Preparation of Polymer-Conjugated Bilirubin Oxidase and Treatment of Jaundice

聚合物结合胆红素氧化酶的制备及治疗黄疸

• 批准号: 01570164
• 负责人: MAEDA Hiroshi
• 金额: $ 1.28万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570164/
• 关键词: Influenza virus; Pathogenesis; Active oxygens; Free radical; Superoxide dismutase; Radical scavenger; Chronic viral hepatitis; Chronic viral diseases; 活性酵素; プロテア-ゼ; 複合感染

The purpose of the present investigation is to clarify the role of bacterial protease in the influenza infection and the detailed analysis pathogenic process in influenza infection. The protease used was obtained from Serratia marcescens as a model of bacterial infection. When mice were infected witt influenza virus, addition of the protease yielded remarkable increase (about 100 fold) of virus in the lung of the infected mice compared with those without protease. This was found as a synergistic outcome in the survival of mice ; all mice died at much accelerated rate, where about 40% died without the protease. Next question is that what is the pathogenic molecules in influenza infection ; virus itself or other indirect consequence. We focused free radical generation in the Bronchiol Alveolar Lavage Fluid (BALF). To our surprise the level of xanthiee oxidase (OX) was elevated 400 fold higher than that of control. XO is an enzyme which generate O_2^- as well as OH radicals using hypoxant … More hine or xanthine as its substrate. The level of NADPH oxidase, another source of O_2^- generation was elevated at most 7 fold in BALF. Then, we investigated upstream cascadeof hypoxanthine or xanthine generation ; which is inosine derived after deamination of adenosine by adenosine deaminase.The adenosine deaminase activity was elevated 20 fold of uninfected mice ; a solid evidence of enhanced supply of fuel for XO. Then, we tested effect of removal of O_2^- cure 90% of virus infected mice otherwise all died. This finding was further strengthened by the fact that inhibition of XO by allopurinol was beneficial to the survival of mice. Generation of XO from xanthine dehydrogenase is mediated via serine protease which can be inhibited by soybean trypsin inhibitor. The effect of this inhibitor was found also effective to the viral infected mice for their survival. This inhibitor may be effective also inhibiting activation of virus by serine protease in BALF and also inhibiting kallikrein which generate bradykinin a major cause of vascular permeability and inflammation. Less

本研究的目的是阐明细菌蛋白酶在流感感染中的作用，并详细分析流感感染的致病过程。所使用的蛋白酶是从粘质沙雷氏菌中获得的，作为细菌感染的模型。当小鼠感染Witt流感病毒时，加入该酶可使感染小鼠肺内病毒含量显著增加(约为未加酶小鼠的100倍)。这被发现是小鼠存活的协同结果；所有小鼠的死亡速度都大大加快，其中约40%的小鼠在没有蛋白酶的情况下死亡。下一个问题是，流感感染中的致病分子是什么；病毒本身还是其他间接后果。我们重点研究了支气管肺泡灌洗液中自由基的产生。令我们惊讶的是，黄色素氧化酶(OX)的水平比对照组高出400倍。XO是一种利用低氧…产生O2^-和OH自由基的酶更多的是以黄嘌呤或黄嘌呤为底物。BALF中O_2~-产生的另一来源NADPH氧化酶水平最高可提高7倍。然后，我们研究了次黄嘌呤或黄嘌呤的上游级联；次黄嘌呤是腺苷脱氨酶脱氨后产生的肌苷。腺苷脱氨酶活性在未感染的小鼠中提高了20倍；这是XO燃料供应增加的坚实证据。然后对感染病毒的小鼠进行脱氧效果检测，90%的小鼠全部死亡。别嘌醇抑制XO有利于小鼠的存活，这一事实进一步加强了这一发现。黄嘌呤脱氢酶通过丝氨酸蛋白酶产生XO，丝氨酸蛋白酶可被大豆胰蛋白酶抑制剂抑制。该抑制物对病毒感染小鼠的存活也有效。这种抑制剂可能还能有效地抑制BALF中丝氨酸蛋白酶对病毒的激活，并抑制激肽释放酶，激肽释放酶是导致血管通透性和炎症的主要原因。较少

项目成果

A.Molla,T.Akaike,and H.Maeda: "Inactivation of various proteinase inhibitors and the complement system in human plasma by the 56ーkilodalton proteinase from serratia marcescens." Infect.Immun.57. 1868-1871 (1989)

A. Molla、T. Akaike 和 H. Maeda：“来自 Infect.Immun.57 的 56 千道尔顿蛋白酶对人血浆中的各种蛋白酶抑制剂和补体系统的灭活”。

Akhteruzzaman Molla: "Inactivation of various proteinase inhibitors and the complement system in human plasma by the 56kilodalton proteinase from Serratia Marcescens." Infect. Immun.57. 1868-1871 (1989)

Akhteruzzaman Molla：“来自粘质沙雷氏菌的 56 千道尔顿蛋白酶可灭活人血浆中的各种蛋白酶抑制剂和补体系统。”

Takaki Akaike: "Dependence on O_2^- generation by xanthine oxidase of pathogenesis of influenza virus infection in mice." J. Clin. Invest.85. 739-745 (1990)

Takaki Akaike：“小鼠流感病毒感染发病机制对黄嘌呤氧化酶产生 O_2^- 的依赖性。”

Yasuhiro Matsumura: "Degradation pathway of kinins in tumor ascites and inhibition by kininase inhibitors : Analysis by HPLC." Agents and Actions.29. 171-180 (1990)

Yasuhiro Matsumura：“肿瘤腹水中激肽的降解途径和激肽酶抑制剂的抑制：HPLC 分析。”

赤池 孝章,吉岡 誠,前田 浩,小田 達也,鈴木 富士夫,安藤 正幸,荒木 淑郎,: "マウスインフルエンザウイルス感染症における病因としての活性酸素の役割と活性酸素産生カスケ-ドの亢進" 医学のあゆみ. 148. 829-830 (1989)

Takaaki Akaike、Makoto Yoshioka、Hiroshi Maeda、Tatsuya Oda、Fujio Suzuki、Masayuki Ando、Shukuro Araki，：“活性氧作为鼠流感病毒感染原因的作用以及活性氧产生级联的增强”的历史医学148.829-830（1989）