Bibby Susan R S, Jones Deborah A, Ripley Ruth M, Urban Jill P G
University Laboratory of Physiology, Oxford University, Oxford, United Kingdom.
Spine (Phila Pa 1976). 2005 Mar 1;30(5):487-96. doi: 10.1097/01.brs.0000154619.38122.47.
In vitro measurements of metabolic rates of isolated bovine nucleus pulposus cells at varying levels of oxygen, glucose, and pH.
To obtain quantitative information on the interactions between oxygen and glucose concentrations and pH, and the rates of oxygen and glucose consumption and lactic acid production, for disc nucleus cells.
Disc cells depend on diffusion from blood vessels at the disc margins for supply of nutrients. Loss of supply is thought to lead to disc degeneration, but how loss of supply affects nutrient concentrations in the disc is not known; nutrient concentrations within discs can normally only be calculated, because concentration measurements are invasive. However, realistic predictions cannot be made until there are data from measurements of metabolic rates at conditions found in the disc in vivo, i.e., at low levels of oxygen, glucose, and pH.
A metabolism chamber was designed to allow simultaneous recording of oxygen and glucose concentrations and of pH. These concentrations were measured electrochemically with custom-built glucose and oxygen sensors; lactic acid was measured biochemically. Bovine nucleus pulposus cells were isolated and inserted into the chamber, and simultaneous rates of oxygen and glucose consumption and of lactic acid production were measured over a range of glucose, oxygen, and pH levels.
There were strong interactions between rates of metabolism and oxygen consumption and pH. At atmospheric oxygen levels, oxygen consumption rate at pH 6.2 was 32% of that at pH 7.4. The rate fell by 60% as oxygen concentration was decreased from 21 to 5% at pH 7.4, but only by 20% at pH 6.2. Similar interactions were seen for lactic acid production and glucose consumption rates; we found that glycolysis rates fell at low oxygen and glucose concentrations and low pH. Equations were derived that satisfactorily predict the effect of nutrient and metabolite concentrations on rates of lactic acid production rate and oxygen consumption.
Disc cell metabolism in air and at pH 7.4 differs markedly from that found in the disc nucleus in vivo, where low levels of oxygen, glucose, and pH all coexist.
在不同氧气、葡萄糖和pH水平下对分离的牛髓核细胞代谢率进行体外测量。
获取关于椎间盘髓核细胞中氧气与葡萄糖浓度及pH之间的相互作用,以及氧气和葡萄糖消耗率与乳酸生成率的定量信息。
椎间盘细胞依赖于从椎间盘边缘血管的扩散来获取营养物质供应。营养物质供应的丧失被认为会导致椎间盘退变,但供应丧失如何影响椎间盘中的营养物质浓度尚不清楚;由于浓度测量具有侵入性,椎间盘中的营养物质浓度通常只能通过计算得出。然而,在获得体内椎间盘实际存在的条件下(即低氧、低糖和低pH水平)的代谢率测量数据之前,无法做出实际的预测。
设计了一个代谢室,用于同时记录氧气和葡萄糖浓度以及pH值。这些浓度通过定制的葡萄糖和氧气传感器进行电化学测量;乳酸通过生化方法测量。分离牛髓核细胞并将其放入代谢室,在一系列葡萄糖、氧气和pH水平下,同时测量氧气和葡萄糖消耗率以及乳酸生成率。
代谢率与氧气消耗率及pH之间存在强烈的相互作用。在大气氧水平下,pH 6.2时的氧气消耗率是pH 7.4时的32%。在pH 7.4时,当氧气浓度从21%降至5%时,氧气消耗率下降了60%,但在pH 6.2时仅下降了20%。乳酸生成率和葡萄糖消耗率也存在类似的相互作用;我们发现糖酵解率在低氧、低糖浓度和低pH时下降。推导得出的方程能够令人满意地预测营养物质和代谢物浓度对乳酸生成率和氧气消耗率的影响。
在空气中且pH为7.4时的椎间盘细胞代谢与体内椎间盘髓核中的代谢明显不同,体内椎间盘髓核中低氧、低糖和低pH同时存在。