Regulatory mechanism of aquaporin expression in cerebral microvessels : its pathophysiological significance

脑微血管水通道蛋白表达的调控机制及其病理生理意义

• 批准号: 13672394
• 负责人: KOBAYASHI Hideyuki
• 金额: $ 2.18万
• 依托单位: 宮崎医科大学
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672394/
• 关键词: Cerebral microvessel; Aquaporin; Cultured endothelial cell; Ischemia; hypoxia; Astrocytes; Regulation of expression; Cytotoxic edema; Immortalized cell; RT-PCR; ウエスタンブロット; 培養; 脳浮腫

To clarify the regulatory mechanism of water transport between blood and brain, we have identified water channel aquaporin (AQP) subtypes expressed in the rat cerebral microvessels. mRNA and protein of AQP1 and AQP4 were present in the microvessels prepared from rat cerebral cortex. AQP1 mRNA was expressed abundantly as kidney but the AQP1 mRNA level was 1/100 of the kidney. AQP4 protein was detected as 30 kDa band with higher molecular weight bands consisted of its oligomer and smear band hardly solubilized form by western blot AQP1 was detected as a 28 kDa band and its level was less than 1/10 of that in the kidney. AQP4 was detected at the cell membrane of the microvessel, but AQP1 protein was hardly detectable by immunohistochemistry. The expression of AQP1 was decreased by the addition of conditioned medium of astrocytes in the primary culture of the microvessels as well as in the immortalized cell line of the rat microvessels, GP8. The expression of AQP1 increased under hypoxic condition.The regulation of AQP1 expression by astrocyte-derived factor and its increase by hypoxia suggest that AQP1 is a key molecule involved in the generation of cytotoxic edema.

为了阐明血脑水运的调控机制，我们鉴定了大鼠脑微血管中表达的水通道水通道蛋白（AQP）亚型。在大鼠大脑皮层制备的微血管中存在AQP1和AQP4的mRNA和蛋白。肾脏AQP1 mRNA表达丰富，但AQP1 mRNA水平为肾脏的1/100。AQP4蛋白为30 kDa条带，较高分子量条带由其寡聚物和难以溶解的涂片条带组成，western blot检测到AQP1蛋白为28 kDa条带，其水平不到肾脏的1/10。在微血管细胞膜上检测到AQP4蛋白，但免疫组化几乎检测不到AQP1蛋白。在微血管原代培养和大鼠微血管永生化细胞系GP8中加入星形胶质细胞条件培养基可降低AQP1的表达。缺氧条件下AQP1表达增加。星形胶质细胞衍生因子对AQP1表达的调控以及缺氧对AQP1表达的增加提示AQP1是参与细胞毒性水肿发生的关键分子。

项目成果

Kobayashi H., Minami S., Itoh S., Shiraishi S., Yokoo H., Yanagita T., Uezono Y., Mohri M. and Wada A.: "Aquaporin subtypes in rat cerebral microvessels."Neuroscience Letters. 297. 163-166 (2001)

Kobayashi H.、Minami S.、Itoh S.、Shiraishi S.、Yokoo H.、Yanagita T.、Uezono Y.、Mohri M. 和 Wada A.：“大鼠脑微血管中的水通道蛋白亚型。”神经科学快报。

Kis B., Kaiya H., Nishi R., Deli M. A., Abraham C. S., Yanagita T., Isse T., Gotoh S., Kobayashi H., Wada A., Niwa M., Kangawa K., Greenwood J., Yamashita H. and Ueta Y.: "Cerebral endothelial cells are a major source of adrenomedullin."Journal of Neuroen

Kis B.、Kaiya H.、Nishi R.、Deli M.A.、Abraham C.S.、Yanagita T.、Isse T.、Gotoh S.、Kobayashi H.、Wada A.、Niwa M.、Kangawa K.、Greenwood J.、

Kobayashi H., Yokoo H., Yanagita T., and Wada A.: "Regulation of cerebral microvessel function : Innervation and bioactive peptides"Japanese Journal of Circulation Research. 25. 155-164 (2002)

Kobayashi H.、Yokoo H.、Yanagita T. 和 Wada A.：“脑微血管功能的调节：神经支配和生物活性肽”日本循环研究杂志。

B.Kis: "Adrenomedullin, an autocrine mediator of the blood-brain barrier function"Hypertension Research. 26. s61-s70 (2003)

B.Kis：“肾上腺髓质素，血脑屏障功能的自分泌介质”高血压研究。

Kobayashi H., Yokoo H., Yanagita T., and Wada A.: "Regulation of cerebral microvessel function"Folia Pharmacologica Japonica. 119. 281-286 (2002)

小林H.、横尾H.、柳田T.、和田A.：“脑微血管功能的调节”Folia Pharmacologica Japonica。