Studies on pain measurements by the confocal laser scanning microscopy

共聚焦激光扫描显微镜疼痛测量的研究

• 批准号: 13671620
• 负责人: KUROSAKI Akiko
• 金额: $ 2.24万
• 依托单位: Osaka Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671620/
• 关键词: Allodynia; Knockout mice; Prostaglandin; Nitric oxide; Nociceptin; orphanin FQ; グルタミン酸; HIV gp120; NMDA受容体; カプサイシン; PGE受容体(EP); EPS受容体拮抗薬; 痛み; 共焦点顕微鏡; 微小循環; 共焦点レーザー生体顕微鏡; レチノイン酸; レチノール; comedolysis; 皮膚微小循環; 血管拡張薬

Functional evidence for interaction between prostaglandin EP3 and κ-opioid pathways in tactile pain induced by human immunodeficiency virus type-1 (HIV-1) glycoprotein gp120HIV-1 glycoprotein gp120 administrated intrathecally induces tactile pain (allodynia) in animals. In the present study, we investigated the mechanism of gp120-induced allodynia. Gp120 evoked allodynia in a dose-dependent manner with the maximum effect at 1 pg/mouse. Gp120 stimulated a rapid increase in intracellular free Ca^<2+> concentration ([Ca^<2+>]i) in the dorsal horn cells of the spinal cord. The gp120-induced allodynia was attenuated by indomethacin that inhibits prostaglandin synthesis, and did not develop in mice lacking PGE receptor EP3 subtype (EP3^<-/->). Pretreatment of spinal slices with indomethacin dose-dependently decreased the percentage of the cells which increased [Ca^<2+>]i in response to gp120, and the decrease was reversed by addition of the selective EP3 agonist. The κ-opioid agonist U-50,48 … More 8 significantly enhanced the gp120-stimulated increase in [Ca^<2+>]i in spinal slices prepared from EP3^<-/-> mice and simultaneous addition of U-50,488 with gp120 reproduced the gp120-induced allodynia in EP3^<-/-> mice. These results suggest that gp120 induces allodynia by increasing [Ca^<2+>]i followed by activation of EP3 and κ-opioid receptors in the spinal cord.Functional characterization of prostaglandin F_2α receptor in the spinal cord for tactile pain (allodynia)PGF_2α binds to its receptor (FP) to increase [Ca^<2+>]i by coupling of FP with G_qprotein. Spinal intrathecal administration of PGF_2α to mouse induces allodynia. FP in the spinal cord, however, was not well characterized. Here, we showed constitutive expression of FP mRNA in mouse spinal cord, and functionally characterized spinal FP-expressing cells which were involved in PGF_2α-induced allodynia. We identified an antisense oligodeoxyribonucleotide targeting FP mRNA, causing both disappearance of PGF_2α-induced allodynia and decrease of FP mRNA. With saline-administered mice, PGF_2α rapidly increased [Ca^<2+>]i of the cells in the deeper layer of the dorsal horn. In contrast, when the FP antisense oligodeoxyribonucleotide was repeatedly administered, the population of PGF_2α-responsive cells in the slices reduced, and PGF_2α-induced [Ca^<2+>]i increase of these cells diminished. These data strongly suggested that, in the dorsal horn of the spinal cord, there are the FP-expressing cells which are involved in PGF_2α induced allodynia. Less

