Membrane damage model of lipid peroxidation, using carboxyfluorescein enveloping liposome

使用羧基荧光素包封脂质体的脂质过氧化膜损伤模型

• 批准号: 04670208
• 负责人: OKADA Shigeru
• 金额: $ 1.34万
• 依托单位: Okayama University Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670208/
• 关键词: carboxyfluorescein; liposome; lipid peroxidation; polyunsaturated fatty acid; saturated fatty acid; ferric nitrilotriacetate; 鉄ニトリロ三酢酸; 酸化還元活性; 脂質過酸化; 遷移金属; Fenton様反応; 共役ジエン; 鉄キレート

A Carboxyfluorescein (CF) -enveloping liposome is presented as a physicochemical damage model of the biomembrane for the study of lipid peroxidation. The degree of membrane damage may be assessed by measurement of the fluorescence intensity of CF released from damaged liposomes after the interaction between the liposomal membrane and a radical initiator. The process of lipid peroxidation was biochemically confirmed by the thiobarbituric acidreactive substances (TBARS) test done simultaneously. Three types of CF-enveloping liposomes, i.e., soybean phosphatidylcholine (SPC) containing polyunsaturated fatty acid, dipalmitoylphosphatidylcholine (DPPC) containing saturated fatty acid, and soybean phosphatidylcholine plus alpha-tocopherol, were exposed to a known radical initiator, ferric nitrilotriacetate with a reductant (either cysteine, reduced form glutathione, or ascorbic acid). The release of CF from the peroxidized liposome was measured with a fluorospectrophotometer. A high release of CF and a high level of TBARS were seen in the SPC liposomes, but not in the DPPC and alpha-tocopherol-inserted SPC liposomes. This liposome model is superior to the conventional red cell hemolysis model for several reasons : (1) the constituents of the membrane are defined ; (2) CF is not a radical initiator, whereas hemoglobin is a potential radical initiator ; and (3) the membrane components can be easily changed depending on the purpose of the experiment.

为了研究脂质过氧化作用，提出了一种包裹羧基荧光素(CF)的脂质体作为生物膜的物理化学损伤模型。膜损伤的程度可以通过测量脂膜与自由基引发剂相互作用后从受损脂质体释放的CF的荧光强度来评估。同时进行硫代巴比妥酸反应物质(TBARS)检测，以生化方法证实脂质过氧化过程。将含多不饱和脂肪酸的大豆磷脂酰胆碱(SPC)、含饱和脂肪酸的二棕榈酰磷脂酰胆碱(DPPC)和大豆磷脂酰胆碱+α-生育酚三种类型的CF包膜脂质体暴露于已知的自由基引发剂--含有还原剂(半胱氨酸、还原型谷胱甘肽或抗坏血酸)的氮三乙酸铁。用荧光分光光度计法测定过氧化脂质体释放量。SPC脂质体有较高的CF释放量和较高的TBARS释放量，而DPPC脂质体和α-生育酚SPC脂质体则无此现象。这种脂质体模型优于传统的红细胞溶血模型，原因如下：(1)定义了膜的组成；(2)CF不是自由基引发剂，而血红蛋白是潜在的自由基引发剂；(3)根据实验目的，膜成分可以很容易地改变。

项目成果

Ying-yan Yu, Tetsuya Ogino, Shigeru Okada: "Histochemical detection of lipid peroxidation in human gastrointestinal, mammary and renal carcinomas." Acta Med.Okayama. 46. 233-239 (1992)

Ying-yan Yu、T​​etsuya Ogino、Shigeru Okada：“人类胃肠癌、乳腺癌和肾癌中脂质过氧化的组织化学检测。”

Daxian Zhang, Tatsuji Yasuda, Shigeru Okada: "A carboxyfluorescein - enveloping liposome as a physicochemical damage model of the biomembrane for the study of lipid peroxidation." J.Clin.Biochem.Nutr.14. 83-90 (1993)

Daxian Zhang、Tatsuji Yasuda、Shigeru Okada：“一种羧基荧光素 - 包封脂质体作为生物膜的理化损伤模型，用于研究脂质过氧化。”

Teruyuki Kawabata, Shigeru Okada, Miao Liu, Akira Asaumi, Takashi Akiyama: "Action of pyridinecarboxamides on Fe(III)-NTA induced renal toxicity and superoxide anion." Oxygen Radicals.Ed.K.Yagi, M.Kondo, E.Niki and T.Yoshikawa, Elsevier Science Publishers

Teruyuki Kawabata、Shigeru Okada、Miao Liu、Akira Asaumi、Takashi Akiyama：“吡啶甲酰胺对 Fe(III)-NTA 诱导的肾毒性和超氧阴离子的作用。”

T.Kawabata,S.Okada, T.Akiyama,A.Asaumi: "Generation of Superoxide Anion in Fe(III)-nta(nta=nitrilotriacetate) and H_2O_2 System. An ESR Study." Chemistry Letters. 2189-2192 (1992)

T.Kawabata,S.Okada, T.Akiyama,A.Asaumi：“Fe(III)-nta(nta=次氮基三乙酸盐) 和 H_2O_2 系统中超氧阴离子的生成。ESR 研究。”

Daxian Zhang: "A carboxyfluorescein-enveloping liposome as a physicochemical damage model of the biomembrane for the study of lipid peroxidation." J.Clin.Biochem.Nutr.14. 83-90 (1993)

张大贤：“一种羧基荧光素包封的脂质体作为生物膜的理化损伤模型，用于研究脂质过氧化。”