雷公藤甲素对体外不同密度嗜酸细胞凋亡及GM-CSF受体αmRNA表达的影响

目的观察雷公藤甲素(Triptolide,TP)对哮喘豚鼠体外不同密度嗜酸细胞(Eos)凋亡及粒细胞-巨噬细胞克隆剌激因子受体α(GM-CSFRα)mRNA表达的影响,探讨TP促Eos凋亡的机制。方法卵蛋白激发哮喘豚鼠48h后,行支气管肺泡灌洗,分离低密度Eos(HEos)及正常密度Eos(NEos)。HEos及NEos分别与TP(10-7～10-4mol/L)共培养24h,以原位杂交检测Eos表达GM-CSFRαmRNA,3′末端脱氧核苷转移酶介导的脱氧三磷酸尿苷(d-UTP)缺口末端标记(TUNEL)法检测细胞凋亡。结果哮喘豚鼠不同密度Eos经TP干预24h后,细胞凋亡增加,与对照相比HEos在10-7mol/L时出现差异(P<0.05),NEos在10-6mol/L时出现差异(P<0.05),同时不同密度EosGM-CSFRαmRNA表达增加,与对照相比均在10-5mol/L时出现差异(P<0.05)。Eos凋亡与TP浓度呈剂量依赖性。HEos与NEos表达GM-CSFRα无差异(P>0.05)。不同密度Eos表达GM-CSFRα与Eos凋亡呈正相关(P<0.05)。结论TP可促进哮喘豚鼠不同密度Eos...     

雷公藤甲素抑制IL—5介导的人嗜酸性粒细胞存活延长作？…

目的：探讨雷公藤甲素（ＴＰ）对体外ＩＬ－５介导的人嗜酸性粒细胞（Ｅｏｓ）存活延长作用的时间。方法：在不同浓度ＩＬ－５和ＴＰ的条件下体外培养Ｅｏｓ，台盼蓝拒染法观察细胞活力的变化，结合原位末端标记和ＤＮＡ电泳检测Ｅｏｓ凋亡的变化。免疫组化检测ＴＰ刺激后Ｅｏｓ凋亡基因Ｆａｓ表达的变化。结果：ＩＬ－５显著延长Ｅｏｓ体外存活的时间，ＴＰ呈浓度和时间依赖性抑制ＩＬ－５介导的Ｅｏｓ存活延长作用，ＴＰ（１０＾－     

氨茶碱治疗对哮喘豚鼠肺内嗜酸性粒细胞凋亡？…

目的：探讨氨茶碱对哮喘肺内嗜酸性粒细胞凋亡的作用。方法：以卵白蛋白致敏的过敏性哮喘豚鼠 端标记原位细胞凋亡检测、光镜和电镜形态学方法。观察氨茶碱治疗对哮喘豚鼠肺内Ｅｏｓ凋亡的作用。结果：氨茶碱治疗导致哮喘豚鼠支气管肺泡灌洗液Ｅｏｓ数量显著降低，肺内Ｅｏｓ数量显著降低，肺内Ｅｏｓ凋亡显著增加，凋亡Ｅｏｓ能被巨噬细胞吞噬。结论：诱导的Ｅｏｓ凋亡是氨茶碱治疗哮喘的一个重要机制。     

雷公藤甲素抑制IL-5介导的人嗜酸性粒细胞存活延长作用及其机制研究

目的 :探讨雷公藤甲素 (TP)对体外IL 5介导的人嗜酸性粒细胞 (Eos)存活延长作用的影响。方法 :在不同浓度IL 5和TP的条件下体外培养Eos ,台盼蓝拒染法观察细胞活力的变化 ,结合原位末端标记和DNA电泳检测Eos凋亡的变化。免疫组化检测TP刺激后Eos凋亡基因Fas表达的变化。结果 :IL 5显著延长Eos体外存活的时间 ,TP呈浓度和时间依赖性抑制IL 5介导的Eos存活延长作用 ,TP(10 - 7mol L)显著抑制IL 5 (1 0U ml)介导的Eos的存活 ,原位凋亡检测和DNA电泳表明此作用是TP诱导Eos凋亡所致。TP使Eos的Fas表达显著增强。结论 :TP具有拮抗IL 5诱导Eos凋亡的作用 ,其作用机制可能与增强Eos的Fas表达有关。     

