Dawson A G
Biochem J. 1972 Nov;130(2):525-32. doi: 10.1042/bj1300525.
A method is described whereby short fragments of rat kidney tubule were obtained when kidney slices were gently dispersed by exposure to collagenase and hyaluronidase. When suspended in buffered saline the fragmented tubules respired actively over a period of several hours, the rate of oxygen consumption being proportional to the amount of cell protein. Oxygen uptake was stimulated by the addition of glucose, lactate, butyrate, alpha-oxoglutarate and other substrates and was decreased by the omission of Ca(2+) from the suspending medium. With alpha-oxoglutarate as the added substrate, dinitrophenol strongly stimulated oxygen uptake. Dinitrophenol had a less-marked stimulatory effect when glucose was the added substrate, and inhibited respiration in the absence of added substrate. Oligomycin inhibited respiration and this inhibition was partially reversed by dinitrophenol. Fragmented tubules synthesized glucose from lactate at a high rate but this capacity for gluconeogenesis was abolished by dinitrophenol and by physically damaging the cells.
本文描述了一种方法,通过将肾切片暴露于胶原酶和透明质酸酶中使其轻轻分散,从而获得大鼠肾小管的短片段。当悬浮于缓冲盐溶液中时,这些片段化的肾小管在数小时内能够活跃地呼吸,耗氧速率与细胞蛋白量成正比。添加葡萄糖、乳酸盐、丁酸盐、α-酮戊二酸和其他底物可刺激氧气摄取,而从悬浮培养基中省略Ca(2+)则会降低氧气摄取。以α-酮戊二酸作为添加底物时,二硝基苯酚强烈刺激氧气摄取。当添加葡萄糖作为底物时,二硝基苯酚的刺激作用不太明显,并且在没有添加底物的情况下会抑制呼吸。寡霉素抑制呼吸,而二硝基苯酚可部分逆转这种抑制作用。片段化的肾小管能够以高速率从乳酸盐合成葡萄糖,但这种糖异生能力会被二硝基苯酚和物理损伤细胞所消除。