Studies on Pathogenesis of Intrahepatic Cholestasis - Analysis of Ca^<++>- Calmodulin-Actomyosin System.

肝内胆汁淤积发病机制的研究-Ca^<>-钙调蛋白-肌动球蛋白系统分析。

• 批准号: 62570335
• 负责人: ODA Masaya
• 金额: $ 1.6万
• 依托单位: Keio 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: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570335/
• 关键词: intrahepatic cholestasis; canalicular contraction; actin filaments; intracytoplasmec Ca^<++>; calmodulin; primary biliary cirrhosis; anticytokeratin antibody; HLA-DR; Calcium; calmodulin; antin; 胆汁うっ滞; fura2; calcium antimonate tetroxide法; 抗

In an attempt to elucidate the pathogenesis of intrahepatic cholestasis due to bile secretory impairment, the cytoskeletal system in the intrahepatic bilary tract was investigated in vivo and in vitro by immunofluorescence and immunoelectron microscopy using an experimental model of acute intrahepatic cholestasis and liver biopsy specimens obtained from patients with acute intrahepatic cholestasis. The ultrastructural distribution of intracytoplamic free calcium ions (Ca^<++>) and its alterations were revealed by digital image analysis of the electron micrographs obtained by the potassium antimonate method. Bile canalicular contractions were analysed by time-lapse cinematography. It has been probed that the existence of intracytoplasmic Ca^<++>, Ca^<++>-binding protein calmodulin, and Ca^<++>-Mg^<++>-ATPase is essential for the control of actin filaments-mediated bile canalicular contractions. The depletion of these three factors resulted in the disturbances of bile canalicular contrac … More tions, i. e. the paralytic dilatation of bile canaliculli, causing stagnation of bile in the canaliclus network leading to intrahepatic cholestasis.Anti-intermediate filaments (IF) and anti-actin filaments (AF) were determined in sera of patients with primary biliary cirrhosis (PBC) by immunofluorescence and immunoperoxidase method using the monolayer-culutured cell lines, _3T_3 and PtK_2. It has been considered that anti-IF antibody is formed against the IF derived from the bile duct epithelium as a result of bile duct destruction, while anti-IF antibody is formed mainly against the AF derived from the luminal site of bile duct epithelium at stage I of PBC and is further formed against the AF from pericanalicular region after the stage II. Immunofluorescence and immunoperoxidase microscopy revealed the aberrant expression of HLA-DR within the intrahepatic bile duct epithelium in the biopsy speciments from patients with PBC. It is strongly suggested that cytotoxic T cells may recognize this HLA-DR and react with the bile duct epithelium, leading to the destruction of bile duct in PBC. Less

为了阐明胆汁分泌障碍所致肝内胆汁淤积的发病机制，采用急性肝内胆汁淤积的实验模型和急性肝内胆汁淤积患者的肝活检标本，通过免疫荧光和免疫电镜在体内和体外研究了肝内胆道的细胞骨架系统。用锑酸钾法观察细胞质内游离钙离子（Ca^++）的分布及其变化。通过延时电影摄影分析胆小管收缩。已经探索了胞浆内Ca^++、Ca^++结合蛋白钙调素和Ca^++-Mg^++-ATP酶的存在对于控制肌动蛋白介导的胆小管收缩是必需的。这三种因子的缺失导致了胆管收缩功能的紊乱。 ...更多信息 蒂申岛e.本文应用免疫荧光和免疫过氧化物酶法检测了原发性胆汁性肝硬化（PBC）患者血清中抗中间纤维（IF）和抗肌动蛋白纤维（AF）。已经认为，抗IF抗体是针对由于胆管破坏而来自胆管上皮的IF而形成的，而抗IF抗体主要是针对PBC的I期时来自胆管上皮的管腔部位的AF而形成的，并且在II期之后进一步针对来自小管周围区域的AF而形成。免疫荧光和免疫过氧化物酶显微镜显示PBC患者肝内胆管上皮细胞HLA-DR异常表达。提示细胞毒性T细胞可能识别该HLA-DR抗原，并与胆管上皮细胞发生反应，导致PBC时胆管的破坏。少

项目成果

Masaya Oda: "Intrahepatic distribution of laminin and its producing cells in carbon tetrachlorideーinduced liver cirrhosisーAn immunoーfluorerescense and immunoelectron microscopic studyー." J.Clin.Electron Microscopy. 20. 560-561 (1987)

Masaya Oda：“四氯化碳诱导的肝硬化中层粘连蛋白及其产生细胞的肝内分布 - 免疫荧光和免疫电子显微镜研究。”J.Clin.Electron Microscopy 20. 560-561 (1987)。

Watanabe, N., Miyairi, M., Suyama, K., Oshio, C., Oda, M. and Tsuchiya, M.: "Effects of taurolithocholate on the bile canalicular motilities in isolated rat hepatocytes." J. Clin. Electron Microscopy. 22. 738-739 (1989)

Watanabe, N.、Miyairi, M.、Suyama, K.、Oshio, C.、Oda, M. 和 Tsuchiya, M.：“牛磺石胆酸盐对离体大鼠肝细胞胆小管运动的影响。”

Masaya,Oda: Journal of Clinical Electron Microscopy.

Masaya，Oda：临床电子显微镜杂志。

織田 正也: "胆汁分泌とその異常" 臨床消化器内科. 2. 140 (1987)

Masaya Oda：“胆汁分泌及其异常”《临床胃肠病学》2. 140 (1987)。

Masaya Oda: "Electron microscopic changes of intrahepatocellular calcium ion distribution in bile canalicular contractionsーdigital image analysis combined with furaー2 fluorescenceー." Hepatology. 10. 635 (1989)

Masaya Oda：“胆小管收缩时肝细胞内钙离子分布的电子显微镜变化－数字图像分析结合fura-2荧光－”，《肝病学》10. 635（1989）。