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[香烟烟雾暴露对哮喘大鼠CD4⁺CD25⁺调节性T细胞百分比及转录因子Foxp3表达的影响]

[Effect of cigarette smoke exposure on the percentage of CD(4)(+)CD(25)(+) regulatory T cells and the expression of transcription factor Foxp3 in asthmatic rats].

作者信息

Jiang Yi, DU Yong-Cheng, Xu Jian-Ying

机构信息

Department of Respiration Medicine, the First Hospital of Shanxi Medical University, Taiyuan 030001, China.

出版信息

Zhonghua Jie He He Hu Xi Za Zhi. 2010 Aug;33(8):582-6.

Abstract

OBJECTIVE

To investigate the effect of cigarette smoke exposure on the ratio of CD(4)(+)CD(25)(+) regulatory T cells (Treg) and expression of transcription factor Foxp3 in asthmatic rats.

METHODS

Forty male Wistar rats were randomly divided into 4 groups (n = 10 for each group): a normal saline group, an aerosolized ovalbumin (OVA) exposure group, a cigarette smoke exposure group and a combined OVA and cigarette smoke exposure group. The rats were exposed to air or cigarette smoke and to normal saline or OVA aerosol for 8 weeks respectively. The percentage of CD(4)(+)CD(25)(+) T cells was determined by flow cytometry analysis. The concentration of interleukin-4 (IL-4) and interferon-γ (INF-γ) in peripheral blood and lung homogenates were measured by enzyme-linked immunosorbent assay (ELISA). The protein expression of Foxp3 in the lung was detected by Western blot.

RESULTS

(1) The percentage of CD(4)(+)CD(25)(+) T cells in aerosolized OVA group [(6.4 ± 1.0)%] was significantly lower than that in the normal saline group [(9.9 ± 1.0)%] (P < 0.01). The percentage of CD(4)(+)CD(25)(+) T cells [(3.3 ± 0.8)%] in OVA combined cigarette smoke exposure group was remarkably lower than that in the aerosolized OVA exposure group and in the normal saline group(P < 0.01). (2) IL-4 in both plasma and lung [(22.6 ± 4.3) ng/L, (0.8 ± 0.1) ng/L] was significantly increased in the OVA-exposed rats compared with the normal saline group [(11.4 ± 2.9) ng/L, (0.3 ± 0.1) ng/L] (P < 0.01). Further remarkable increase in IL-4 of both plasma and lung was observed in the group exposed to both OVA and cigarette smoke [(34.1 ± 6.1) ng/L, (1.4 ± 0.3) ng/L] compared with the aerosolized OVA exposure group and the normal saline group (P < 0.05). INF-γ of plasma in OVA-exposed group [(59 ± 20) ng/L] was significantly decreased compared with the normal saline group [(151 ± 56) ng/L] (P < 0.01), and a further remarkable decrease in INF-γ of plasma was observed in the group exposed to both OVA and cigarette smoke [(10 ± 3) ng/L] compared with the aerosolized OVA exposure group and the normal saline group (P < 0.05). (3) Protein expression of Foxp3 in the aerosolized OVA group (8.18 ± 0.26) was lower than that in the normal saline group (10.27 ± 0.33, P < 0.01), while the protein expression of Foxp3 in OVA combined cigarette smoke exposure group (6.36 ± 0.38) was lower than that in the normal saline group and the aerosolized OVA exposure group (P < 0.01).

CONCLUSION

The number of CD(4)(+)CD(25)(+) Treg cells and the expression of Foxp3 are likely to be altered by cigarette smoke exposure, which might play an important role in cigarette smoking-induced Th1/Th2 imbalance in asthmatic rats.

摘要

目的

探讨香烟烟雾暴露对哮喘大鼠CD4⁺CD25⁺调节性T细胞(Treg)比例及转录因子Foxp3表达的影响。

方法

40只雄性Wistar大鼠随机分为4组(每组n = 10):生理盐水组、雾化卵清蛋白(OVA)暴露组、香烟烟雾暴露组和OVA与香烟烟雾联合暴露组。大鼠分别暴露于空气或香烟烟雾以及生理盐水或OVA气雾剂中8周。通过流式细胞术分析测定CD4⁺CD25⁺T细胞的百分比。采用酶联免疫吸附测定(ELISA)法检测外周血和肺匀浆中白细胞介素-4(IL-4)和干扰素-γ(INF-γ)的浓度。通过蛋白质印迹法检测肺组织中Foxp3的蛋白表达。

结果

(1)雾化OVA组CD4⁺CD25⁺T细胞百分比[(6.4±1.0)%]显著低于生理盐水组[(9.9±1.0)%](P < 0.01)。OVA与香烟烟雾联合暴露组CD4⁺CD25⁺T细胞百分比[(3.3±0.8)%]显著低于雾化OVA暴露组和生理盐水组(P < 0.01)。(2)与生理盐水组[(11.4±2.9)ng/L,(0.3±0.1)ng/L]相比,OVA暴露大鼠血浆和肺组织中的IL-4[(22.6±4.3)ng/L,(0.8±0.1)ng/L]显著升高(P < 0.01)。与雾化OVA暴露组和生理盐水组相比,OVA与香烟烟雾联合暴露组血浆和肺组织中的IL-4进一步显著升高[(34.1±6.1)ng/L,(1.4±0.3)ng/L](P < 0.05)。OVA暴露组血浆中的INF-γ[(59±20)ng/L]与生理盐水组[(151±56)ng/L]相比显著降低(P < 0.01),与雾化OVA暴露组和生理盐水组相比,OVA与香烟烟雾联合暴露组血浆中的INF-γ进一步显著降低[(10±3)ng/L](P < 0.05)。(3)雾化OVA组Foxp3的蛋白表达(8.18±0.26)低于生理盐水组(10.27±0.33,P < 0.01),而OVA与香烟烟雾联合暴露组Foxp3的蛋白表达(6.36±0.38)低于生理盐水组和雾化OVA暴露组(P < 0.01)。

结论

香烟烟雾暴露可能会改变CD4⁺CD25⁺Treg细胞数量及Foxp3表达,这可能在吸烟诱导的哮喘大鼠Th1/Th2失衡中起重要作用。

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