皮内注射灭活卡介苗对哮喘大鼠CD4+CD25+Treg细胞数量和CTLA-4 mRNA表达的影响

doi:10.13481/j.1671-587x.20150316

摘要/Abstract

摘要：

目的:探讨皮内注射灭活卡介苗(BCG)后哮喘大鼠CD4⁺CD25⁺Treg细胞数量、细胞毒性T 淋巴细胞抗原4(CTLA-4)的表达及血清转移生长因子β(TGF-β)水平变化,阐明灭活BCG对CD4⁺CD25⁺Treg细胞数量及CTLA-4 mRNA表达的影响。方法:30只健康雄性成年SD大鼠随机分为:对照组、模型组和BCG组,每组10只;除对照组外,模型组和BCG组大鼠采用卵清蛋白(OVA)致敏3周;再激发6周,构建大鼠慢性哮喘模型。BCG组大鼠在致敏前3 d开始皮内注射BCG,共9周。各组大鼠均行气道高反应性检测,检测CD4⁺CD25⁺Treg细胞数量及CTLA-4 mRNA相对表达水平,行肺泡灌洗液(BALF)中细胞总数及嗜酸性粒细胞(EOS)百分比计数,检测各组大鼠血清中TGF-β水平。结果:模型组大鼠BALF中细胞总数及EOS百分比明显高于对照组(P<0.05),BCG组明显低于模型组(P<0.05)。组胺质量浓度从0.32 g·L^-1开始,模型组和BCG组大鼠气道阻力明显高于对照组 (P<0.05);组胺质量浓度从0.64 g·L^-1开始, BCG组大鼠气道阻力明显低于模型组(P<0.05)。模型组大鼠CD4⁺CD25⁺Treg细胞/CD4⁺细胞数量明显低于对照组(P<0.05),BCG组明显高于模型组(P<0.05)。模型组大鼠CTLA-4 mRNA相对表达水平明显低于对照组(P<0.05),BCG组明显高于模型组(P<0.05)。模型组大鼠血清TGF-β水平明显低于对照组(P<0.05),BCG组明显高于模型组(P<0.05)。结论:灭活BCG改善气道炎症和气道阻力作用可能与恢复CD4⁺CD25⁺Treg细胞数量及功能有关联。

关键词: 灭活卡介苗, 调节性T细胞, 气道炎症, 气道阻力, 哮喘, 大鼠, SD

Abstract:

Objective To observe the changes of number of the CD4⁺CD25⁺regulatory T cells, the expression level of CTLA-4 mRNA and the level of serum TGF-β, and to clarify the effect of inactivated Bacillus Calmette-Guerin vaccine(BCG) on the number of the CD4⁺CD25⁺regulatory T cells and CTLA-4 mRNA expression. Methods Thirty Sprague-Dawley (SD) rats were randomly divided into control group, model group and BCG group (n=10).The rats in model group and BCG group were sensitized by injecting intraperitioneally with OVA for 3 weeks;On the 22th day, they were challenged with 2% OVA for 6 weeks to built the chronic rasthma models.The rats in control group were sensitized and challenged with normal saline.The rats in BCG group were intradermally injected with inactivated BCG 3 d before sensitizing lasting 9 weeks.The airway resistances of the rats in various groups were assessed after intubation.The number of CD4⁺CD25⁺regulatory T cells and the expression level of CTLA-4mRNA were detected.The levels of serum TGF-β were detected by ELISA.The number of cells and the percentage of eosinophil(EOS) in bronchial alveolar lavage fluid (BALF) of the rats in various groups were detected. Results Compared with control group, the total number of cells and the percentage of EOS in BALF of the rats in model group were decreased (P<0.05);compared with model group, the total number of cells and the percentage of EOS in BALF of the rats in BCG group were increased(P<0.05).Compared with control group, the airway resistances of the rats in model group was increased when the concentration of histamine was 0.32 g·L^-1;compared with model group, the airway resistance of the rats in BCG group was decreased when the concentration of histamine was 0.64 g·L^-1.Compared with control group, the number of CD4⁺CD25⁺ regulatory T cells, the expression level of CTLA-4mRNA and the level of serum TGF-β of the rats in model group were decreased (P<0.05);compared with model group, CD4⁺CD25⁺ regulatory T cells, the expression level of CTLA-4 mRNA and the level of serum TGF-β of the rats in BCG group were increased (P<0.05). Conclusion The inactivated BCG can improve the airway inflammation and airway hyperresponsiveness in the asthmatic rats, and the mechanisms are correlated with the the number and the function of CD4⁺CD25⁺ regulatory T cells.

Key words: inactivated Bacillus Calmette-Guerin, CD4⁺CD25⁺regulatory T cells, airway inflammation, airway resistance, asthmas, rats,SD

中图分类号:

R562.25

龚萍, 李云, 罗睿, 李红, 谭钰嫔, 龙毅. 皮内注射灭活卡介苗对哮喘大鼠CD4⁺CD25⁺Treg细胞数量和CTLA-4 mRNA表达的影响[J]. 吉林大学学报(医学版), 2015, 41(03): 517-521.

GONG Ping, LI Yun, LUO Rui, LI Hong, TAN Yupin, LONG Yi. Effect of intradermal inactivated Bacillus Calmette-Guérin vaccine on number of CD4⁺CD25⁺Treg cells and expression of CTLA-4 mRNA in asthmatic rats[J]. Journal of Jilin University Medicine Edition, 2015, 41(03): 517-521.

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