摘要
目的 研究维拉帕米及Na+ K+通道阻滞剂四乙基胺、奎尼丁和阿米洛利对低温保存成人肝细胞能量代谢的影响和对细胞的保护作用。方法 用胶原酶消化法制备成人肝细胞 ,置于分别加入维拉帕米、四乙基胺 +奎尼丁 +阿米洛利、四乙基胺 +奎尼丁 +阿米洛利 +维拉帕米的保存液中保存 ,高效液相色谱仪检测各组肝细胞中ATP、ADP及AMP的含量。结果 在维拉帕米实验组 ,ATP水平在实验过程中始终高于低温保存对照组 ;前 4个小时ADP平稳升高 ,以后逐渐减低 ;AMP呈缓慢升高趋势 ,至 12h达峰值 ;总腺苷酸含量总体呈下降趋势 ,但平台期延长 ,下降缓慢 ;能量转换率的变化与总腺苷酸含量相似。在加用维拉帕米和Na+ K+通道阻滞剂的联合用药组 ,ATP缓慢降低到初期的 5 0 %以下 ;ADP初期缓慢上升 ,4h后逐渐下降 ;AMP呈缓慢上升趋势 ;总腺苷酸含量和能量转换率均较维拉帕米组高 ,提示细胞的活性保持良好。结论 在低温保存成人肝细胞时加入维拉帕米和Na+ K+通道阻滞剂可降低肝细胞的能量消耗 ,它们对细胞的保护具有协同作用。
Objectives The effect of varapamil and Na/K ion blockers TEA,quinidine and amiloride on energy metabolism of human hepatocyte were studied during cold storage. Method Prepared isolated human hepatocyte, ATP,ADP,AMP detected by High Performance Liquid Chromatography(HPLC). Results In varapamil group,ATP level is higher than that of low temperature group at all phases. ADP level increase slowly during first 4 hours followed by a decrease phase. AMP level increase to ( 2.82 ± 0.04 )μmol/g wet weight steadily. TAN show a whole decrease phase, but have a prolonged plateau and a gentle slope of decreasing. EC is the same as that of TAN. In varapamil and Na/K ion channel blockers groups, ATP decrease slowly,and reach to less than 50% of primary level as the preservation time prolonged. ADP increase at the first 4 hours and decrease later,reaching the lowest level at the 12th hour about ( 0.80 ± 0.07 )μmol/g wet weight.AMP has a increasing tendency slowly; TAN and EC level are both higher than that of varapamil group which indicating the cell activity is maintained well. Conclusions Cryopreservation and varapamil could economize energy consumption. Varapamil, TEA,quinidine and amiloride also could decrease energy consumption synergetically and have protective effect on human hepatocyte during cold storage.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2001年第5期266-268,共3页
Chinese Journal of Organ Transplantation
基金
国家自然科学基金资助项目 (3 94 0 0 13 8)
