Boot J H
Department of Veterinary Pharmacology, Pharmacy & Toxicology, University Utrecht, The Netherlands.
Cell Struct Funct. 1995 Jun;20(3):233-8. doi: 10.1247/csf.20.233.
To investigate the importance of SH-groups in the energy metabolism of liver cells, isolated rat hepatocytes were exposed to various SH-blocking compounds. After 1.0 hr exposure, the cells were analyzed for the content of glycogen, lactate, pyruvate, ATP and the rate of oxygen consumption. Without affecting the cell viability, PCMB, PCMBS, mersalyl, NEM, DTP and DSF were found to decrease glycogen levels, whereas the disulphide reagent CDPS did not affect this endogenous energy reserve. Lactate and pyruvate levels were decreased by the organic mercury compounds, whereas NEM, DTP and DSF stimulated the formation of lactate, without affecting the levels of pyruvate. In both situations the oxygen consumption was slightly decreased. The FCCP uncoupled oxygen consumption was not affected. Up to the point of loss of cell viability, as measured by trypan blue exclusion and LDH-leakage, the liver cells maintained their ATP levels, during exposure to the various SH-reagents. In conclusion, the results with organic mercury compounds suggest a reaction of these agents with SH-groups in the outer membrane of cells, having an inhibiting effect on the glucose uptake. The most prominent effect of DTP, DSF and NEM was an increased lactate formation, implying an intracellular effect, most probably in the TCA-cycle.
为了研究巯基在肝细胞能量代谢中的重要性，将分离的大鼠肝细胞暴露于各种巯基阻断化合物。暴露1.0小时后，分析细胞中的糖原、乳酸、丙酮酸、ATP含量以及耗氧率。在不影响细胞活力的情况下，发现对氯汞苯甲酸（PCMB）、对氯汞苯磺酸钠（PCMBS）、汞撒利、N-乙基马来酰亚胺（NEM）、二巯基丙醇（DTP）和二硫苏糖醇（DSF）会降低糖原水平，而二硫化物试剂CDPS不影响这种内源性能量储备。有机汞化合物会降低乳酸和丙酮酸水平，而NEM、DTP和DSF会刺激乳酸的形成，而不影响丙酮酸水平。在这两种情况下，耗氧量均略有下降。FCCP解偶联的耗氧量不受影响。通过台盼蓝排斥法和乳酸脱氢酶泄漏测定，直至细胞活力丧失点，肝细胞在暴露于各种巯基试剂期间维持其ATP水平。总之，有机汞化合物的结果表明这些试剂与细胞外膜中的巯基发生反应，对葡萄糖摄取具有抑制作用。DTP、DSF和NEM最显著的作用是乳酸形成增加，这意味着细胞内效应，很可能发生在三羧酸循环中。
