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抗氧化劑 Cu/Zn SOD 和 Ascorbate 皆可減少 Pyrogallol 和 Duroquinone 抑制 SNAP 所引起的平滑肌鬆弛反應

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• 系統編號 RC9008-0161

• 計畫中文名稱 非膽鹼性系統對呼吸道/肺之調控---抗氧化劑對天竺鼠呼吸道之非膽鹼性神經系統的保護角色之調控(II)

• 計畫英文名稱 Antioxidant Protection of Inhibitory Non-Cholinergic Neurotransmitter in Guinea-Pig Trachea (II)

• 主管機關 行政院國家科學委員會 • 計畫編號 NSC89-2320-B038-017-M41

• 執行機構 台北醫學院醫事技術系

• 本期期間 8808 ~ 8907

• 報告頁數 5 頁 • 使用語言 中文

• 研究人員 林建煌 Lin, Chien Huang

• 中文關鍵字 抗氧化劑;非膽鹼性;一氧化氮;超氧歧化脢;神經傳遞物質

• 英文關鍵字 Antioxidant;Non-cholinergic;Nitric oxide (NO);Superoxidase dismutase;Neurotransmitter

• 中文摘要

Superoxide anion generator 如 Pyrogallol 和 Duroquinone 可抑制 SNAP 所引起的平滑肌鬆弛反應,但對於直接電刺激 NANC 神 經,所引起的平滑肌鬆弛反應則沒有抑制作用。抗氧化劑 Cu/Zn SOD 和 Ascorbate 皆可減少 Pyrogallol 和 Duroquinone 抑制 SNAP 所引起的平滑肌鬆弛反應。而 Pyrogallol 和 Duroquinone 的抑制作用可被大量的 Cu/Zn SOD 所對抗。在

Diethyldithiocarbamate (Cu/Zn SOD 抑制劑)存在下,Pyrogallol 和 Duroquinone 可抑制電刺激所引發氣管的鬆弛反應。另外,

Buthionine sulphoximine (1mM,An inhibitor of glutathione production),Etharynic acid (30μM,A sulphydryl alkylator),

Sulphobromophthalein (100μM,An inhibitor of glutathione S-transferase), Diamide (100μM,A glutathione oxidator)以劑量相關 的方式抑制電刺激 NANC 神經所引起的氣管平滑肌鬆弛反應。Carboxy-PTIO (NO scavengers)也以劑量相關的方式抑制 SNAP 所引起的平滑肌鬆弛反應,但對於電刺激 NANC 神經所引起的平滑肌鬆弛反應則沒有抑制作用。氣管平滑肌組織均質液具有 SOD 的活性,且 Mn SOD 及 Cu/Zn SOD 的活性分別佔 34.2%及 65.8%。由 Western bloting 的實驗方法可得知,天竺鼠氣管平 滑肌組織均質液內存在有 Mn SOD, Cu/Zn SOD, Catalase 三種抗氧化酵素。由 Immunocytochemistry 的實驗方法可得知,

神經附近可發現有 Mn SOD, Cu/Zn SOD 或 Catalase 三種抗氧化酵素的存在。因此神經附近的抗氧化酵素 Mn SOD, Cu/Zn SOD 或 Catalase 可藉由清除氧游離基,來減少 Peroxynitrite 對神經細胞之傷害性或延長 NO 之生物活性,因而在發炎的過程 中扮演保護抑制性 NANC 神經系統的功能。

• 英文摘要 There is evidence that nitric oxide (NO) as an prominent i-NANC neurotransmitter in the airway smooth muscle. The superoxide

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anion generator, pyrogallol and duroquinone reduced tracheal relaxations to SNAP, but had no effect on those to field stimulation.

The inhibitory effect of pyrogallol and duroquinone was partially reversed by Cu/Zn SOD and ascorbate. Following inhibition of Cu/Zn SOD activity with diethyldithiocarbamate, pyrogallol and duroquinone also reduced tracheal relaxations to field stimulation and this effect was again by reversed Cu/Zn SOD and ascorbate. Buthionine sulphoximine (1mM, an inhibitor of glutathione

production), etharynic acid (30μM, a sulphydryl alkylator), sulphobromophthalein (100μM, an inhibitor of glutathione S-transferase), diamide (100μM, a glutathione oxidator) reduced tracheal relaxations to field stimulation. Carboxy-PTIO (NO scavengers) also reduced tracheal relaxations to SNAP, but had no effect on those to field stimulation. Immunostainstaing demonstrated Mn SOD, Cu/Zn SOD, catalase immunoreactivity in the nerve terminal. Western blot analysis confirmed the presence of these enzyme in the homogenates of tracheal muscle, and enzyme assays demonstrated exist Mn SOD, and Cu/Zn SOD activities. Therefore antioxidant enzyme found in the nerves may play a role in regulating NO-mediated neuromuscular communication in the guineapig trachea.

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