NEW ASPECTS OF THE PROTECTIVES ON ATHEROSCLEROSIS BY FOODLIFE-REKATION WITH HDL-RECEPORS

HDL 受体的 FOODLIFE-REKATION 预防动脉粥样硬化的新方面

• 批准号: 09680049
• 负责人: HARA Takayuki
• 金额: $ 1.98万
• 依托单位: NAKAMURA GAKUEN UNUVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680049/
• 关键词: LIPOPROTEIN HIGH DENSITY; PROTECTION OF ATHEROSCLEROSIS; RECEPTORS; FOOD LIFE; 高密度リボ蛋白質

HIGH DENSITY LIPOPROTEIN (HDL) PLAYS AN IMPORTANT ROLE IN THE REVERSE CHOLESTEROL TRANSPORT FROM PERIPHERAL TISSUES TO LIVER TO REDUCE ACCUMULATING CHOLESTEROL IN THE ATHEROSCLEROTIC LEGIONS. HOWEVER, THE MECHANISM OF THE REVERSE CHOLESTEROL TRANSPORT BY HDL IS NOT FULLY UNDERSTOOD AND PARTICULARLY, THE INTERACTION BETWEEN HDL AND CELL SURFACE REMAINS TO BE ELUCIDATED. TO PROVE THE HYPOTHESIS THAT HDL RECEPTORS EXIST ON THE CELL SURFACE, SEVERAL HDL BINDNG PROTEINS HAVE BEEN DETECTED AND CHARACTERIZED IN PLASMA MEMBRANE FRACTIONS FROM VARIOUS TISSUES AND CELL LINES BY THE METHOD OF LIGAND BLOTTING. IN OUR PREVIOUS STUDY, WE REPORTED THAT TWO HDL-BINDNG PROTEINS (120 KDA AND 95 KDA) ARE DETCTED IN THE PLASMA MEMBRANE FRACTIONS OF BOVINE LIVER AND ADRENAL CORTEX USING A NEW LIGAND BLOTTNG METHOD USING APOLIPOPROTEIN AI (APOAI)-DNP AS A LIGAND. THEY ARE PROBABLY THE SAME AS HB1 AND HB2 THAT HAVE BEEN DETECTED AND PURIFIED BY FIDGE'S GROUP.IN THIS REPORT, WE PURIFIED A 95 KDA-HDL-BINDNG PROTEN FROM BOVINE LIVER BEING MOMITORED BY THE NEW LIGAND BLOTTING METHOD. WE REPORT THAT THE 95 KDA PROTEIN IS GR94 WHICH HAS BEEN KNOWN TO WORK AS A QUALITY CONTROL FOR SECRETORY PROTEINS. THE TRIAL OF LOOKING FOR A CANDIDATE FOR HDL-RECEPTOR WAS UNSUCCESSFUL AT THE MOMENT. HOWEVER, THIS FINDING PROVIDES THE FIRST EVIDENCE FOR DIRECT INTERACTION OF GRP94 WITH APO AI SINCE APO AI IS A SECRETORY PROTEIN SYNTHESIZED IN THE LIVER AND INTESTNE, AND IS BOUND TO GRP94 WITHOUT ANY AID OF CHEMICAL CROSS-LINKNG REAGENS.ON THE OTHER HAND, A l20 KDA HDL-BINDNG PROTEIN OF LIVER PLASMA MEMBRANE WAS FOUND TO INCREASE AFTER FEEDING OF CHOLESTEROL TO RATS OR RABBITS. THESE RESULTS SUGGEST THAT A 120 KDA HDL-BINDING PROTEIN MAY BE THE REAL CANDIDATE FOR HDL-RECEPTOR.

高密度脂蛋白（hdl）在从外周组织到肝脏的逆向胆固醇运输中起重要作用，以减少动脉粥样硬化军团中积累的胆固醇。然而，高密度脂蛋白胆固醇逆向转运的机制尚不完全清楚，特别是高密度脂蛋白与细胞表面的相互作用仍有待阐明。为了证明高密度脂蛋白受体存在于细胞表面的假设，用配体印迹法在不同组织和细胞系的质膜组分中检测和表征了几种高密度脂蛋白结合蛋白。在我们之前的研究中，我们报道了用载脂蛋白ai (apoai) -dnp作为配体的一种新的配体印迹方法，在牛肝脏和肾上腺皮质的质膜部分检测到两种hdl结合蛋白（120 kda和95 kda）。它们可能与菲奇小组已经检测和纯化的hb1和hb2相同。在本报告中，我们从牛肝脏中纯化了一种95 kda - hdl结合蛋白，并用新的配体印迹法进行了监测。我们报道了95 kda蛋白是gr94，它已经被认为是分泌蛋白的质量控制。寻找高密度脂蛋白受体候选体的试验目前尚未成功。然而，这一发现为grp94与载脂蛋白ai的直接相互作用提供了第一个证据，因为载脂蛋白ai是一种在肝脏和肠道中合成的分泌蛋白，并且不需要任何化学交联试剂的帮助就能与grp94结合。另一方面，大鼠或家兔饲喂胆固醇后，肝质膜高密度脂蛋白结合蛋白（hdl - binding PROTEIN）升高。这些结果表明，一个120 kda的hdl结合蛋白可能是hdl受体的真正候选蛋白。

项目成果

楠 喜久枝、藤田 守、松隈美紀、馬場良子、三成由美、田所忠弘、印南 敏、前川昭男: "カキの葉の組織構造並びに食物繊維成分について" 日本食品保蔵科学会誌. Vol.23(2). 65-75 (1997)

Kikue Kusunoki、Mamoru Fujita、Miki Matsukuma、Ryoko Baba、Yumi Mitsunari、Tadokoro Tadokoro、Satoshi Inami、Akio Maekawa：“关于柿子叶的组织结构和膳食纤维成分”日本食品保存科学学会杂志第 23 卷。 (2). 65-75 (1997)。

Fujita,M., Baba,R., Matsuguma,M., Kai,T., Nozaki,S., Miyoshi,M.: "Apical and basolateral endocytic pathways in absorptive cells of suckling rat adult rats small intestine in vivo" Digestion & Absorption. Vol.20(2). 113-118 (1997)

Fujita,M.、Baba,R.、Matsuguma,M.、Kai,T.、Nozaki,S.、Miyoshi,M.：“体内乳鼠成年大鼠小肠吸收细胞的顶端和基底外侧内吞途径”消化

Baba,R., Matsuguma,M., Fujita,M., Kai,T., Nozaki,S., Miyoshi,M.: "Endocytosis by absorptive cells of the adult rats large intestine in vivo" Digestion & Absorption. Vol.20(2). 129-132 (1997)

Baba,R.、Matsuguma,M.、Fujita,M.、Kai,T.、Nozaki,S.、Miyoshi,M.：“体内成年大鼠大肠吸收细胞的内吞作用”消化