雷公藤多甙对哮喘患者痰液IL-5和嗜酸细胞凋亡的影响

目的　探讨雷公藤多甙 (TP)对哮喘患者的治疗作用。方法　选择 70例轻、中度哮喘缓解期患者 ,随机分成A组 (TP治疗组 )和B组 (对照组 ) ,治疗前后分别采用ELISA法测定患者痰液白细胞介素 5 (IL 5 )和可溶性白细胞介素 2受体 (sIL 2R) ,应用流式细胞仪检测痰液嗜酸细胞 (EOS)凋亡。结果　缓解期哮喘患者痰液中IL 5和sIL 2R明显升高。A组经TP治疗后 ,IL 5和sIL 2R明显降低 ,EOS凋亡率明显升高 ,B组治疗前后上述各项指标差异无显著性。结论　TP可能通过抑制哮喘患者T细胞和B细胞的活化 ,增加EOS的凋亡 ,从而减少机体免疫 炎症反应。     

雷公藤甲素对淋巴细胞表达IL─5mRNA的作用

从健康志愿献血员的外周血中分离出单个核细胞（ＰＢＭＣ），调细胞浓度为５×１０６／ｍｌ，加入终浓度为５μｇ／ｍｌ的ＣｏｎＡ和终浓度为０、１、１０、５０、１００ｎｇ／ｍｌ的雷公藤甲素，体外培养２４ｈ后收集培养的细胞，用异硫氰酸胍法提取总ＲＮＡ。将含小鼠ＩＬ－５ｃＤＮＡ的质粒酶切为约１ｋｂ的片段；用地高辛配基标记探针；用斑点印迹杂交法检测ＰＢＭＣ的ＩＬ－５ｍＲＮＡ表达。结果终浓度为１ｎｇ／ｍｌ和１０ｎｇ／ｍｌ的雷公藤甲素对ＰＢＭＣ表达ＩＬ－５ｍＲＮＡ无明显作用。而终浓度为５０ｎｇ／ｍｌ和１００ｎｇ／ｍｌ的雷公藤甲素能显著地抑制ＣｏｎＡ刺激的ＰＢＭＣ表达ＩＬ－５ｍＲＮＡ     

氨茶碱治疗对哮喘豚鼠肺内嗜酸性粒细胞凋亡的作用

目的：探讨氨茶碱对哮喘肺内嗜酸性粒细胞（Eos）凋亡的作用。方法：以卵白蛋白致敏的过敏性哮喘豚鼠为模型,采用3’－末端标记原位细胞凋亡检测、光镜和电镜形态学方法,观察氨茶碱治疗对哮喘豚鼠肺内Eos凋亡的作用。结果：氨茶碱治疗导致哮喘豚鼠支气管肺泡灌洗液Eos数量显著降低,肺内Eos凋亡显著增加,凋亡Eos能被巨噬细胞吞噬。结论：诱导肺内Eos凋亡是氨茶碱治疗哮喘的一个重要机制。     

哮喘豚鼠肺组织嗜酸粒细胞IL-5和GM-CSF mRNA的表达及药物治疗的影响

哮喘肺组织嗜酸粒细胞 (Ｅｏｓ)存活依赖于细胞因子ＩＬ 3、ＩＬ 5、ＧＭ ＣＳＦ的刺激。通常认为 ,Ｅｏｓ的存活因子主要来源于激活的Ｔ淋巴细胞。近年来研究表明 ,一些过敏性疾病和变态反应性疾病Ｅｏｓ亦能表达ＩＬ 3、ＩＬ 5、ＧＭ ＣＳＦ等多种细胞因子〔1〕。本研究采用过敏性哮喘豚鼠模型和组织细胞原位杂交 (ＩＳＳＨ)方法 ,对哮喘肺组织和支气管肺泡灌洗液 (ＢＡＬＦ)ＥｏｓＩＬ 5、ＧＭ ＣＳＦｍＲＮＡ的表达及其与凋亡的关系进行了研究。材料与方法动物模型与分组 :正常对照组 ;哮喘对照组用卵白蛋白致敏和激发哮喘 ;药物治疗组…     

白介素5与嗜酸性粒细胞凋亡

Ｉ Ｌ５ 抑制或延迟嗜酸性粒细胞凋亡,导致嗜酸性粒细胞在组织中聚积。这与ｂｃｌ２ 表达增多、酪氨酸激酶活化、 Ｒ Ｎ Ａ 及蛋白质的合成受抑制有关。嗜酸性粒细胞凋亡与巨噬细胞功能有一定关系。     

