs 的 IC 为 11.61mg/ml，能明显抑制人 PBMCs 向 Tregs 分化，并使 HSP27 表达上调 1 倍以上。实质上，通过抑制 miR-214 的表达（50%）增强 HSP27 的表达。抑制 HSP27 的表达，促进人 PBMCs 向 Tregs 分化。当 HSP27 表达上调时，Tregs 的分化减少了 30%。表明 HSP27 的表达调节 Tregs 的分化。抑制 HSP27 的表达和珍珠宝胶囊处理后，Tregs 的分化减少，但仍保持在较高水平，高于仅用药物处理的组。在珍珠宝胶囊的作用下，HSP27 的表达并未完全受到干扰，其表达水平仍有所升高。