PcDNA3.0/p27^(kip1)真核表达质粒对小鼠哮喘模型Th1/Th2的影响  

Proportional influence of Th1/Th2 lymphocytes after transferring the recombinant of PcDNA3.0/p27^(kip1) eukaryotic expressing plasmid into asthma model of mice

作　　者：孙洋[1] 陈萍[2] 赵海涛[2] 孙丽[2] 

机构地区：[1]第四军医大学西京医院呼吸内科,西安710032 [2]沈阳军区总医院呼吸内科,沈阳110016

出　　处：《中国免疫学杂志》2010年第8期736-739,共4页Chinese Journal of Immunology

基　　金：军队医药卫生"十一五"科研项目(06MA050)

摘　　要：目的:探讨细胞周期负性调控蛋白PcDNA3.0/p27kip1重组真核表达质粒转染小鼠哮喘模型对Th1/Th2比例失衡的调节作用。方法:小鼠各10只分为A组(正常对照组)、B组(哮喘模型组)、C组(质粒治疗组)三组。用卵蛋白致敏法构建小鼠哮喘模型,以本实验室构建的PcDNA3.0/p27kip1重组真核表达质粒经静脉转染小鼠哮喘模型,36小时后Western blot检测肺组织中p27蛋白表达量;流式细胞仪分析小鼠脾CD4+、CD8+T细胞比例;ELISA检测小鼠血清IL-4、IL-5、IL-13、IFN-γ水平。结果:B组与A组比较,哮喘发病后肺p27蛋白表达量明显降低;脾CD4+T细胞显著增高而CD8+T细胞显著降低;血清IL-4、IL-5、IL-13水平明显升高而IFN-γ明显降低。C组与B组比较,肺p27蛋白表达量明显增加,脾CD4+T细胞比例显著降低而CD8+T细胞显著升高;血清IL-4、IL-5、IL-13水平明显降低而IFN-γ明显升高。结论:PcDNA3.0/p27kip1静脉给药可在哮喘模型肺组织中表达,并通过基因干预调控细胞周期的方式调节Th1/Th2比例失衡,为哮喘防治提出了新思路。Objective：To investigate the effects of the proportional imbalance of Th1/Th2 in allergic asthma model by transferring the recombinant of PcDNA3.0/p27^kip1 eukaryotic expressing plasmid into allergic asthma model.Methods：KM mice were distributed into three groups：healthy control mice（group A）;Ovalbumin（OVA）-sensitized/challenged mice（group B） and OVA-sensitized/challenged mice intratracheally administered with PcDNA3.0/p27^kip1 plasmid（plasmid-treated mice,group C）.KM mice were sensitized and challenged intratracheally with OVA,then transfected with the recombinant PcDNA3.0/p27^kip1 eukaryotic expressing plasmid which was constructed by our laboratory crew through vein injection.The p27 protein expression in lungs 36 h after transfection was analyzed by Western blot;Flow cytometry was used to analyze the proportion of CD4 （＋）and CD8（＋） T lymphocyte subsets in the spleen.The serum levels of IL-4,IL-5,IL-13 and IFN-γ were measured by ELISA.Results：Compared to the group A,the expression of p27 protein was significantly reduced,and the proportion of CD4（＋）lymphocytes and serum levels of IL-4,IL-5 and IL-13 were significantly higher but CD8（＋）lymphocytes and IFN-γ was significantly decreased in group B.However,in group C,the expression of p27 protein was significantly increased;Whereas the proportion of CD4 （＋） lymphocytes,serum levels of IL-4,IL-5 and IL-13 were significantly decreased.CD8 （＋） lymphocytes and IFN-γ were significantly increased than those in group B.Conclusion：p27 could be expressed in lungs of plasmid-treated mice and regulate the proportion of Th1/Th2 subset by means of adjusting cell cycles.This method may serve as a potential therapeutic option for the treatment of asthma.

关 键 词：P27 基因治疗 免疫调节 哮喘 

分 类 号：R562.2[医药卫生—呼吸系统]