Production and application of novel model animals with an oxidative stress

新型氧化应激模型动物的制备及应用

• 批准号: 08557014
• 负责人: ARAI Hiroyuki
• 金额: $ 9.54万
• 依托单位: GRADUATE SCHOOL OF PHARMACEUTICAL SCIENCES,THE UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08557014/
• 关键词: VITAMINE; OXIDATIVE STRESS; KNOCKOUT MOUSE; BERGMANN GLIA; 小脳; プルキンエ細胞; バーグマングリア細胞; 輸送蛋白質; 基質特異性

Vitamin E (a-tocopherol) is a fat-soluble antioxidant that is transported by plasma lipoproteins in the body. a-tocopherol taken up by the liver with lipoprotein is thought to be re-secreted into the plasma in very low density lipoprotein (VLDL), alpha-tocopherol transfer protein (alphaTTP), which was recently identified as a product of the causative gene for familial isolated vitamin E deficiency, is a cytosolic liver protein and plays an important role in the efficient recycling of plasma vitamin E.Using cell culture system, we found that the secretion of alpha-tocopherol is more efficient in cells expressing alphaTTP than in matched cells lacking alphaTTP.Brefeldin A, which inhibits VLDL secretion by disrupting the Golgi apparatus, has no effect on alpha-tocopherol secretion, indicating that alphaTTP-mediated alpha-tocopherol secretion is not coupled to VLDL secretion. Among other agents tested, only 25-hydroxycholesterol, a modulator of cholesterol metabolism, inhibits alpha-tocoph … More erol secretion. These results suggest that alphaTTP functions to stimulate secretion of cellular alpha-tocopherol into the extracellular medium and that the reaction utilizes a novel non-Golgi mediated pathway which may be linked to cellular cholesterol metabolism or transport. We examined the structural characteristics of vitamin E analogs required for recognition by alphaTTP.Interestingly, there is a linear relationship between the relative affinity and the known biological activity obtained from the rat resorption-gestation assay. Thus the affinity of vitamin E analogs for alphaTTP is one of the critical determinants of their biological activity. alphaTTP is exclusively expressed in the liver, but the message for alphaTTP was detectable at low levels in some rat tissues including brain, spleen, lung and kidney. In the brain, the alphaTTP transcript is detected predominantly in the cerebellar cortex, as revealed by in situ hybridization histochemistry. In the celebellar cortex, the hybridization signals are detected mostly in the Bergmann glial cells. We have recently succeeded in establishing the knock out mice for alphaTTP.The plasma vitamin E level of the mice is almost undetectable and that of heterozygote is about half that of wild type mice. These data clearly demonstrate that alphaTTP is a major determinant of plasma vitamin E level. Less

维生素E(α-生育酚)是一种脂溶抗氧化剂，由体内的血浆脂蛋白运输。肝脏摄取的α-生育酚与脂蛋白一起被认为以极低密度脂蛋白(VLDL)的形式重新分泌到血浆中。α-生育酚转移蛋白(AlphaTTP)是一种细胞质的肝脏蛋白，在血浆维生素E的有效循环中起着重要作用。通过细胞培养系统，我们发现在表达α-TTP的细胞中，α-生育酚的分泌比缺乏α-TTP的细胞更有效。Brefeldin A通过干扰高尔基器来抑制VLDL的分泌，但对α-生育酚的分泌没有影响。提示α-TTP介导的α-生育酚的分泌与极低密度脂蛋白的分泌没有偶联。在其他被测试的药物中，只有25-羟基胆固醇，一种胆固醇新陈代谢的调节剂，抑制α-生育蛋白…更多的性爱分泌。这些结果表明，α-TTP具有刺激细胞α-生育酚分泌到细胞外介质的功能，该反应利用了一种新的非高尔基体介导的途径，该途径可能与细胞胆固醇的代谢或运输有关。我们研究了α-TP识别所需的维生素E类似物的结构特征。有趣的是，相对亲和力与从大鼠吸收-妊娠试验中获得的已知生物活性之间存在线性关系。因此，维生素E类似物对α-TTP的亲和力是其生物活性的关键决定因素之一。AlphaTTP仅在肝脏中表达，但在一些大鼠组织中可检测到低水平的AlphaTTP信息，包括脑、脾、肺和肾脏。在大脑中，正如原位杂交组织化学显示的那样，AlphaTTP转录本主要在小脑皮质中检测到。在人脑皮质，杂交信号主要在Bergmann神经胶质细胞中检测到。我们最近成功地建立了αTTP基因敲除小鼠，其血浆维生素E水平几乎检测不到，杂合子小鼠的血浆维生素E水平约为野生型小鼠的一半。这些数据清楚地表明，αTTP是血浆维生素E水平的主要决定因素。较少

项目成果

Matsuzawa Atsushi et al.: "Protection against oxidative stress-induced cell death by intracellular platelet-activating factor-acetylhydrolase II" Journal of Biological Chemistry. 272巻. 32315-32320 (1997)

Matsuzawa Atsushi 等人：“细胞内血小板激活因子乙酰水解酶 II 防止氧化应激诱导的细胞死亡”，生物化学杂志，第 272 卷，32315-32320（1997 年）。

T.Yokota, T.Shiojiri, T.Gotoda and H.Arai: "Retinitis pigmentosa and ataxia caused by a mutation in the gene for the alpha-tocopherol-transfer protein" New Engl.J.Med.335. 1770 (1996)

T.Yokota、T.Shiojiri、T.Gotoda 和 H.Arai：“α-生育酚转移蛋白基因突变引起的色素性视网膜炎和共济失调”New Engl.J.Med.335。

Yokota T.et al.: "Retinitis Pigmentosa and Ataxia Caused by a Mutation in the Gene for the α-Tocopherol Transfer Protein" N.Engl.J.Med.335. 1770-1771 (1996)

Yokota T.等人：“α-生育酚转移蛋白基因突变引起的视网膜色素变性和共济失调”N.Engl.J.Med.335（1996）。

Arita,M.et al.: "Binding of α-tocopherylquinone,an oxidized from of δ-tocopherol toglutathione-S-transferase in the liver cytosol" FEBS Letters. 436. 424-426 (1998)

Arita, M. 等人：“α-生育酚醌（肝细胞质中 δ-生育酚谷胱甘肽-S-转移酶的氧化形式）的结合”FEBS Letters 436. 424-426 (1998)。

Matzuzawa,A.et al.: "Protection against oxidative stress-induced cell death by intracellular platelet-activating factor-acetylhydrolase II" Journal of Biological Chemistry. 272. 32315-32320 (1997)

Matzazawa,A.et al.：“细胞内血小板激活因子乙酰水解酶 II 防止氧化应激诱导的细胞死亡”《生物化学杂志》。