Tanyalçin T, Deveci R, Kutay F, Karaçali S, Sözmen E Y
Ege University School of Medicine, Department of Biochemistry, Bornova, Izmir-Turkey.
Biochem Mol Biol Int. 1995 Mar;35(3):517-27.
We described a procedure for the preservation of rat liver which makes possible the isolation of plasma membranes after 10 days storage at -70 degrees C. The yield of plasma membranes obtained from the liver tissue kept at -70 degrees C for 10 days (3.43 +/- 0.08 mg protein/10 g wet liver) was not different statistically (P > 0.05) from the yield of freshly obtained plasma membranes (3.32 +/- 0.05 mg protein/10 g wet liver). However, a significantly low yield (2.65 +/- 0.08; P < 0.01) was obtained from 90 days stored rat liver when compared with the immediate isolation. Plasma membrane Na+, K+ ATPase and 5'nucleotidase activities of the stored liver for 10 days were not different statistically (P > 0.05) from the enzyme activities of the freshly isolated membrane fractions. In contrast there was a significant decrease (p < 0.0001) in the activities of both plasma membrane Na+, K+ ATPase and 5'nucleotidase activities of 90 days stored rat liver at -70 degrees C when compared with immediate isolation. Considering the electron microscopic findings; we observed that the preservation of the integrity of the plasma membrane fractions obtained from fresh and frozen livers for 10 and 90 days seemed to be parallel to the biochemical results. Therefore we suggest that, storage of rat liver tissue for 10 days make feasible to maintain the experimental design and give convenience for obtaining intact plasma membrane fractions.
我们描述了一种大鼠肝脏保存方法，该方法能使肝脏在-70℃储存10天后分离得到质膜。从在-70℃保存10天的肝脏组织中获得的质膜产量（3.43±0.08mg蛋白质/10g湿肝）与新鲜获得的质膜产量（3.32±0.05mg蛋白质/10g湿肝）在统计学上无差异（P>0.05）。然而，与立即分离相比，90天储存的大鼠肝脏获得的产量显著较低（2.65±0.08；P<0.01）。储存10天的肝脏质膜Na⁺、K⁺-ATP酶和5'-核苷酸酶活性与新鲜分离的膜组分的酶活性在统计学上无差异（P>0.05）。相比之下，与立即分离相比，在-70℃储存90天的大鼠肝脏质膜Na⁺、K⁺-ATP酶和5'-核苷酸酶活性均显著降低（P<0.0001）。考虑到电子显微镜检查结果；我们观察到，从新鲜和冷冻肝脏中获得的质膜组分在储存10天和90天后其完整性的保持似乎与生化结果一致。因此我们建议，大鼠肝脏组织储存10天能够维持实验设计，并便于获得完整的质膜组分。