前列腺素EP 3和κ-阿片途径在人类免疫缺陷病毒1型（HIV-1）糖蛋白gp120诱导的触觉疼痛中相互作用的功能证据鞘内注射HIV-1糖蛋白gp120诱导动物触觉疼痛（异常性疼痛）。在本研究中，我们探讨了gp120诱导异常性疼痛的机制。GP 120以剂量依赖性方式诱发异常性疼痛，最大效应为1 pg/小鼠。Gp 120刺激脊髓背角细胞内游离Ca ^2+浓度（[Ca ^2 +] i）迅速增加。gp120诱导的异常性疼痛被抑制前列腺素合成的吲哚美辛减弱，并且在缺乏PGE受体EP3亚型（EP3 ^<-/->）的小鼠中不发生。用吲哚美辛预处理脊髓切片，剂量依赖性地降低了响应gp120而增加[Ca ^2 +] i的细胞的百分比，并且这种降低被加入选择性EP 3激动剂逆转。κ-阿片激动剂U-50，48 ...更多信息 8显著增强了由EP 3 ^<-/->小鼠制备的脊髓切片中gp120刺激的[Ca ^<2 +>] i的增加，同时加入U-50，488和gp120在EP 3 ^<-/->小鼠中再现了gp120诱导的异常性疼痛。结果提示，gp120通过增加[Ca ^<2 +>] i，激活脊髓内EP 3和κ-阿片受体而引起触诱发痛，脊髓内前列腺素F_2 α受体在触觉痛（触诱发痛）中的功能特征PGF_2 α通过与G_q蛋白偶联而与其受体（FP）结合，增加[Ca ^<2 +>] i。脊髓鞘内注射PGF_2 α诱导小鼠痛觉超敏。然而，脊髓中的FP没有得到很好的表征。本研究显示FP mRNA在小鼠脊髓中的组成性表达，并对参与PGF_2 α诱导的异常性疼痛的脊髓FP表达细胞进行了功能鉴定。我们发现了一种靶向FP mRNA的反义寡核苷酸，它能使PGF_2 α诱导的痛觉异常消失，并使FP mRNA减少。在生理盐水组，PGF_2 α能迅速增加背角深层细胞的[Ca ^<2 +>] i。相反，反复给予FP反义寡核苷酸后，脑片中PGF_2 α反应性细胞的数量减少，PGF_2 α诱导的[Ca ^<2 +] i升高也减弱。这些结果表明，在脊髓背角内存在FP表达细胞，它们参与了PGF_2 α诱导的痛觉异常。少

项目成果

Mabuchi, T., et al.: "Attenuation of neuropathic pain by the nociceptin/orphanin FQ antagonist JTC-801 is mediated by inhibition of nitric oxide production"Eur.J.Neurosci.. 17. 1-9 (2003)

Mabuchi, T., et al.：“伤害感受肽/孤啡肽 FQ 拮抗剂 JTC-801 对神经性疼痛的减弱是通过抑制一氧化氮产生来介导的”Eur.J.Neurosci.. 17. 1-9 (2003)

伊藤誠二, 他: "脳機能の解明 -生命科学の主潮流- 遺伝子欠損マウスを用いた痛みの行動解析の現状と問題点(編:赤池紀扶, 東 英穂, 阿部康二, 久保千春)"ガイア出版会. 6 (2002)

Seiji Ito 等人：“大脑功能的阐明 - 生命科学的主要趋势 - 使用基因缺陷小鼠进行疼痛行为分析的现状和问题（编辑：Norifu Akaike、Hideho Higashi、Koji Abe、Chiharu Kubo）” Gaia出版。6（2002）

伊藤 誠二 他: "病態としての"痛み"研究:脊髄のPGsと痛み"脳. 21(6). 48-54 (2003)

Seiji Ito 等人：“疼痛作为一种病理状况的研究：PG 和脊髓疼痛”Brain 21(6) (2003)。

Minami, T.et al.: "Functional evidence for interaction between prostaglandin EP_3 and κ-opioid pathways in tactile pain induced by human immunodeficiency virus type-1 (HIV-1) glycoprotein gp120."Neuropharmacology. 45. 96-105 (2003)

Minami, T. 等人：“人类免疫缺陷病毒 1 型 (HIV-1) 糖蛋白 gp120 引起的触觉疼痛中前列腺素 EP_3 和 κ-阿片类药物通路之间相互作用的功能证据。” 45. 96-105 (2003) ）

南 敏明 他: "痛みの基礎と臨床"痛覚伝達に対する脊髄でのブロスタグランジンの役割. 11 (2003)

Toshiaki Minami 等人：“基本和临床疼痛” 脊髓中的溴前列腺素对疼痛传递的作用 11 (2003)。