目的:研究五虎汤对病毒诱发幼年哮喘大鼠脾细胞悬液T-bet、GATA-3、RORγt、Foxp3 m RNA表达的影响。方法:采用RSV诱导OVA致敏大鼠哮喘模型。造模成功后,随机分成五虎汤高、中、低[4.752 g/(kg·d)、2.376 g/(kg·d)、1.188 g/(...目的:研究五虎汤对病毒诱发幼年哮喘大鼠脾细胞悬液T-bet、GATA-3、RORγt、Foxp3 m RNA表达的影响。方法:采用RSV诱导OVA致敏大鼠哮喘模型。造模成功后,随机分成五虎汤高、中、低[4.752 g/(kg·d)、2.376 g/(kg·d)、1.188 g/(kg·d)]剂量组、地塞米松组[地塞米松片,2.4 mg/(kg·d))]、模型组。干预14 d后,取脾组织,采用RT-PCR法检测脾细胞悬液中T-bet、GATA-3、RORγt、Foxp3 m RNA表达情况。结果:模型组脾组织T-bet、Foxp3m RNA表达低于空白组(P<0.05),而GATA-3、RORγt m RNA表达高于空白对照组(P<0.05)。五虎汤高、中剂量组脾组织GATA-3、RORγt m RNA表达均明显低于模型组(P<0.01或P<0.05),而T-bet、Foxp3 m RNA表达均明显高于模型组(P<0.01或P<0.05)。结论:五虎汤能调控病毒诱发幼年哮喘大鼠脾组织转录因子T-bet、GATA-3、RORγt、Foxp3 m RNA的表达,调控Thl、Th2、Thl7、Treg细胞,重建免疫平衡,抑制哮喘气道炎症形成。展开更多
目的研究分泌性中耳炎(otitis media with effusion,OME)中Th17/调节性T细胞(tregulatory,Treg)及其相关细胞因子、特异性转录因子的变化,深入探索OME免疫学发病机制。方法同批次健康雄性SD大鼠,随机分为对照组(20只,40耳),实验组(20只...目的研究分泌性中耳炎(otitis media with effusion,OME)中Th17/调节性T细胞(tregulatory,Treg)及其相关细胞因子、特异性转录因子的变化,深入探索OME免疫学发病机制。方法同批次健康雄性SD大鼠,随机分为对照组(20只,40耳),实验组(20只,40耳)。实验组大鼠采用卵清蛋白腹腔致敏及鼓膜穿刺耳内致敏2次制成变应原诱发的变态反应相关的OME SD大鼠模型,对照组予以空白对照处理,基于内镜局部积液征、组织学及电镜观察验证建模成功;从免疫细胞、分子和基因三个水平检测、比较OME组及对照组大鼠外周血及中耳局部变化:通过流式细胞技术(flow cytometry,FCM)、酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)和逆转录-聚合酶链反应(reverse transcription-polymerase chain Reaction,RT-PCR)分别检测外周血Th17细胞和CD4^+CD25^+Foxp3^+Treg细胞、相关细胞因子白细胞介素-17(interleukin 17,IL-17)和转化生长因子-β(transforming growth factor beta,TGF-β);中耳局部对应特异性转录因子维甲酸相关孤核受体γt(retinoid-related orphan receptorγt,ROR-γt)、叉状头转录因子3(forkhead box protein 3,Foxp3)m RNA表达水平在OME组及对照组的变化。结果基于内镜、组织学及电镜评估,成功建立免疫相关的OME大鼠模型;外周血TGF-β浓度测定示实验组:95.79±21.60 pg/ml,对照组:72.49±19.75 pg/ml,实验组较对照组显著升高(P=0.005<0.05),差异有统计学意义;外周血IL-17浓度测定实验组与对照组差异无统计学意义(P>0.05);实验组与对照组外周血Th17细胞、CD4^+CD25^+Foxp3^+Treg细胞及两者比值测定差异无统计学意义(P>0.05)。中耳局部检测示实验组中耳黏膜局部Foxp3mRNA表达OME组较对照组增高(P=0.0036<0.05);RORγt m RNA表达两者差异无统计学意义(P=0.1069>0.05)。结论OME的免疫相关发病机制之一可能是全身免疫反应与局部免疫反应的共同作用的结果,以中耳局部免疫反应为主;TGF-β参与OME全身免疫应答,CD4^+CD25^+Foxp3^+Treg细胞更倾向在中耳局部的免疫应答中发挥作用;FOXP3基因参与调控OME中耳局部免疫机制;Th17细胞在大鼠OME的免疫机制中的作用有待进一步研究。展开更多
The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible role...The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.展开更多
文摘目的:研究五虎汤对病毒诱发幼年哮喘大鼠脾细胞悬液T-bet、GATA-3、RORγt、Foxp3 m RNA表达的影响。方法:采用RSV诱导OVA致敏大鼠哮喘模型。造模成功后,随机分成五虎汤高、中、低[4.752 g/(kg·d)、2.376 g/(kg·d)、1.188 g/(kg·d)]剂量组、地塞米松组[地塞米松片,2.4 mg/(kg·d))]、模型组。干预14 d后,取脾组织,采用RT-PCR法检测脾细胞悬液中T-bet、GATA-3、RORγt、Foxp3 m RNA表达情况。结果:模型组脾组织T-bet、Foxp3m RNA表达低于空白组(P<0.05),而GATA-3、RORγt m RNA表达高于空白对照组(P<0.05)。五虎汤高、中剂量组脾组织GATA-3、RORγt m RNA表达均明显低于模型组(P<0.01或P<0.05),而T-bet、Foxp3 m RNA表达均明显高于模型组(P<0.01或P<0.05)。结论:五虎汤能调控病毒诱发幼年哮喘大鼠脾组织转录因子T-bet、GATA-3、RORγt、Foxp3 m RNA的表达,调控Thl、Th2、Thl7、Treg细胞,重建免疫平衡,抑制哮喘气道炎症形成。
基金This project was supported by a program of Science Project of Hubei Province (No.2003AA301C10).
文摘The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.
