31P-NMR spectroscopy of perifused rat hepatocytes immobilized in agarose threads: application to chemical-induced hepatotoxicity.
31P-NMR spectroscopy of perifused rat hepatocytes immobilized in agarose threads: application to chemical-induced hepatotoxicity.
复制标题
固定在琼脂糖线中的灌注大鼠肝细胞的 31P-NMR 光谱：应用于化学诱导的肝毒性。
DOI：
10.1016/0925-4439(92)90089-6
复制
发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Ho,C
中科院分区：
文献类型：
--
作者：
Farghali,H;Rossaro,L;Gavaler,JS;VanThiel,DH;Dowd,SR;Williams,DS;Ho,C
已发现由琼脂糖线固定的分离大鼠肝细胞组成的系统连续灌注含氧Krebs-Henseleit（KH）溶液可保持细胞活力，并具有良好的代谢活性超过6 h。通过在4.7特斯拉下的磷-31核磁共振（31 P-NMR）光谱、通过测量氧消耗以及通过乳酸脱氢酶（LD）和丙氨酸氨基转移酶（ALT）的泄漏来监测肝细胞。所获得的数据与在体外分离的灌注全肝中发现的那些数据相当。考察了烯丙醇（AA）、乙醇和对乙酰氨基苯酚（AP）的影响。225 μM AA溶液灌流90 min后，~（31）P-NMR谱中三磷酸腺苷（ATP）的β-磷酸共振消失，LD泄漏增加7倍，耗氧量减少70%。乙醇（1.0M）灌流90 min使β-ATP信号强度比降低20%，磷酸单酯（PME）信号降低50%，无机磷酸盐（Pi）信号降低33%（P< 0.05）。AP（10 mM）仅引起轻度肝细胞损伤。结果表明，灌流固定化肝细胞可以用作肝脏模型，以评估各种化学品和其他外源性物质的影响，通过NMR光谱。
A system consisting of isolated rat hepatocytes immobilized in agarose threads continuously perifused with oxygenated Krebs-Henseleit (KH) solution has been found to maintain cell viability with excellent metabolic activity for more than 6 h. The hepatocytes were monitored by phosphorus-31 nuclear magnetic resonance (31P-NMR) spectroscopy at 4.7 Tesla, by measurement of oxygen consumption and by the leakage of lactate dehydrogenase (LD) and alanine aminotransferase (ALT). The data obtained were comparable to those found for an isolated perfused whole liver in vitro. The effects of allyl alcohol (AA), ethanol, and 4-acetaminophenol (AP) were examined. A solution of 225 μM AA perifused for 90 min caused the disappearance of the β-phosphate resonance of adenosine triphosphate (ATP) in the31P-NMR spectra, a 7-fold increase in LD leakage and a 70% reduction in oxygen consumption. Ethanol (1.0 M) perifused for 90 min reduced the β-ATP signal intensity ratio by 20%, the phosphomonoester (PME) signal by 50% and inorganic phosphate (Pi) by 33% (P< 0.05). AP (10 mM) caused only mild liver-cell damage. The results demonstrate that perifused immobilized hepatocytes can be used as a liver model to assess the effects of a wide range of chemicals and other xenobiotics by NMR spectroscopy.