甘藷胰蛋白抑制因子於巨噬細胞模式中對一氧化氮和活性氮化物之影響

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题名:	甘藷胰蛋白抑制因子於巨噬細胞模式中對一氧化氮和活性氮化物之影響
作者:	黃冠中(Guan-Jhong Huang)
贡献者:	藥學院中國藥學研究所
关键词:	甘藷;胰蛋白脢抑制因子;抗氮化;Peroxynitrite;Nitric oxide;Superoxide;Trypsin inhibitor;Sweet potato
日期:	2007-06-31
上传时间:	2009-09-01 16:03:00 (UTC+8)
摘要:	Peroxynitrite (ONOO-) 的產生是由superoxide 和 nitric oxide 反應而形成的，是最有影響細胞毒性的一種自由基，其可以氧化細胞組成包括蛋白質、酯類和DNA。 Peroxynitrite 可誘導細胞和組織損傷，結果導致數種疾病如: Alzheimer's disease, atherosclerosis 和 stroke. 由於?內缺乏清除 ONOO-的酵素，發現可清除ONOO- 的酵素是很重要的。本研究是利用甘藷分離出的胰蛋白脢抑制因子(TI)研究是否具有清除˙ON和 ONOO-的能力。由結果可知 TI 具有隨劑量增加清除nitrite和 superoxide自由基的能力。TI 清除superoxide 自由基的 IC50 值為 143.2 ±4.29 μg╱mL。我們使用 SOD 活性染色法證實TI 具有 SOD 活性.TI 也具有隨劑量增加而減低Peroxynitrite 氧化dihydrorhodamine 123 (DHR) 的能力。TI減低 Peroxynitrite 氧化 DHR 的IC50值為809.1 ± 32.36 μg╱mL。而利用分光光度計分析 TI 具有抑制ONOO—間接藉由電子傳遞機制來修飾 tyrosin 硝化作用。更進一步的研究發現，TI 具有抑制bovine serumalbumin (BSA)的硝化作用。TI 在巨嗜細胞中具有抑制 LPS-誘導nitrite 的產生且隨著濃度的增加而增加，其 IC50 值為 932.8± 29.85 μg╱mL。以目前的研究建議 TI 具有清除 reactive nitrogen species scavenging的能力. TI 也許具有清除 NO 和 ONOO-潛力可用以預防相關疾病. Peroxynitrite (ONOO-), formed from a reaction of superoxide and nitric oxide, is one of the most potent cytotoxic species that are known to oxidize cellular constituents including essential proteins, lipids, and DNA. ONOO- induces cellular and tissue injury, resulting in several human diseases such as Alzheimer's disease, atherosclerosis and stroke. Due to the lack of endogenous enzymes responsible for ONOO- scavenging activity, finding a specific ONOO- scavenger is of considerable importance. In this study, the ability of trypsin inhibitor (TI), isolated from sweet potato storage roots, to scavenge ˙ON and ONOO- were investigated. The data obtained show that TI generated a dose-dependent inhibition on production of nitrite and superoxide radicals. The IC50 value of TI on superoxide radical was 143.2 +- 4.29 .mu.g╱mL. And we used SOD activity staining to conform SOD activity of SPTI. SPTI also caused a dose-dependent inhibition of the oxidation of dihydrorhodamine 123 (DHR) by peroxynitrite. A calculated IC50 value of 809.1 +- 32.36 .mu.g╱mL was obtained on the inhibition of peroxynitrite radical. Spectrophotometric analyses revealed that TI suppressed the formation of ONOO--mediated tyrosine nitration through an electron donation mechanism. In further studies, TI also showed a significant ability of inhibiting nitration of bovine serum albumin (BSA) in a dose-dependent manner. In vivo TI inhibited LPS-induced nitrite production in macrophages in a concentration-dependent manner with an IC50 value of 932.8 +- 29.85 .mu.g╱mL. The present study suggested that TI had an efficient reactive nitrogen species scavenging ability. TI might be a potential effective NO and ONOO- scavenger useful for the prevention of the NO and ONOO- involved diseases.
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