炎症性细胞因子致肝脏缺血再灌注损伤和丹参单体的保护作用

目的：探讨肿瘤坏死因子α（TNF-α）和白细胞介素8（IL－8）在肝脏缺血再灌注损伤中的变化以及水溶性丹参单体MP－1（简称MP－1）的作用。方法：建立人肝窦内皮细胞体外低温缺氧再氧化模型和大鼠离体肝脏缺血再灌注损伤模型，以台盼蓝染色观察细胞损伤情况，用放免法分析透明质酸吸收率来反映肝AST、LDH的变化反映肝脏功能。结果：低温缺氧再氧化期间，肝窦内皮细胞的死亡率随着时间的增加而增加，TNF-α和IL－8的释放也随之增加，但肝窦内皮细胞的功能下降；肝窦内皮细胞死亡率分别与TNF-α与IL－8的释放呈正相关（r=0.949,P<0.05和r=0.892,P<0.05)。加入TNF-α抗体低温缺氧再氧化6h的内皮细胞死亡率降低；重组TNF-α处理的肝窦内皮细胞死亡率明显增加；离体大鼠肝脏的功能随低温保存和再灌注时间的增加而降低；MP－1能明显降低人肝窦内皮细胞的死亡率及TNF-α和IL－8的释放，同时可以减轻离体大鼠肝脏的缺血再灌注损伤。结论：TNF-α直接参与肝窦内皮细胞的缺血再灌注损伤，MP－1可能通过抑制TNF-α和IL－8的途径来减轻肝脏缺血再灌注损伤。

Protective Effect of Hydrophilic Salvia Monomer on Liver Ischemia/Reperfusion Injury Induced by Pro-inflammatory Cytokines

OBJECTIVE: To observe the changes of tumor necrosis factor alpha (TNF alpha), interleukin 8 (IL-8) in liver ischemia/reperfusion injury and the protective effect of hydrophilic Salvia monomer MP-1 on them. METHODS: Hypothermic hypoxia reoxygenation model of human liver sinusoidal endothelial cell line and ischemia/reperfusion model of isolated rat liver were used. TNF alpha and IL-8 were measured with ELISA kits. Cell injury was excluded by trypan blue stain, sinusoidal endothelial cell function was assessed by hyaluronic acid uptake rate through RIA. Liver function was assayed by alanine transaminase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) release as well as output of bile flow. RESULTS: During hypoxia reoxygenation, sinusoidal endothelial cell injury, TNF alpha and IL-8 increased significantly in time-dependent manner, while sinusoidal cell function decreased. Cell injury was positively correlated to the released amount of TNF alpha (r = 0.949, P < 0.05) and IL-8 (r = 0.892, P < 0.05), respectively, the mortality could be reduced within 6 hrs by adding anti-TNF alpha monoclonal antibody and increased by treating with recombinant human TNF alpha. Function of isolated rat liver lowered alone the increasing of low temperature ischemia/reperfusion time. MP-1 could markedly lower the mortality of endothelial cells and TNF alpha and IL-8 release, it also could alleviate ischemia/reperfusion injury to isolated rat liver. CONCLUSION: TNF alpha participated in liver ischemia/reperfusion injury directly, and MP-1 might alleviate the injury through inhibiting TNF alpha and IL-8.