Masuno H, Tsujita T, Nakanishi H, Yoshida A, Fukunishi R, Okuda H
J Lipid Res. 1984 May;25(5):419-27.
The triglyceride lipase (TGL) activity of liver homogenates of mice with Sarcoma 180 was measured. The liver homogenate of normal or tumor-bearing mice was treated with 0.25% Triton X-100 and centrifuged at 100,000 g for 60 min, and the supernatant was applied to a heparin-Sepharose column. In normal mice, most of the TGL activities in the supernatant was eluted with 0.75 M NaCl from the column. In mice with Sarcoma 180, the TGL gave two peaks on heparin-Sepharose column chromatography, which were eluted with 0.75 M and 1.5 M NaCl, respectively. The activity in the first peak (0.75 M NaCl eluate) decreased; that in the second peak (1.5 M NaCl eluate) increased, and the ratio of the second peak to the first peak increased during tumor development. The livers of normal mice and mice on day 10 after tumor inoculation were perfused with heparin. The highest rate of the TGL release occurred within 1 min of heparin perfusion, and the bulk of heparin-releasable activity appeared within 2 min of perfusion in both normal and tumor-bearing mice. The TGL activity in liver perfusate of tumor-bearing mice, as well as that of liver homogenate, was resolved on a heparin-Sepharose column into two peaks, which were eluted with 0.75 M and 1.5 M NaCl, and most of the activity was eluted with 1.5 M NaCl. The nature of the TGL activity eluted from a heparin-Sepharose column was investigated. In both liver homogenates and liver perfusates, the first peak did not require serum for maximal activity and was relatively resistant to a high concentration of NaCl or protamine sulfate.(ABSTRACT TRUNCATED AT 250 WORDS)
对患有肉瘤180的小鼠肝脏匀浆中的甘油三酯脂肪酶(TGL)活性进行了测定。将正常或荷瘤小鼠的肝脏匀浆用0.25% Triton X - 100处理,并在100,000 g下离心60分钟,然后将上清液应用于肝素 - 琼脂糖柱。在正常小鼠中,上清液中的大部分TGL活性用0.75 M NaCl从柱上洗脱下来。在患有肉瘤180的小鼠中,TGL在肝素 - 琼脂糖柱色谱上出现两个峰,分别用0.75 M和1.5 M NaCl洗脱。第一个峰(0.75 M NaCl洗脱液)中的活性降低;第二个峰(1.5 M NaCl洗脱液)中的活性增加,并且在肿瘤发展过程中第二个峰与第一个峰的比例增加。对正常小鼠和接种肿瘤后第10天的小鼠肝脏进行肝素灌注。TGL释放的最高速率在肝素灌注后1分钟内出现,并且在正常和荷瘤小鼠中,大部分可被肝素释放的活性在灌注后2分钟内出现。荷瘤小鼠肝脏灌注液中的TGL活性以及肝脏匀浆中的TGL活性在肝素 - 琼脂糖柱上均被分离为两个峰,分别用0.75 M和1.5 M NaCl洗脱,并且大部分活性用1.5 M NaCl洗脱。对从肝素 - 琼脂糖柱洗脱的TGL活性的性质进行了研究。在肝脏匀浆和肝脏灌注液中,第一个峰的最大活性不需要血清,并且对高浓度的NaCl或硫酸鱼精蛋白相对抗性较强。(摘要截短至250字)