Increased chemokinetic and chemotactic responses of eosinophils in asthmatic patients

The aim of the present study was to investigate the migratory responses of eosinophil and neutrophil granulocytes from asthmatic patients compared with granulocytes from healthy individuals. Twenty-three patients with unstable and severe asthma and blood eosinophilia (greater than 400 x 10(6) cells/l) were selected for the study. Eosinophil and neutrophil chemotactic and chemokinetic responses were tested twice, at the beginning and end of a 5 week treatment period. Lung function was followed by daily measurements of PEF. The eosinophils of the asthmatics demonstrated increased chemokinetic responses to albumin, autologous serum, and normal human serum (NHS), and an increased chemotactic response to NHS at the beginning of the treatment period compared with eosinophils from the references. At the end of the period the eosinophil chemokinetic responses to albumin, autologous serum and NHS were still increased and so was the chemotactic response to zymosan-activated serum (ZAS). The neutrophil migratory responses were not increased compared with those of the references, except for the chemokinetic response to autologous serum, which was increased both at the beginning and end of the treatment period. Patients in whom the eosinophil migratory responses, to most of the agents used, decreased over the treatment period, demonstrated a significantly greater improvement of their lung function at the end of the period compared with patients in whom the eosinophil migratory responses increased. However, no direct relationship between eosinophil migratory responses and lung function of the patients was found. In conclusion, the present investigation demonstrated increased migratory responses of eosinophils from asthmatic patients. This enhanced responsiveness is proposed to be due to priming of the eosinophils in vivo, and might be one mechanism behind the selective recruitment of eosinophils to the lungs of asthmatics.     

地塞米松对哮喘豚鼠嗜酸细胞凋亡及bcl—2 mRNA表达的影响

目的 观察地塞米松（DM）对哮喘豚鼠体外不同密度的嗜酸性粒细胞（Eos)凋亡及bcl-2表达的影响，并探讨激素促进哮喘Eos凋亡的机制。方法 以卵蛋白激发哮喘豚鼠模型48h后，进行支气管肺泡灌洗，分离灌洗液中不同密度的Eos。加入DM（10^10-10^5mol/L），以原位杂交法检测Eos的bcl-2表达，TUNEL法检测Eos的凋亡。结果 DM干预24h，可见Eos凋亡增加，bcl-2 mRNA表达降低，且呈剂量依赖性。bcl-2 mRNA的表达与Eos凋亡呈负相关。结论 DM可抑制bcl-2的表达，促进Eos凋亡。bcl-2表达的减少可能是DM调节Eos凋亡的机制之一。     

哮喘豚鼠IL-5、IL-3、GM-CSF mRNA表达及雷公藤内酯醇的影响

目的研究ＩＬ－５、ＩＬ－３、ＧＭ－ＣＳＦ在哮喘发病中的作用及雷公藤的干预。方法将实验豚鼠随机分为：①哮喘组（ｎ＝８）：用卵蛋白雾化吸入诱导哮喘模型；②处理组（ｎ＝８）：用雷公藤内酯醇腹腔注射处理哮喘模型；③正常对照组（ｎ＝８）。制备ＩＬ－５、ＩＬ－３和ＧＭ－ＣＳＦｃＤＮＡ探针，用斑点印迹杂交法检测以上三组豚鼠支气管肺组织ＩＬ－５、ＩＬ－３和ＧＭ－ＣＳＦｍＲＮＡ的表达。结果哮喘豚鼠支气管肺组织ＩＬ－５、ＩＬ－３和ＧＭ－ＣＳＦｍＲＮＡ表达显著高于对照组（Ｐ＜０．０５～０．００１）；雷公藤处理组ＩＬ－５、ＩＬ－３和ＧＭ－ＣＳＦｍＲＮＡ表达低于哮喘组（Ｐ＜０．０５～０．００１），与对照组无显著差异（Ｐ＞０．０５）。结论哮喘豚鼠肺组织中有明显的ＩＬ－５、ＩＬ－３和ＧＭ－ＣＳＦｍＲＮＡ表达增加。雷公藤内酯醇能抑制体内ＩＬ－５、ＩＬ－３和ＧＭ－ＣＳＦｍＲＮＡ的表达，可能在哮喘抗炎中具有潜在的临床价值。     

