Quantitative assessment of canalicular bile formation in isolated hepatocyte couplets using microscopic optical planimetry.
Quantitative assessment of canalicular bile formation in isolated hepatocyte couplets using microscopic optical planimetry.
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使用显微光学面积测量法定量评估分离的肝细胞对中胆管胆汁的形成。
DOI：
10.1172/jci113919
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Boyer,JL
中科院分区：
文献类型：
--
作者：
Gautam,A;Ng,OC;Strazzabosco,M;Boyer,JL
分离的大鼠肝细胞偶联（IRHC）是胆汁分泌的主要单位，在培养中随着时间的推移在封闭的小管空间中积累液体。我们通过使用高分辨率Nomarski光学器件的视频显微镜光学测面术测量小管空间体积的变化，定量了培养4-8小时的IRHC中小管分泌的速率。电子显微镜形态学研究显示，在培养4-6小时后，小管膜面积显着增加。基础L-15培养基中的小管分泌（3.8 +/- 1.3 fl/min）随着利胆胆汁盐（10 μ M）、牛磺胆汁酸盐和熊去氧胆汁酸盐（各14 +/- 7 fl/min）而显着增加。暴露于胆汁酸后的分泌率与刺激前的小管表面积直接相关。相反，刺激后的扩张时间与初始泪小管体积成反比。与牛磺胆酸盐相比，熊去氧胆酸在10-，100-或200-microM浓度下未能产生高胆汁分泌，无论是在正常还是牛磺酸耗尽的IRHC中。目前的研究结果表明，在基础状态下和胆汁酸刺激后，可以通过连续光学面积测量法定量IRHC中的小管胆汁分泌率。此外，在该模型中，熊去氧胆酸盐不会在小管水平急性诱导胆汁分泌过多。相反，牛磺胆酸和熊去氧胆酸诱导分泌的小管膜的表面积成比例。IRHC是一个有用的模型来识别小管胆汁分泌和研究小管胆汁形成。图像
Isolated rat hepatocyte couplets (IRHC) are primary units of bile secretion that accumulate fluid in an enclosed canalicular space with time in culture. We have quantitated the rate of canalicular secretion in IRHC cultured for 4-8 h by measuring the change in canalicular space volume by video-microscopic optical planimetry using high resolution Nomarski optics. Electron microscopic morphometric studies revealed significant increases in canalicular membrane area after 4-6 h in culture. Canalicular secretion in basal L-15 medium (3.8 +/- 1.3 fl/min) increased significantly with the choleretic bile salts (10 microM), taurocholate, and ursodeoxycholate (14 +/- 7 fl/min each). Secretion rates after exposure to bile acids correlated directly with the canalicular surface area before stimulation. In contrast, expansion times after stimulation varied inversely with initial canalicular volumes. Ursodeoxycholic acid failed to produce a hypercholeresis at 10-, 100-, or 200-microM concentrations compared with taurocholate, either in normal or taurine-depleted IRHC. The present findings establish that rates of canalicular bile secretion can be quantitated in IRHC by serial optical planimetry, both in the basal state and after stimulation with bile acids. Furthermore, ursodeoxycholate does not acutely induce hypercholeresis at the canalicular level in this model. Rather, both taurocholic and ursodeoxycholic acids induced secretion in proportion to the surface area of the canalicular membrane. The IRHC are a useful model to identify canalicular choleretics and for studies of canalicular bile formation.Images