Contribution of glutathione to multidrug resistance and the its regulation with active oxygen modulators

谷胱甘肽对多药耐药性的贡献及其活性氧调节剂的调节

• 批准号: 08672521
• 负责人: ADACHI Tetsuo
• 金额: $ 0.77万
• 依托单位: Gifu Pharmaceutical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672521/
• 关键词: Glutathione; Multidrug resistance-associated protein; Active oxygen; Superoxide dismutase; Glucuronosyl conjugate

Multidrug resistance-associated protein (MRP) has shown to transport glutathione S-conjugate. The effect of glutathione (GSH) on the transport of glucuronosyl etoposide, a major metabolite of etoposide in MRP overexpressing cell line KB/VP-4.1.The involvement of MRP in the transport of drug conjugates was investigated using membrane vesicles prepared from KB/VP-4 cells. The ATP-dependent transport of not only GSH-conjugate such as [^3H]-leukotriene C4, but also glucuronosyl conjugate of [^3H] etoposide. This result demonstrates that MRP is a pump with a broad specificity.2.The butionine sulfoximine (BSO)-exposure reduced the GSH level inKB/VP-4 cells. The efflux of glucuronosyl etoposide was5-times faster from KB/VP-4 than from KB cells (wild type cells). BSO abolished the increased efflux of drug from KB/VP-4.3.In KB/VP-4 cells, the contents of copper, zinc-superoxide dismutase and manganese-superoxide dismutase were 1.5-fold and 6-fold higher than those in KB cells, respectively. This result shows that KB/VP-4 cell might to be more resistant to active oxygens.4.Molecular genetic studies of extracellular-superoxide dismutase have shown that a single base substitution generating the exchange of glycine for arginine 213 causes the impairment of its binding ability to endothelial cell surface and susceptibility to trypsin-like proteinases. The binding of EC-SOD to cell surface is an especially efficient way of protecting cells against external superoxide anion. The decrease of the protective capacity on the endothelial surface by EC-SOD might accelerate the renal, cardiovascular dysfunctions, diabetes and hyperlipidemia.The change of SOD isozyme might affect on the ATP-dependent export of drug which are controlled by active oxygen modulators such as GSH.

多药耐药相关蛋白可转运谷胱甘肽S结合物。谷胱甘肽(GSH)对足叶乙甙在MRP高表达细胞系KB/VP-4.1中的主要代谢产物葡萄糖醛酸依托泊苷转运的影响。依赖于ATP的转运不仅包括GSH结合物如[~3H]-白三烯C4，还包括[~(3 H)]依托泊苷的葡萄糖醛酸基结合物。这一结果表明MRP是一个具有广泛特异性的泵。2.丁氨酸亚磺胺(BSO)暴露降低了KB/VP-4细胞的GSH水平。在KB/VP-4细胞中，葡萄糖醛酸依托泊苷的外流速度是KB细胞(野生型细胞)的5倍。在KB/VP-4细胞中，铜、锌超氧化物歧化酶和锰超氧化物歧化酶的含量分别是KB细胞的1.5倍和6倍。这一结果表明KB/VP-4细胞可能对活性氧具有更强的抵抗力。4.细胞外超氧化物歧化酶的分子遗传学研究表明，单碱基取代导致甘氨酸与精氨酸213的交换导致其与内皮细胞表面的结合能力受损，并对类胰蛋白酶敏感。EC-SOD与细胞表面的结合是保护细胞免受外界超氧阴离子伤害的一种特别有效的方法。EC-SOD对血管内皮细胞表面保护能力的降低可能会加速肾脏、心血管功能障碍、糖尿病和高脂血症的发生；SOD同工酶的变化可能会影响受GSH等活性氧调节剂调控的ATP依赖的药物输出。

项目成果

Harutaka Yamada: "Polymorphism of extracellular superoxide dismutase (EC-SOD) gene:Relation to the mutation responsible for high EC-SOD level in serum" Japanese Journal of Human Genetics. 42. 353-356 (1997)

Harutaka Yamada：“细胞外超氧化物歧化酶 (EC-SOD) 基因的多态性：与血清中高 EC-SOD 水平的突变的关系”《日本人类遗传学杂志》。

Harutaka Yamada, Mitsuru Kitano, Atsushi Fukatsu, Mintan Son, Arao Futenma, Shinichi Kakumu, Yasukazu Yamada, Tetsuo Adachi, Kazuyuki Hirano: "Moleccular analysis of extracellular-superoxide dismutase in patients with chronic renal failure." Kidney and Fr

Harutaka Yamada、Mitsuru Kitano、Atsushi Fukatsu、Mintan Son、Arao Futenma、Shinichi Kakumu、Yasukazu Yamada、Tetsuo Adachi、Kazuyuki Hirano：“慢性肾功能衰竭患者细胞外超氧化物歧化酶的分子分析。”

Tetsuo Adachi: "An arginine-213 to glycine mutation in human extracellular-superoxide dismutase reduces susceptibility to trypsin-like proteinases" Journal of Biochemistry. 120. 184-188 (1996)

Tetsuo Adachi：“人细胞外超氧化物歧化酶中精氨酸 213 向甘氨酸的突变降低了对胰蛋白酶样蛋白酶的敏感性”《生物化学杂志》。

Tetsuo Adachi: "An arginine-213 to glycyine mutation in human extracellular-superoxide dismutase reduces susceptibility to trypsin-like proteinases" Journal of Biochemistry. 120. 184-188 (1996)

Tetsuo Adachi：“人细胞外超氧化物歧化酶中精氨酸 213 向甘氨酸的突变降低了对胰蛋白酶样蛋白酶的敏感性”《生物化学杂志》。

北野 充: "腎とフリーラジカル" 加藤 克己,長瀬 光昌, 3 (1997)

北野充：《肾脏与自由基》加藤克己、长濑光政，3 (1997)