地塞米松对哮喘豚鼠肺组织嗜酸细胞bcl-2表达的影响及意义

目的 研究哮喘豚鼠肺内不同密度嗜酸细胞（Eos）凋亡与bcl-2 mRNA表达的关系及地塞米松（DM）对它们的影响。方法 应用DM干预哮喘豚鼠,分离哮喘豚鼠支气管肺泡灌洗液中不同密度Eos,以TUNEL法检测细胞凋亡,以原位杂交检测bcl-2 mRNA表达。结果 哮喘组Eos凋亡率较正常组明显降低（P〈0．01）,应用DM后均显著增加（P〈0．01）。正常豚鼠Eos可检测到bcl-2 mRNA表达,不同密度Eos表达无显著差异（P〉0．05）。哮喘组bcl-2mRNA明显增加,应用DM后其表达明显减少。结论凋亡调节的缺失是导致组织及血中Eos增多的重要原因,bcl-2参预了哮喘Eos凋亡的调节。DM可促进哮喘肺组织中的Eos细胞凋亡,bcl-2可能是DM调节Eos细胞凋亡的重要途径之一。     

Down-regulated IL-5 receptor expression on peripheral blood eosinophils from budesonide-treated children with asthma

BACKGROUND: The expression and function of cytokine receptors on peripheral blood eosinophils (PBE) from healthy and asthmatic children are poorly characterized. METHODS: The PBE count and expression of IL-5 receptor (R) and GM-CSFR positive PBE was analyzed in nonsteroid-treated asthmatic children (n = 13), budesonide-treated asthmatic children (n = 24) and healthy children (n = 16) by flow cytometry. Alterations in intracellular EG2-epitope expression were used to measure the in vitro responsiveness of PBE to recombinant IL-5 and GM-CSF. RESULTS: The PBE count was increased (P < 0.05) in both asthmatic groups, independent of treatment, as compared to healthy children. The IL-5R expression on PBE, as well as the in vitro responsiveness of PBE to recombinant IL-5, was reduced (P < 0.05), in budesonide-treated asthmatic children compared to nonsteroid-treated asthmatic children and healthy children. The proportion of GM-CSFR positive PBE and in vitro responsiveness of PBE to recombinant GM-CSF were not different between the groups. In vitro treatment with budesonide did not down-regulate the proportion of IL-5R positive PBE. CONCLUSIONS: Budesonide-treatment of asthmatic children induces a selectively reduced IL-5R expression on PBE, concomitant with a reduced in vitro responsiveness of PBE to IL-5. We suggest that this budesonide-related down-regulation of the IL-5R might be a mechanism by which steroid treatment inhibits the action of IL-5 on eosinophil accumulation and activation in vivo.     

Dendritic cell number is related to IL-4 expression in the airways of atopic asthmatic subjects

BACKGROUND: Airway dendritic cells are essential for stimulating naive T cells in response to inhaled antigen and for the development of allergic sensitization. IL-4 in vitro can distinguish dendritic cell lines from peripheral blood mononuclear cells. Our study had the following aims: 1) to compare the distribution of CD1a+ dendritic cells and IL-4+ cells, in the bronchial mucosa of asthmatics and controls 2) to determine the relationship between the numbers of CD1a+ dendritic cells and IL-4+ cells in the bronchial mucosa of asthmatics 3) to determine whether CD1a+ cells express the IL-4 receptor. METHODS: Twenty atopic asthmatic and eight normal subjects were studied. In each subject, bronchoscopy with bronchial biopsies was performed. CD1a, IL-4, and IL-4 receptor expressions were evaluated by immunohistochemistry. RESULTS: The number of CD1a+ and IL-4+ cells was significantly higher in asthmatics than controls. The number of CD1a+ cells was positively correlated to the number of IL-4 + cells. Bronchial biopsy serial section studies showed that CD1a+ cells express the receptor for IL-4. CONCLUSIONS: These results suggest that an increased amount of IL-4 may play a physiopathologic role in maintaining the dendritic cell pool in vivo. Therefore, because of possible IL-4 activity on antigen-presenting cells in T-cell immune responses to allergens, an important new role of IL-4 in asthma inflammation can be envisaged.     

