Adult rat hepatocytes in primary culture. VI. Developmental changes in alcohol dehydrogenase activity and ethanol conversion during the growth cycle.
Adult rat hepatocytes in primary culture. VI. Developmental changes in alcohol dehydrogenase activity and ethanol conversion during the growth cycle.
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原代培养的成年大鼠肝细胞。
DOI：
10.1111/j.1530-0277.1982.tb05382.x
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Leffert,HL
中科院分区：
文献类型：
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作者：
Lad,PJ;Shier,WT;Skelly,H;DeHemptinne,B;Leffert,HL
具有增殖能力的成年大鼠肝细胞原代“单层”培养物显示出一系列分化功能，其特征在于乙醇脱氢酶活性的生长状态依赖性表达（ADH；EC 1.1.1.1）。肝细胞分离后和体外滞后期早期，“每个细胞”的 ADH 相当于对照成人肝组织的水平。进入对数增长并进行时，ADH 双相下降至对照水平的约 2%（半衰率，R[消失]= 3.4 和 7.4 小时；第 0 天（分别为半衰率，R[消失]= 3.4 和 7.4）。随着肝细胞进入并保持静止期，ADH 双相增加（R[消失]= 3.9 和 11.1）最终，当测量 [14C] 乙醇转化为 CHCl3:CH30H (2:1) 可提取物质的速率时，最终，10 至 13 天的肝细胞培养物转化乙醇或 [14C] 乙酸盐的速度比稳定期 3T3 成纤维细胞快 10 倍。 （分别为 3000 或 12,000 cpm/106 细胞/24 小时与 200 或 800 cpm/106 细胞/24 小时）。与非肝细胞系统不同，成熟肝细胞培养物中的两种转化过程均取决于细胞外葡萄糖水平。此外，根据生长状态，肝细胞培养物中的 [14C] 乙醇转化可被 1 mil 吡唑（一种特定的 ADH）抑制高达 73%。在较高的初始乙醇水平（5000 万）下，吡唑阻断无效。结果表明，在这种长期培养系统中，正常成年肝细胞保留了非 ADH 乙醇转化酶和药理学敏感的 ADH，培养物中后一种酶的活性水平的变化似乎模拟了完整动物中肝脏 ADH 的正常发育变化。
Proliferation‐competent primary ‘monolayer’ cultures of adult rat hepatocytes that display a repertoire of differentiated functions have been characterized for growth state‐dependent expression of alcohol dehydrogenase activity (ADH; EC 1.1.1.1). Following hepa‐tocyte isolation and early during in vitro lag phase, ADH ‘per cell’ is equivalent to control adult liver tissue levels. Upon entry into and progression of logarithmic growth, ADH declines biphasically to about 2% of control levels (halftime rate, R[disappearance]= 3.4 and 7.4 hr; days 0‐is(halftime rate, R[disappearance]= 3.4and 7.4, respectively). increases biphasically as hepatocytes enter and remain in stationary phase (R[disappear]= 3.9 and 11.1 hr; days 6–10 and 10–14, respectively). Ultimately, control ADH is restored. Similar ‘U‐shaped’ curves occur when rates of conversion of [14C]ethanol into CHCI3:CH30H (2:1) extractable material are measured. By this criterion, 10‐ to 13‐day‐old hepatocyte cultures convert ethanol or [14C]acetate 10 times faster than stationary‐phase 3T3 fibroblasts' (3000 or 12,000 cpm/106cells/24 hours versus 200 or 800 cpm/106cells/24 hr, respectively). Both conversion processes in matured hepatocyte cultures depend on extracellular glucose levels unlike the nonhepatocyte system. Furthermore, depending on the growth state, [14C]ethanol conversion in hepatocyte cultures is inhibited up to 73% by 1 mil pyrazole, a specific ADH inhibitor. At higher initial ethanol levels (50 mil), pyrazole blockade is ineffective. The results indicate that, in this long‐term culture system, normal adult hepatocytes retain non‐ADH ethanol‐converting enzymes and pharmacologically‐sensitive ADH. Changes in the latter enzyme's activity levels in culture appear to simulate normal developmental changes in hepatic ADH in the intact animal.