McKay N D, Robinson B, Brodie R, Rooke-Allen N
Biochim Biophys Acta. 1983 Apr 5;762(2):198-204. doi: 10.1016/0167-4889(83)90071-x.
Human skin fibroblast cultures, seeded at 10(5) cells/5 cm plate and allowed to grow to confluence at approx. 10(6) cells/5 cm plate, utilized a glycolytic mode of metabolism where the ratio of glucose utilized to lactate produced wa 0.62 +/- 0.05 (Zielke, R.H., Ozand, P.T., Tyldon, J.I., Sevdalian, D.A. and Cornblath, M. (1976) Proc. Natl. Acad. Sci. U.S.A. 73, 4110-4114) (mean +/- S.E.). When the glucose in the medium was exhausted, the lactate produced during the highly glycolytic phase was then reutilized. In monolayer cultures that had been washed with phosphate-buffered saline, rates of glucose utilization were measured at 0.25 and 2 mM glucose by monitoring the appearance of 3H2O from [5-3H]glucose. Rate of utilization for each concentration of glucose decreased markedly as the cultures became more confluent. This decrease also correlated with a reduced ability to transport glucose as measured by 2-deoxy-[3H]glucose uptake in washed monolayer cultures. In washed confluent culture of fibroblasts, glucose utilization was markedly decreased by the presence of pyruvate and lactate but not by glutamine. The respiratory inhibitors, rotenone and antimycin, did not increase the rate of glucose utilization except when added in combination with pyruvate. We conclude that cultured skin fibroblasts possess a highly glycolytic mode of metabolism but that this mode can become more oxidative in the presence of sufficient quantities of pyruvate and lactate.
人皮肤成纤维细胞培养物以10⁵个细胞/5厘米培养皿接种,在约10⁶个细胞/5厘米培养皿时生长至汇合,采用糖酵解代谢模式,其中消耗的葡萄糖与产生的乳酸的比率为0.62±0.05(齐尔克,R.H.,奥赞德,P.T.,泰勒登,J.I.,塞夫达利安,D.A.和科恩布拉思,M.(1976年)美国国家科学院院刊73,4110 - 4114)(平均值±标准误)。当培养基中的葡萄糖耗尽时,在高度糖酵解阶段产生的乳酸随后被重新利用。在用磷酸盐缓冲盐水洗涤过的单层培养物中,通过监测[5 - ³H]葡萄糖中³H₂O的出现来测量在0.25和2 mM葡萄糖时的葡萄糖消耗率。随着培养物变得更加汇合,每种葡萄糖浓度的消耗率显著下降。这种下降也与通过在洗涤过的单层培养物中摄取2 - 脱氧 - [³H]葡萄糖测量的葡萄糖转运能力降低相关。在洗涤过的成纤维细胞汇合培养物中,丙酮酸和乳酸的存在显著降低了葡萄糖消耗,但谷氨酰胺没有。呼吸抑制剂鱼藤酮和抗霉素,除非与丙酮酸联合添加,否则不会增加葡萄糖消耗率。我们得出结论,培养的皮肤成纤维细胞具有高度糖酵解的代谢模式,但在存在足够量的丙酮酸和乳酸时,这种模式可以变得更具氧化性。