地塞米松对哮喘豚鼠肺及内脏感觉传入系统神经生长因子表达的抑制作用

目的：探讨地塞米松对哮喘豚鼠肺及内脏感觉传人系统(C7-T5脊神经节及对应的脊髓后角)神经生长因子(NGF)表达的影响。方法：利用免疫组织化学和Westem印迹方法,观察生理盐水组、单纯致敏组、哮喘组和地塞米松组豚鼠肺及内脏感觉传人系统(C7-T5脊神经节及对应的脊髓后角)NGF表达的变化。结果：哮喘组豚鼠NGF在肺及内脏感觉传人系统(C7-T5脊神经节及对应的脊髓后角)表达明显高于对照组,而地塞米松组则明显低于哮喘组。结论：NGF可能参与哮喘的发病过程,地塞米松抑制哮喘豚鼠NGF的表达可能是其治疗哮喘的机制之一。     

吸入地塞米松对哮喘豚鼠内皮素-1的影响

建立豚鼠哮喘模型,检测肺活组织、支气管肺泡灌洗液（ＢＡＬＦ）、血浆内皮素－１（ＥＴ－１）水平有地塞米松干预后的变化以探讨ＥＴ－１在支气管的作用。结果显示哮喘组肺活组织、ＢＡＬＦ、血浆ＥＴ－１显著升高：吸入地塞米松后ＥＴ－１均显著下降。提示：ＥＴ－１可能参与支气管哮喘的发生和 ,吸入糖皮质激素可以抑制体内ＥＴ－１的生成和释放,这可能是糖皮质激素治疗支气管哮喘的重要机制之一。     

雷公藤对致敏小鼠T细胞IL-5 mRNA表达及转录因子GATA-3活性的抑制作用

探讨雷公藤内酯醇 (TP )对致敏小鼠T淋巴细胞IL 5mRNA表达的影响及其机制。采用卵蛋白 (OVA )致敏的方法建立模型 ;运用原位杂交染色法 (ISH )观察TP对T淋巴细胞IL 5mRNA表达的影响 ;通过凝胶电泳迁移率实验 (EMSA )对CD4+T淋巴细胞核转录因子GATA 3的DNA结合活性进行检测 ,同时就TP的作用与地塞米松 (DM )相比较。结果表明致敏小鼠T淋巴细胞IL 5mRNA表达显著高于正常对照组 (P <0 0 1) ,经TP、DM处理后 ,其IL 5mRNA表达显著低于致敏组(P <0 0 1)。致敏小鼠CD4+ T淋巴细胞体外经伴刀豆蛋白A (ConA )刺激后 ,GATA 3的DNA结合活性与正常对照组比较显著增强 ,并呈时间依赖关系 ,经TP、DM处理后 ,GATA 3的DNA结合活性显著减弱。TP抑制IL 5基因转录的分子机制可能与其抑制GATA 3的DNA结合活性有关。     

Effect of inhaled interleukin-5 on eosinophil progenitors in the bronchi and bone marrow of asthmatic and non-asthmatic volunteers

BACKGROUND: Asthma is characterized by increases in mature eosinophils and their progenitors within the bronchus and bone marrow. IL-5 plays a key role in eosinophil development in the bone marrow and at the site of allergic inflammation. We therefore studied the effects of nebulized IL-5 on eosinophils, their progenitors and in situ haemopoiesis within the airway and bone marrow. METHODS: Nine atopic asthmatics and 10 non-atopic non-asthmatic control volunteers inhaled 10 microg of IL-5 or placebo via a nebulizer in a double-blind, randomized, cross-over study. Bronchoscopy, bone marrow aspiration and peripheral blood sampling were performed 24 h after nebulization. Four weeks later, volunteers inhaled the alternative solution and underwent a repeat bronchoscopy and bone marrow aspiration. RESULTS: Inhalation of IL-5 significantly decreased CD34(+)/IL-5Ralpha mRNA(+) cells within the bronchial mucosa and the percentage of CD34(+) cells that were CCR3(+) within the bone marrow of atopic asthmatic, but not control, volunteers. Inhalation of IL-5 also induced a significant increase in bronchial mucosal eosinophils in the non-atopic non-asthmatic control volunteers, but not in the asthmatics. IL-5 had no effect on spirometry or airways hyper-reactivity in either group. CONCLUSIONS: Inhaled IL-5 modulated eosinophil progenitor numbers in both the airways and bone marrow of asthmatics and induced local eosinophilia in non-asthmatics.