Kohmoto M, Tanaka J, Yoshida M, Kasamatsu T, Fujita K I, Zhong W B, Kaidou T, Tamura J, Imamura M
First Department of Surgery, Faculty of Medicine, Kyoto University, Japan.
Eur Surg Res. 1994;26(4):221-9. doi: 10.1159/000129339.
Dynamic changes in liver plasma membrane fluidity caused by regeneration and atrophy were assessed in rats following portal branch ligation (PBL). The portal branch, which perfuses 70% of the liver, was ligated with 5-0 prolene, and liver plasma membranes were isolated by ultracentrifugation. The membrane fluorescence polarization was measured as an index of membrane fluidity using 1,6-diphenyl-1,3,5-hexatriene (DPH) as the probe dye. In nonligated lobes, a significant decrease in fluorescence polarization was observed 12 and 24 h after PBL (0.171 +/- 0.004, p < 0.01 and 0.165 +/- 0.005, p < 0.001, respectively) as compared to the controls (0.181 +/- 0.002). The fluorescence polarization values then gradually returned to near control levels. In contrast, in the ligated lobes, the fluorescence polarization had increased by 12 hours after PBL (0.196 +/- 0.002, p < 0.01), and remained significantly elevated (p < 0.01) for up to 1 week after PBL, gradually returning to control levels within 3 weeks. The membrane composition was also evaluated by analyzing the cholesterol/phospholipid (C/P) ratio. A significant increase in the C/P ratio was detected in the ligated lobes 12 h and 3 days after PBL, but there was no significant difference in fluorescence polarization values between nonligated lobes and controls. These results suggest that alterations in membrane fluidity play an important role in the regenerative and atrophic processes of the liver following portal branch ligation.
在大鼠门静脉分支结扎(PBL)后,评估再生和萎缩引起的肝细胞膜流动性的动态变化。用5-0聚丙烯缝线结扎灌注70%肝脏的门静脉分支,通过超速离心分离肝细胞膜。使用1,6-二苯基-1,3,5-己三烯(DPH)作为探针染料,测量膜荧光偏振作为膜流动性指标。在未结扎叶中,PBL后12小时和24小时观察到荧光偏振显著降低(分别为0.171±0.004,p<0.01和0.165±0.005,p<0.001),与对照组(0.181±0.002)相比。然后荧光偏振值逐渐恢复到接近对照水平。相反,在结扎叶中,PBL后12小时荧光偏振增加(0.196±0.002,p<0.01),并且在PBL后长达1周保持显著升高(p<0.01),在3周内逐渐恢复到对照水平。还通过分析胆固醇/磷脂(C/P)比率评估膜组成。在PBL后12小时和3天,在结扎叶中检测到C/P比率显著增加,但在未结扎叶和对照组之间的荧光偏振值没有显著差异。这些结果表明,膜流动性的改变在门静脉分支结扎后肝脏的再生和萎缩过程中起重要作用。