磷酸肌酸对离体大鼠肝脏冷保存的保护作用

摘要

目的：探讨磷酸肌酸（CP）对大鼠离体肝脏冷保存的保护作用。方法建立大鼠肝脏单纯冷保存离体灌注模型，对照组予单纯威斯康星大学保存液（UW 液）灌注肝脏，低剂量组以UW 液为基液加入1 g／100 ml CP 灌注肝脏，中剂量组以 UW 液为基液加入2 g／100 ml CP 灌注肝脏；高剂量组以 UW 液为基液加入3 g／100 ml CP 灌注肝脏。各组大鼠肝脏分别于4℃相应灌注液中冷保存后0、6、12、18、24 h 共5个时间点，分别检测肝下下腔静脉内保存液的丙氨酸转氨酶（ALT）、乳酸脱氢酶（LDH）含量，检测肝脏组织丙二醛（MDA）含量、髓过氧化物酶（MPO）活性，观察肝脏组织肝细胞的凋亡指数（AI）和肝脏组织核因子-κB 阳性表达率，光学显微镜下观察肝脏组织的病理学变化。结果低、中、高剂量组大鼠肝脏在冷保存12 h 后，ALT 及 LDH 含量均低于对照组（均为 P ＜0.05）；冷保存18 h 后低、中、高剂量组大鼠肝脏组织的 MDA、MPO 含量均低于对照组（均为P ＜0.05）；在冷保存12 h 及18 h 时，低、中、高剂量组大鼠肝脏的肝细胞 AI 及核因子-κB 阳性表达率均低于对照组（均为 P ＜0.05）；冷保存24 h 后，高剂量组保存液的 ALT、MDA 含量均明显高于对照组及低、中剂量组（均为 P ＜0.05）。病理检查结果显示，高、中、低剂量组大鼠肝脏的损伤明显轻于对照组，各剂量组之间比较无明显差别。结论在 UW 液中加入 CP 对大鼠离体肝脏冷保存有较好的保护作用，优于单纯应用 UW 液保存。%Objective To discuss the protective effect of creatine phosphate (CP)on isolated rat liver against cold preservation.Methods Isolated perfused rat liver model under simple cold preservation was established.The liver of the control group was perfused with pure University of Wisconsin (UW)solution. With UW solution as the base fluid,the liver of the low-dose group was perfused with 1 g/100 ml CP in UW solution;the liver of the middle-dose group was perfused with 2 g/100 ml CP in UW solution;the liver of the high-dose group was perfused with 3 g/100 ml CP in UW solution.The livers of each group were cold preserved in the corresponding perfusion fluid at 4 ℃.The content of alanine aminotransferase (ALT)and lactate dehydrogenase (LDH)in preservation solution in infrahepatic vena cava were determined.The content of malondialdehyde (MDA)and activity of myeloperoxidase (MPO)in liver tissues were detected.The apoptosis index (AI)of liver cells in liver tissues and positive expression rate of NF-κB in liver tissues were observed. Pathologic changes of liver tissues were observed under optical microscope.Results At 12 h after the cold preservation,the content of ALT and LDH in the rat livers of low-,middle-and high-dose groups were lower than those of the control group (all in P <0.05).At 18 h after the cold preservation,the content of MDA and MPO in the liver tissues of low-,middle-and high-dose groups were lower than those of the control group (all in P <0.05).At 12 h and 18 h after the cold preservation,AI and positive expression rate of NF-κB in liver cells in the rat livers of low-,middle-and high-dose groups were lower than those of the control group (all in P<0.05).At 24 h after the cold preservation,the content of ALT and MDA in preservation solution of the high-dose group was obviously higher than that of the control group as well as the low-and middle-dose groups (all in P <0.05).The results of pathological examination indicated that the injuries to the livers of the high-,middle-and low-dose groups were obviously lighter than that of the control group.There was no obvious difference among each dose group.Conclusions CP in UW solution may well protect the isolated rat liver against cold preservation,which is better than pure UW solution.