Effect of eosinphil granule proteins on beta-adrenergic receptor

嗜酸性颗粒蛋白对β-肾上腺素能受体的影响

• 批准号: 02670346
• 负责人: MOTOJIMA Shinji
• 金额: $ 0.64万
• 依托单位: Dokkyo University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670346/
• 关键词: beta-adrenergic receptor; eosinphil peroxidase; major basic protein; eosinphil cationic protein; lung; S49lymphoma cell; cyclic AMP; 肺; cyclic AMP; 好酸球; eosinophil peroxidase

Effect of eosinophil peroxidase (EPO) + H_2O_2 on beta-adrenergic receptor number was studied. Human EPO was obtained from eosinophils of hypereosinophilic syndrome patients by sequential chromatograph on Sephadex G-50 and CM-Sepharose column. Guinea pig lung membrane fraction was first preincubated with EPO, then incubated with H_2O_2 for 2 hours. The number of beta-adrenergic receptor was measured by binding assay. The number of beta adrenergic receptor was significantly decreased by 3-10 U/ml EPO and 10^<-4>M H_2O_2. Beta-receptor number was also decreased by 10 U/ml EPO and 10^<-6>M H_2O_2.Next effect of EPC on beta-adrenergic receptor-adenylate cyclase system was studied. S 49 cells were preincubated with EPO for 15 minutes, and then incubated with 10^<-4>M H_2O_2 for 45 minutes. The cells were centrifuged, resuspended in culture medium including xanthine, then stimulated by 10^<-4>M isoproterenol to generate intracellular cyclic AMP (CAMP). Intracellular cyclic AMP production wap … More significantly inhibited by the pretreatment with 0.1-1 U/ml EPO and 10^<-3>M H_2O_2, and the degree of inhibition was EPO concentration dependent. CAMP production was inhibited by 91 % by pretreatment with 1 U/ml FPO and 10^<-3>M H_2O_2. CAMP production was also inhibited when 10^<-4>M or 10^<-5>M H_2O_2 was applied with EPO. EPO or H_2O_2 alone did not inhidited CAMP production. The number of beta-adrener gic receptor on S49 cells did not change by EPO + H_2O_2 system, however, the affinity of beta-adrenergic receptor to isoproterenol was decreased. CAMP production stimulated by 10^<-5>M forskolin was also inhibited by EPO + H_2O_2, however, the degree of inhibition was greater in isoproterenol stimulation than in forskolin stimulation.Major basic protein and eosinophil cationic protein did not have significant suppressive effects either on beta-adrenergic receptor number or on CAMP production.These results suggest that EPO is one of the factors that induce betaadrenergic hyporesponsiveness in patients with bronchial asthma. Less

本文研究了嗜酸性粒细胞过氧化物酶（EPO）+ H_2O_2对β-肾上腺素能受体数目的影响。采用SephadexG-50和CM-Sepharose柱连续层析法从高嗜酸性粒细胞综合征患者的嗜酸性粒细胞中分离出人EPO。用EPO预孵育豚鼠肺细胞膜，再用H_2O_2孵育2 h。通过结合试验测定β-肾上腺素能受体的数量。3-10 U/ml EPO和10 μ M H_2O_2可明显减少β-肾上腺素能受体的数量<-4>。10 U/ml EPO和10 μ M H_2O_2也可使β受体数目减少<-6>。用EPO预孵育S49细胞15分钟，再用10 μ <-4>M H_2O_2孵育45分钟。将细胞离心，重悬于包含黄嘌呤的培养基中，然后用10 μ <-4>M异丙肾上腺素刺激以产生细胞内环AMP（CAMP）。细胞内cAMP生成 ...更多信息 0.1- 1U/ml EPO和10 μ M H_2O_2预处理可显著抑制<-3>细胞增殖，且抑制程度呈EPO浓度依赖性。用1U/ml FPO和10 μ M H_2O_2预处理对cAMP的产生抑制91%<-3>。10 μ <-4>M或10 μ <-5>M H_2O_2与EPO合用也能抑制cAMP的产生。EPO或H_2O_2单独使用不能抑制cAMP的产生。EPO + H_2O_2系统对S49细胞β-肾上腺素能受体的数量无明显影响，但能降低β-肾上腺素能受体对异丙肾上腺素的亲和力。EPO + H_2O_2对10 μ M Forskolin刺激的cAMP产生<-5>也有抑制作用，但对异丙肾上腺素刺激的抑制程度大于Forskolin刺激，而主要碱性蛋白和嗜酸性粒细胞阳离子蛋白对β-肾上腺素能受体数目和cAMP产生均无明显抑制作用，提示EPO是引起哮喘患者β-肾上腺素能低反应性的因素之一。少

项目成果

本島 新司: "細胞内情報伝達系と疾患,気管支喘息" クリニカル・ニュ-ロサイエンス. 9. 888-890 (1991)

Shinji Motojima：“细胞内信号转导系统和疾病，支气管哮喘”《临床神经科学》9. 888-890 (1991)。

本島 新司,牧野 荘平: "レセプタ-,気管支喘息とβアドレナリンレセプタ-" 朝倉書店, (1992)

本岛真司、牧野翔平：“受体、支气管哮喘和 β-肾上腺素能受体”朝仓书店，(1992)

本島 新司,牧野 荘平: "好酸球顆粒蛋白のモルモット肺β受容体に対する影響" アレルギ-.

Shinji Motojima、Shohei Makino：“嗜酸性粒细胞颗粒蛋白对豚鼠肺 β 受体的影响”过敏。

本島 新司,牧野 荘平: "Eosinophil peroidaseのβ受容体・adenylate cyclase" 日本胸部疾患学会雑誌.

Shinji Motojima、Shohei Makino：“嗜酸性粒细胞过氧化物酶和腺苷酸环化酶的β受体”日本胸部疾病学会杂志。

Shinji Motojima, Sohei Makino: "Effect of eosinhphil granule proteins on beta-adrenergic receptors in guinea pig lungs." Jpn. J. Allergol.

Shinji Motojima、Sohei Makino：“嗜酸性颗粒蛋白对豚鼠肺中 β-肾上腺素能受体的影响。”