上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (7): 962-.doi: 10.3969/j.issn.1674-8115.2011.07.020

• 论著（临床研究） • 上一篇    下一篇

不同刺激物对过敏性哮喘产妇脐血中嗜碱性粒细胞分泌IL-4的影响

杨 玲1, 许以平2, 姚苏杭2, 熊 瑛3, 祝 捷4, 王克敏5   

  1. 1．上海交通大学 医学院附属新华医院老年科, 上海 200092； 2.上海交通大学 |医学院附属仁济医院过敏中心, 上海 200127； 3.上海交通大学 医学院附属新华医院妇产科, 上海 200092； 4.上海交通大学 |医学院附属仁济医院妇产科, 上海 200127； 5.上海交通大学 基础医学院生物化学与分子生物学教研室, 上海 200025
  • 出版日期:2011-07-28 发布日期:2011-07-27
  • 作者简介:杨 玲（1973—）, 女, 副主任医师, 博士；电子信箱: yanglingjoy@hotmail.com。
  • 基金资助:

    上海交通大学医工合作课题（YG2010MS12）

Effects of various stimuli on IL-4 release of umbilical cord blood-derived basophils from neonates of mothers with allergic asthma

YANG Ling1, XU Yi-ping2, YAO Su-hang2, XIONG Ying3, ZHU Jie4, WANG Ke-min5   

  1. 1.Department of Geriatrics, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China；2.Allergy Center, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China；3.Department of Obstetrics and Gynecology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China；4.Department of Obstetrics and Gynecology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200127, China；5.Department of Biochemistry and Molecular Biology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2011-07-28 Published:2011-07-27
  • Supported by:

    Shanghai Jiaotong University Foundation, YG2010MS12

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摘要：

目的 研究不同刺激物干预对过敏性哮喘产妇新生儿脐血中嗜碱性粒细胞分泌白介素4（IL-4）的影响。方法 将源自过敏性哮喘产妇和健康产妇的新生儿脐血分为哮喘组（n=12）和正常对照组（n=16），采用流式细胞术分选提纯嗜碱性粒细胞，锥虫蓝染色鉴定细胞活性。在两组提纯后的嗜碱性粒细胞中分别加入ACM缓冲液（阴性对照）、肽聚糖(PGN)、粉尘螨浸液及甘露醇，采用ELISA法测定细胞培养上清液中IL-4的质量浓度。结果 细胞分选结果显示：嗜碱性粒细胞纯度为（95.64±3.15）%，回收率为（60.22±7.18）%；锥虫蓝染色显示细胞活性为（99.4±0.89）%。与阴性对照比较，经PGN、粉尘螨浸液和甘露醇刺激后的哮喘组细胞培养上清液中IL-4质量浓度显著升高（P<0.05或P<0.01）；经PGN和甘露醇刺激后，哮喘组细胞培养上清液中IL-4质量浓度显著高于正常对照组（P<0.05和P<0.01）。结论 PGN、粉尘螨及甘露醇可刺激过敏性哮喘产妇新生儿脐血中嗜碱性粒细胞IL-4的分泌增加，通过诱导辅助性T淋巴细胞亚群Th2细胞的分化，参与哮喘的发生和发展。

关键词: 过敏性哮喘, 嗜碱性粒细胞, 脐血, 白介素4

Abstract:

Objective To investigate the effects of various stimuli on interleukin-4 (IL-4) release of umbilical cord blood-derived basophils from neonates of mothers with allergic asthma. Methods Samples of umbilical cord blood from neonates of mothers with allergic asthma and healthy mothers were divided into asthma group （n=12） and normal control group （n=16）. Basophils were sorted and purified by flow cytometry, and the cell viability was identified by trypan blue staining. The purified basophils were incubated with ACM buffer (negative control), peptidoglycan (PGN), Dermatophagoides farinae and mannitol respectively, and the mass concentrations of IL-4 in the supernatants of culture fluid were detected by ELISA. Results The result of cell sorting indicated that the purity of basophils was (95.64±3.15)%, with the recovery rate of (60.22±7.18)%, and trypan blue staining revealed that the cell viability was (99.4±0.89)%. Compared with negative control, the mass concentrations of IL-4 in the supernatants of culture fluid significantly increased after stimulation by PGN, Dermatophagoides farinae and mannitol in asthma group （P<0.05 or P<0.01）. After stimulation by PGN and mannitol, the mass concentrations of IL-4 in the supernatants of culture fluid in asthma group were significantly higher than those in normal control group （P<0.05 and P<0.01）. Conclusion PGN, Dermatophagoides farinae and mannitol may stimulate the IL-4 release of umbilical cord blood-derived basophils from neonates of mothers with allergic asthma, induce differentiation of Th2 cells and participate in the pathogenesis of asthma.

Key words: allergic asthma, basophil, umbilical cord blood, interleukin-4