瑞芬太尼预先给药对大鼠肝脏缺血再灌注损伤时肝细胞线粒体的影响

目的 探讨瑞芬太尼预先给药对大鼠肝脏缺血再灌注损伤时肝细胞线粒体的影响.方法 健康雄性SD大鼠60只,体重250～270 g,随机分为6组(n=10):对照组(C组)麻醉后直接取肝脏;缺血再灌注组(IR组)静脉输注等容量(7 ml)生理盐水;低浓度瑞芬太尼组(Rl组)静脉输注瑞芬太尼0.2 μg·kg-1·min-1;高浓度瑞芬太尼组(Rh组)静脉输注瑞芬太尼1 μg·kg-1·min-1;5-羟癸酸盐组(5-HD组)静脉输注5-HD 150 μg·kg-1·min-1;5-羟癸酸盐+高浓度瑞芬太尼组(5-HD+Rh组)静脉输注5-HD 150 μg·kg-1·min-1和瑞芬太尼1 μg·kg-1·min-1.除C组外,其余组均于输注20 min开腹,夹闭肝门30 min,再灌注60 min.观察肝细胞线粒体超微结构,制备肝细胞线粒体悬液,测定线粒体基质钙浓度(Ca2+]m)、线粒体摄钙后浓度(Ca2+]upost)、线粒体摄钙量(Ca2+]u)及丙二醛(MDA)含量.结果 C组线粒体Ca2+]m、Ca2+]upost、ca2+]u及MDA含量低于其余各组;与IR组比较,Rl组与Rh组线粒体Ca2+]m、Ca2+]upost、Ca2+]u降低;与RI组比较,Rh组Ca2+]m、Ca2+]upost降低;与Rh组比较,5-HD+Rh组和5-HD组Ca2+]m、Ca2+]upost、Ca2+]u升高(P＜0.05或0.01);5-HD组和5-HD+Rh组上述各指标差异无统计学意义(P＞0.05).结论 瑞芬太尼0.2～1 μg·kg-1·min-1预先给药可减轻大鼠肝脏缺血再灌注时肝细胞线粒体损伤,其机制可能与开放线粒体ATP敏感性钾通道有关.

The protective effects of remifentanil pretreatment on hepatocyte mitochondrial function against ischemia-reperfusion injury in rats

Objective To investigate the protective effects of remifentanil pretreatment on hepatocyte mitochondrial function against ischemia-reperfusion(I/R)injury and the underlying mechanism.Methods Sixty healthy SD rats weighing 250-270 g were randomly divided into 6 groups(n=10 each):group Ⅰ blank control (C);group Ⅱ I/R;group Ⅲ low-Remi + I/R;group Ⅳ high-Remi + I/R;group Ⅴ 5-HD + I/R and group Ⅵ 5-HD + high-Remi + I/R.The animals were anesthetized with intraperitoneal 0.5% pentobarbital 40 mg/kg.In group Ⅱ-Ⅵ the hilum of the liver was occluded for 30 min followed by 60 min reperfusion.The animals received 7 ml ofnormal saline(in group Ⅱ),remifentanil(in group Ⅲ,Ⅳ),5-HD(in group Ⅴ)and 5-HD + remifentanil livers were removed at the end of 60 min reperfusion.The mitochondria were isolated by differential centrifugation and suspended in 0.01 mol/L Tris/HCl buffer(pH 7.4).Protein concentration was determined by Bradford method.Mitochondrial Matrix calcium concentration(Ca2+]m),calcium concentration after calcium uptake (Ca2+]upost),quantity of calcium uptake(Ca2+]u) and MDA content were determined.All of the data were analyzed with SPSS 12.0 and the differences between the groups were tested by analysis of variance.Results MitochondrialCa2+]m,Ca2+]upost,Ca2+]u and MDA content were significantly higher in group Ⅱ-Ⅵ than in group Ⅰ(control group).Ca2+]m,Ca2+]upost and ca2+]u were significantly decreased in low-Remi and high-Remi groups(group Ⅲ and Ⅳ)as compared with I/R group(group Ⅱ).Ca2+]m andCa2+]upost were significantly lower in high-Remi group (Ⅳ) than in low-Remi(Ⅲ)group.Ca2+]m,Ca2+]upost andCa2+]u were sisnificantly higher in 5-HD(Ⅴ)and 5-HD + high-Remi(Ⅵ)groups than in high-Remi(Ⅳ)group.There was no significant difference in mitochondrialCa2+]m,Ca2+]upost,Ca2+]u and MDA content between can protect hepatocyte mitochondrial function against I/R injury by activating KATP channel.