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豚鼠肝脏实质细胞的分离制备及特性研究

Preparation and characterization of isolated parenchymal cells from guinea pig liver.

作者信息

Elliott K R, Pogson C I

出版信息

Mol Cell Biochem. 1977 May 31;16(1):23-9. doi: 10.1007/BF01769835.

Abstract
  1. A method is described for the preparation of isolated cells from guinea pig liver. This involved perfusion in situ, in the non-physiological direction, with collagenase. 2. The cell yield was 20--30%, comparable with those from the livers of other species. 3. The ratio of lactate dehydrogenase to glutamate dehydrogenase in the cells was similar to that in vivo, indicating that there was negligible leakage of cytoplasmic enzymes. 4. The concentrations of K+ and adenine nucleotides were initially lower than in the perfused liver; normal values were obtained on incubation, particularly in the presence of substrate. 5. The L-lactate: pyruvate ratio is 16:1, close to established values. The total beta-hydroxybutyrate: acetoacetate ratio indicates that the mitochondrial redox state is more oxidised than in the perfused liver, but the intracellular ratio is similar to that of the intact liver. 6. Rates of gluconeogenesis and ureogenesis, are within the physiological range. Maximal gluconeogeneis from L-lactate was preceded by a lag period. L-lysine stimulated glucose production from L-lactate but did not abolish the lag phase. 7. The effects of aminooxyacetate and octanoate on L-lactate gluconeogenesis were similar to those in the perfused liver.
摘要
  1. 描述了一种从豚鼠肝脏制备分离细胞的方法。这包括用胶原酶在非生理方向原位灌注。2. 细胞产量为20% - 30%,与其他物种肝脏的产量相当。3. 细胞中乳酸脱氢酶与谷氨酸脱氢酶的比率与体内相似,表明细胞质酶的泄漏可忽略不计。4. 钾离子和腺嘌呤核苷酸的浓度最初低于灌注肝脏中的浓度;孵育后可达到正常值,尤其是在有底物存在的情况下。5. L-乳酸:丙酮酸的比率为16:1,接近既定值。总β-羟基丁酸:乙酰乙酸的比率表明线粒体氧化还原状态比灌注肝脏中更氧化,但细胞内比率与完整肝脏相似。6. 糖异生和尿素生成速率在生理范围内。从L-乳酸进行最大糖异生前有一个滞后期。L-赖氨酸刺激L-乳酸生成葡萄糖,但并未消除滞后期。7. 氨基氧乙酸和辛酸对L-乳酸糖异生的影响与灌注肝脏中的相似。

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