Cationic colloidal silica membrane perturbation as a means of examining changes at the sinusoidal surface during liver regeneration
Cationic colloidal silica membrane perturbation as a means of examining changes at the sinusoidal surface during liver regeneration
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DOI：
10.1016/s0002-9440(10)65464-8
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发表时间：
1999-11-01
影响因子：
6
通讯作者：
Enomoto, K
中科院分区：
文献类型：
--
作者：
Stolz, DB;Ross, MA;Enomoto, K
通过采用阳离子胶体二氧化硅膜密度扰动技术，我们研究了生长因子受体和细胞外基质（ECM）的变化在肝窦表面在大鼠肝再生72小时后70%部分肝切除术（PHx）。在PHx后的这个时候，肝细胞分裂大部分已经消退，白色窦状内皮细胞（SEC）增殖开始，导致无血管肝细胞岛。由于肝SEC表面的不连续性质，当用胶体灌注肝时，SEC下面的ECM蛋白，如SEC管腔膜蛋白，可用于吸附到带电的二氧化硅珠。随后的肝匀浆和密度离心产生两个单独的级分，富集SEC以及肝细胞基底外侧膜特异性蛋白质，其高达全肝裂解物的50倍。这种技术有助于检查蛋白质组成的变化，这些变化影响或发生在肝脏再生过程中SEC有丝分裂和迁移的结果。当ECM和受体蛋白SEC富集的馏分进行了检查，通过Western免疫印迹，尿激酶纤溶酶原激活物受体，纤连蛋白，纤溶酶增加SEC表面PHx后72小时。与对照肝脏相比，PHx后72小时，表皮生长因子受体、纤溶酶原、骨钙素（分泌蛋白，酸性且富含半胱氨酸，也称为骨粘连蛋白或BM 40）和胶原IV减少，纤维蛋白原亚基和c-Met表达保持恒定。这些结果显示了阳离子胶态二氧化硅膜分离方案的有用性。它们还显示出对表面成分的相当大的调节，这些成分可能在肝窦重建期间的肝脏再生末期调节血管生成过程。
By employing the cationic colloidal silica membrane density perturbation technique, we examined growth factor receptor and extracellular matrix (ECM) changes at the sinusoidal surface during rat liver regeneration 72 hours after 70% partial hepatectomy (PHx). At this time after PHx, hepatocyte division has mostly subsided, white sinusoidal endothelial cell (SEC) proliferation is initiating, resulting in avascular hepatocyte islands. Because of the discontinuous nature of the surface of liver SEC, ECM proteins underlying the SEC, as web as SEC luminal membrane proteins, are available to absorption to the charged silica beads when the liver is perfused with the colloid. Subsequent liver homogenization and density centrifugation yield two separate fractions, enriched in SECs as well as hepatocyte basolateral membrane-specific proteins up to 50-fold over whole Liver lysates. This technique facilitates examination of changes in protein composition that influence or occur as a result of SEC mitogenesis and migration during regeneration of the liver. When ECM and receptor proteins from SEC-enriched fractions were examined by Western immunoblotting, urokinase plasminogen activator receptor, fibronectin, and plasmin increased at the SEC surface 72 hours after PHx. Epidermal growth factor receptor, plasminogen, SPARC (secreted protein, acidic and rich in cysteine, also called osteonectin or BM40), and collagen IV decreased, and fibrinogen subunits and c-Met expression remained constant 72 hours after PHx when compared to control liver. These results display the usefulness of the cationic colloidal silica membrane isolation protocol. They also show considerable modulation of surface components that may regulate angiogenic processes at the end stage of Liver regeneration during the reformation of sinusoids.