Clark D, Brinkman M, Neville S D, Grivell T, Phillips J
CSIRO (Australia), Division of Human Nutrition, Adelaide.
Biochim Biophys Acta. 1992 Jan 16;1098(2):240-6. doi: 10.1016/s0005-2728(05)80342-1.
When hepatocytes prepared from 24-h-fasted rats were washed, suspended and incubated in Krebs-Henseleit bicarbonate-buffered saline, the endogenous rates of O2 consumption and heat production were 2.13 +/- 0.13 mumol/min per g wet wt. and 1.00 +/- 0.05 J/min per g wet wt. respectively. The inclusion of 2.5% (w/v) defatted and dialysed bovine serum albumin in either the cell suspension (washing) buffer or the cell incubation buffer produced a 20-25% increase in O2 consumption and heat production: these rates were increased by an additional 20-25% when the albumin (2.5%) was present in both the cell suspension and the cell incubation buffers. There was an inverse relationship between the increases in O2 consumption and heat production and the leakage of lactate dehydrogenase from the isolated hepatocytes: the inclusion of purified bovine serum albumin decreased lactate dehydrogenase leakage from 40% to 15% of total enzyme content. The calorimetric-respirometric ratios for hepatocytes incubated both in the absence (-461 +/- 19 kJ/mol O2) and presence (-477 +/- 8 kJ/mol O2) of the purified protein are very similar to the theoretical, thermochemically derived oxycaloric equivalents.
将禁食24小时的大鼠制备的肝细胞洗涤、悬浮并在Krebs-Henseleit碳酸氢盐缓冲盐溶液中孵育时，氧气消耗和产热的内源性速率分别为每克湿重2.13±0.13 μmol/分钟和每克湿重1.00±0.05 J/分钟。在细胞悬浮（洗涤）缓冲液或细胞孵育缓冲液中加入2.5%（w/v）脱脂并透析的牛血清白蛋白，会使氧气消耗和产热增加20%-25%：当白蛋白（2.5%）同时存在于细胞悬浮液和细胞孵育缓冲液中时，这些速率会再增加20%-25%。氧气消耗和产热的增加与分离的肝细胞中乳酸脱氢酶的泄漏呈负相关：加入纯化的牛血清白蛋白可使乳酸脱氢酶泄漏从总酶含量的40%降至15%。在不存在（-461±19 kJ/mol O2）和存在（-477±8 kJ/mol O2）纯化蛋白的情况下孵育的肝细胞的量热-呼吸比与理论上热化学推导的氧热当量非常相似。
