Modulation of ectoenzyme-gene expression in volving reformation of bile-canalicular structures in rat primary cultured hepatocytes

大鼠原代培养肝细胞胆小管结构重建中外酶基因表达的调节

• 批准号: 02670304
• 负责人: IKAWA Shiro
• 金额: $ 1.41万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02670304/
• 关键词: Primary cultured of rat hepatocytes; Bile canalicular reformation; Cell membrane polarity; Bile canalicular ectoenzyme; Taurocholate binding assay; Bile acid carrier protein; Labeling of bile acid carrier protein; 胆汁酸結合蛋白質; 細胞極性; 標識結合蛋白質の分布; アクチ

The isolated hepatocytes membranes and bile-canalicular structures are damaged by collagenase digestion. These damages, however, are almost completely repaired when the hepatocytes are cultured as monolayers for only 24hr. Moreover, we found that the suppressive effects of insulin on bile-canalicule reformation or alkaline phosphatase induction by dexamethasone were due to decrease and the suppression of synthesis of alkaline mRNA, not to enhacement of its degradation. However, the relation to this enzyme induction is still unknown. Therefore, further studies are necessary for the regulatory gene of this enzyme and the characterization of the transmitter molecules and/or effector molecules involved in transcription and their interaction with genetic structure. On the other hand, the bile acid binding protein checked by [^<14>C]taurocholate is distributed mainly on the surface of the bile canalicule. This binding protein from rat liver plasma membrane was fractionated by Sephacryl S-200 column ; molecular weight is 100kD, 60kD and 48kD, respectively. This binding protrin (100kD) was mainly found from sinusoidsite. This is in accordance with intracellular for bile acid transport.

分离的肝细胞膜和胆管结构被胶原酶消化破坏。然而，当肝细胞单层培养仅24小时时，这些损伤几乎完全修复。此外，我们还发现胰岛素对地塞米松诱导的胆管重构或碱性磷酸酶的抑制作用是由于减少和抑制碱性mRNA的合成，而不是促进其降解。然而，这种酶诱导的关系仍然未知。因此，有必要对该酶的调控基因以及参与转录的传递分子和/或效应分子的表征及其与遗传结构的相互作用进行进一步的研究。另一方面，由[^ C]牛磺胆酸盐检查的胆汁酸结合蛋白<14>主要分布在胆小管的表面。用Sephacryl S-200柱从大鼠肝细胞膜上分离得到的结合蛋白分子量分别为100 kD、60 kD和48 kD。这种结合蛋白（100 kD）主要存在于窦状隙中。这与胆汁酸的细胞内转运是一致的。

项目成果

武良 哲雄: "ラット初代培養肝細胞における微小胆管再形成に与えるホルモン添加の影響"

Tetsuo Takera：“激素添加对原代培养大鼠肝细胞微胆管重塑的影响”

K.SATO: "Negative Regulatio of Catalase Gene Expression in Hepatoma Cells" Molecular and Cellular Biology. 12. 2525-2533 (1992)

K.SATO：“肝癌细胞中过氧化氢酶基因表达的负调节”分子和细胞生物学。

K.Sato: "Negative Regulatio of Catalase Gene Expression in Hepatoma Cells" Molecular and Cellular. 12. 2525-2533 (1992)

K.Sato：“肝癌细胞中过氧化氢酶基因表达的负调节”《分子与细胞》。

Y Kohno: "The Effect of Human Milk on DNA Synthesis of Neonatal Rat Hepatocytes in Primary Culture" Pediatric Research. 29. 251-255 (1991)

Y Kohno：“母乳对原代培养中新生大鼠肝细胞 DNA 合成的影响”儿科研究。

武良 哲雄: "Purification and Properties of Bile Acid Carrier Protein from Rat Liver Plasma Membranes"

Tetsuo Takera：“大鼠肝质膜胆汁酸载体蛋白的纯化和性质”