离体肝脏保存-再灌注损伤的发生机制及丹参的保护作用的实验研究  

Experimental study on the mechanism of the preservation-reperfusion injury and the protective effect of salvia miltiorrhiza in the isolated liver

作　　者：梁德森[1] 周德良 田茂霖[1] 张铁民[1] 吕海涛[1] 

机构地区：[1]哈尔滨医科大学第一临床医学院普外科,黑龙江哈尔滨150001 [2]肇州县人民医院,黑龙江肇州163000

出　　处：《哈尔滨医科大学学报》2006年第3期196-200,共5页Journal of Harbin Medical University

基　　金：黑龙江省科委"十五"攻关资助项目(GB02C142-01)

摘　　要：目的探讨羟自由基(.OH)与细胞凋亡、Bcl-2蛋白表达在大鼠肝脏不同保存时相再灌注过程中的变化规律及相互关系,阐明丹参对肝脏保存再灌注损伤的影响。方法以大鼠肝脏非循环灌注为模型,用高效液相色谱(HPLC)紫外检测法测定.OH生成物二羟苯甲酸(DHBA)含量,用原位末端标记法及电子显微镜观察细胞凋亡,用免疫组织化学法检测Bcl-2蛋白表达,观察了正常组(未保存)、对照组(乳酸林格液即LR液)及丹参组(LR液+丹参)保存鼠肝不同时相的效果。结果随着保存时间延长(18h内),.OH水平及细胞凋亡指数(AI)呈进行性升高,且两者呈显著正相关(r=0.81),相同保存时间丹参组明显低于对照组(P<0.05);而Bcl-2蛋白表达指数(BI)呈进行性降低,BI与.OH水平及AI呈显著负相关(r=-0.84),相同保存时间丹参组BI显著高于对照组(P<0.05)。丹参组ATP含量及分泌胆汁量明显高于对照组,丹参组流出液天门冬氨酸转氨酶(AST)明显低于对照组,差异有显著性(P<0.05)。结论.OH及细胞凋亡同时参与肝脏保存-再灌注损伤,丹参对该损伤具有明显的保护作用,其作用机制与抗氧自由基、抑制细胞凋亡、提高Bcl-2蛋白表达及改善低温保存肝脏的能量代谢作用有关。Objective To explore the dynamic changes of hydroxyl radical （· OH） and apoptosis in a rat liver preservation-reperfusion model, and to discuss the interrelations and the protective effects of salvia miltiorrhiza. Methods We researched the preservation effect of control group （with lactate Ringer＇ s solution）, DS group （with lactate Ringer＇ s solution added with salvia miltiorrhiza） and normal group （without preservation） by measuring the content of dihydroxybenzoic acid （DHBA） which was formed after reaction of ·OH with salicylic acid, apoptosis index （AI） and the expression index of Bcl-2 protein （BI） on isolated nonrecireulated perfusion rat liver model. Results · OH levels expressed as DHBAs, and AI enhanced markedly as the time of preservation prolonged. There was a significantly positive correlation between DHBA contents and AI. Both parameters in DS group were significantly lower than those in control group with the same preservation time （ P 〈 0.05）. The dynamic changes of BI were just on the contrary compared with both parameters. After the rat livers were preserved for 12h in LR solution with the addition of salvia miltionrrhiza, the contents of ATP, TAN, AEC and bile production were significantly higher than in the control group （ P 〈 0.05）. It was ＇also found that salvia miltionrrhiza had a marked protection against hydroxyl free radical and alleviated the injury on liver cells. Conclusion Both · OH and apoptosis participate liver preservation-reperfusion injury, and salvia miltiorrhiza may play an important part in alleviating the injury by a mechanism which correlates to the effect of resisting free radical, inhibiting the occurrence of apoptosis, enhancing the expression of Bcl-2 protein and improving the energy metabolism of cryopreservation liver.

关 键 词：自由基 细胞凋亡 BCL-2蛋白 能量代谢 丹参 

分 类 号：R-332[医药卫生] R285.5