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[虫草提取物对哮喘缓解期儿童外周血单个核细胞细胞因子及转录因子的影响]

[Effects of Cordyceps extract on cytokines and transcription factors in peripheral blood mononuclear cells of asthmatic children during remission stage].

作者信息

Sun Wen, Yu Jian, Shi Yu-min, Zhang Hao, Wang Ying, Wu Bing-bing

机构信息

Department of Traditional Chinese Medicine, Children's Hospital, Fudan University, Shanghai 201102, China.

出版信息

Zhong Xi Yi Jie He Xue Bao. 2010 Apr;8(4):341-6. doi: 10.3736/jcim20100407.

Abstract

OBJECTIVE

To explore the effects of Cordyceps extract in regulating the imbalance of Th1/Th2 ratio and inhibiting the inflammatory reaction, and to find the theoretical basis of Cordyceps extract for treating asthma in remission stage.

METHODS

A total of 20 peripheral venous blood samples (3 mL) were collected from 20 asthmatic children during remission stage, and peripheral blood mononuclear cells (PBMCs) were separated by Ficoll method. PBMCs were separated into three groups (blank group, low-dose group and high-dose group). The PBMCs were incubated in vitro for 48 hours in the absence (blank group) or presence (low-dose group and high-dose group) of Cordyceps extract at different concentrations (10, 20 microg/mL). The expressions of interferon-gamma (IFN-gamma), interleukin-4 (IL-4), IL-10, T-box expressed in T cells (T-bet), GATA-binding protein-3 (GATA-3) and forkhead/winged-helix transcription factor-3 (Foxp3) mRNAs in PBMCs were measured by real-time fluorescent quantitative polymerase chain reaction, and the contents of IFN-gamma, IL-4 and IL-10 in supernatants were measured by enzyme-linked immunosorbent assay.

RESULTS

The expressions of IFN-gamma mRNA showed no significant differences among the three groups. The expressions of the IL-4 mRNA in the high-dose group and the low-dose group were lower than that in the blank group (P=0.014, P=0.011). The expression of IL-10 mRNA in the high-dose group was higher than that in the blank group (P=0.034). And the differences of the IFN-gamma mRNA/IL-4 mRNA ratio presented no statistic significance among the three groups. The level of IL-4 content in the high-dose group was lower than that in the blank group (P=0.018), but the level of IL-10 content, and ratio of IFN-gamma/IL-4 in the high-dose group were higher than those in the blank group (P=0.011, P=0.045). The differences of the IFN-gamma presented no statistic significance among the three groups. The T-bet/GATA-3 ratio and Foxp3 mRNA expression in the high-dose group were higher than those in the blank group (P=0.001, P=0.015). There was significant difference in expression of GATA-3 mRNA between the high-dose group and the blank group (P=0.028), and between the low-dose group and the blank group (P=0.019). The expression differences of T-bet mRNA were insignificant between any two groups.

CONCLUSION

Cordyceps extract can inhibit the proliferation and differentiation of Th2 cells and reduce the expression of related cytokines by down-regulating the expression of GATA-3 mRNA and up-regulating the expression of Foxp3 mRNA in PBMCs. Meanwhile, it can alleviate the chronic allergic inflammation by increasing the content of IL-10.

摘要

目的

探讨虫草提取物对调节Th1/Th2比例失衡及抑制炎症反应的作用,为虫草提取物治疗哮喘缓解期缓解期哮喘寻找理论依据。

方法

采集20例缓解期哮喘患儿外周静脉血标本(3 mL),采用Ficoll法分离外周血单个核细胞(PBMCs)。将PBMCs分为三组(空白组、低剂量组和高剂量组)。PBMCs在无虫草提取物(空白组)或不同浓度(10、20 μg/mL)的虫草提取物存在下(低剂量组和高剂量组)体外培养48小时。采用实时荧光定量聚合酶链反应检测PBMCs中干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)、IL-10、T细胞特异性转录因子(T-bet)、GATA结合蛋白3(GATA-3)和叉头/翼状螺旋转录因子3(Foxp3)mRNA的表达,采用酶联免疫吸附测定法检测上清液中IFN-γ、IL-4和IL-10的含量。

结果

三组间IFN-γ mRNA表达无显著差异。高剂量组和低剂量组IL-4 mRNA表达低于空白组(P = 0.014,P = 0.011)。高剂量组IL-10 mRNA表达高于空白组(P = 0.034)。三组间IFN-γ mRNA/IL-4 mRNA比值差异无统计学意义。高剂量组IL-4含量低于空白组(P = 0.018),但高剂量组IL-10含量及IFN-γ/IL-4比值高于空白组(P = 0.011,P = 0.045)。三组间IFN-γ差异无统计学意义。高剂量组T-bet/GATA-3比值和Foxp3 mRNA表达高于空白组(P = 0.001,P = 0.015)。高剂量组与空白组GATA-3 mRNA表达差异有统计学意义(P = 0.028),低剂量组与空白组差异有统计学意义(P = 0.019)。任意两组间T-bet mRNA表达差异无统计学意义。

结论

虫草提取物可通过下调PBMCs中GATA-3 mRNA表达、上调Foxp3 mRNA表达,抑制Th2细胞增殖分化,降低相关细胞因子表达。同时,可通过增加IL-10含量减轻慢性过敏性炎症。

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