Molecular biological analysis of bradykinin B1 receptor in chronic inflammatory pulmonary diseases.

慢性炎症性肺部疾病缓激肽 B1 受体的分子生物学分析。

• 批准号: 11670565
• 负责人: TSUKAGOSHI Hideo
• 金额: $ 1.09万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670565/
• 关键词: chronic pulmonary diseases; idiopathic pulmonary fibrosis; bradykinin receptor; kinin-kallikrein system; lung fibroblast; extracellular matrix; cyclooxygenase; polymorphism; ブラジキニンB_1受容体; 炎症性肺疾患; 分子生物学的解析

To investigate the role of kinin-kallikein system in chronic inflammatory pulmonary diseases, we studied the molecular biological function of bradykinin B1 receptor in vitro and ex vivo. Bradykinin receptor has two subtypes B1 and B2, which are inducible and constitutive receptors for des-Arg^9-bradykinin and des-Arg^10-kallidin and for bradykinin and kallidin, B1 and B2 receptor agonists, respectively. First we characterized murine B1 receptor in a murine alveolar macrophage cell line MH-S. Pro-inflammatory cytokine interleukin-1__.β(IL -1__.β)induced murine B1 receptor gene expression assessed by quantitative real time RT-PCR, and B1 agonist stimulated the production of tumor necrosis factor alpha (TNF-α) by IL-1__.β-pretreated MH-S cells. Then we characterized human Bl receptor in human lung fibroblast cell line IMR-90. IL-1__.βinduced human Bl receptor gene expression assessed by quantitative real time RT-PCR, and Bl agonist stimulated the production of extracellular matrix fibronectin by IMR-90 cells. With Bl receptor agonist stimulation IL-1__.β-pretreated IMR-90 cells produced less fibronectin than that of IL-1__.β-untreated IMR-90 cells. This interesting phenomenon was reversed by cyclooxygenase (COX) inhibitors indomethacin and NS-398, the dual COX1/COX-2 inhibitor and selective COX-2 inhibitor, respectively. Furthermore, there was a decrease in the gene expression of B1 receptor in the lung tissue of idiopathic pulmonary fibrosis patients compared to the control. These results suggest that kinin-kallikrein system play some role in the chronic inflammatory pulmonary diseases via B1 receptor activation to protect lung fibrosis in combination with inducible COX-2. Because there are some polymorphisms in B1 receptor gene in other diseases, further investigation would be needed in chronic inflammatory pulmonary diseases as well especially in idiopathic pulmonary fibrosis.

为探讨激肽-激肽系统在慢性炎症性肺疾病中的作用，本研究对缓激肽B1受体的分子生物学功能进行了体外和离体研究。缓激肽受体有两种亚型B1和B2，分别是des-Arg^9-缓激肽和des-Arg^10-胰激肽的诱导型和组成型受体，以及缓激肽和胰激肽的B1和B2受体激动剂。首先，我们在小鼠肺泡巨噬细胞系MH-S中表征了小鼠B1受体。用真实的时间定量RT-PCR检测促炎细胞因子白细胞介素1 β（IL-1_. β）诱导小鼠B1受体基因表达，B1受体激动剂刺激IL-1_.β预处理的MH-S细胞产生肿瘤坏死因子α（TNF-α）。然后，我们表征了人肺成纤维细胞系IMR-90中的人B1受体。通过定量真实的RT-PCR评估IL-1 β诱导人B1受体基因表达，并且B1激动剂刺激IMR-90细胞产生细胞外基质纤连蛋白。在B1受体激动剂刺激下，IL-1_β预处理的IMR-90细胞产生的纤维连接蛋白比IL-1_β未处理的IMR-90细胞少。环氧化酶（考克斯）抑制剂吲哚美辛（indomethacin）和NS-398（COX 1/考克斯-2双重抑制剂和选择性考克斯-2抑制剂）分别逆转了这一有趣的现象。此外，与对照组相比，特发性肺纤维化患者肺组织中B1受体的基因表达降低。提示激肽释放酶系统可能通过激活B1受体与诱导型考克斯-2联合作用，在慢性炎症性肺疾病中发挥一定的保护肺纤维化作用。由于B1受体基因在其他疾病中也存在一定的多态性，因此在慢性炎症性肺疾病尤其是特发性肺纤维化中的研究还需进一步深入。

项目成果

Tsukagoshi H, Shimizu Y, Kawata T, Hisada T, Shimizu Y, Iwamae S, Ishizuka T, Iizuka K, Dobashi K, Mori M.: "Atrialnatriuretic peptide inhibits tumor necrosis factor-alpha production by interferon-gamma-activated macrophages via suppression of p38 mitogen

Tsukagoshi H、Shimizu Y、Kawata T、Hisada T、Shimizu Y、Iwamae S、Ishizuka T、Iizuka K、Dobashi K、Mori M.：“心钠素通过抑制干扰素-γ 激活的巨噬细胞抑制肿瘤坏死因子-α 的产生

塚越秀男ほか: "気道リモデリング形成に於ける酸化的ストレスの関与について"呼吸. 21(2). S (2002)

Hideo Tsukagoshi 等人：“氧化应激参与气道重塑”呼吸 21(2)。

Kasahara T,Tsukagoshi H, et al.: "PTH-rP-producing lung can cer with pso as abscess-like met astasisand humoral hypercal cemia of malignancy."Int J Clin Oncol. 5. 410-413 (2000)

Kasahara T、Tsukagoshi H 等人：“产生 PTH-rP 的肺癌，伴有脓肿样转移和恶性肿瘤的体液高钙血症。”Int J Clin Oncol。

Tsukagoshi H, Kawata T, Shimizu Y, Ishizuka T, Dobashi K, Mori M.: "4-hydroxy-2-nonenal enhances fibronectin production by IMR-90 human lung fibroblasts partly via activation of epidermal growth factor receptor-linked extracellular signal-regulated kinase

Tsukagoshi H、Kawata T、Shimizu Y、Ishizuka T、Dobashi K、Mori M.：“4-羟基-2-壬烯醛部分通过激活表皮生长因子受体相关的细胞外信号来增强 IMR-90 人肺成纤维细胞的纤连蛋白产生-

Tsukagoshi H, Shimizu Y, Iwamae S, Hisada T, Ishizuka T, Iizuka K, Dobashi K, Mori M.: "Evidence of Oxidative Stress in Asthma and COPD : Potential Inhibitory Effect of Theophylline"Resp Med. 94. 584-588 (2000)

Tsukagoshi H、Shimizu Y、Iwamae S、Hisada T、Ishizuka T、Iizuka K、Dobashi K、Mori M.：“哮喘和慢性阻塞性肺病氧化应激的证据：茶碱的潜在抑制作用”Resp Med。