CAJ | 학술논문

目的评价p38丝裂原蛋白激酶(p38MAPK)信号通路在瑞芬太尼或缺血预处理减轻大鼠肝缺血再灌注损伤中的作用.方法健康SD大鼠l44只,雌雄不拘,体重200 ～ 250 g,采用随机数字表法,将其随机分为6组(n=24):假手术组(S组)；肝缺血再灌注组(I/R组)采用动脉夹夹闭左叶和中叶肝蒂30 min,恢复灌注的方法制备大鼠肝缺血再灌注模型；瑞芬太尼组(R组)于缺血前30 min静脉输注瑞芬太尼2μg·kg-1·min-1至再灌注120 min；缺血预处理组(IPC组)于缺血前30 min行缺血5 min,再灌注5 min,重复3次后制备缺血再灌注模型；SB+R组和SB+ IPC组分别于输注瑞芬太尼或缺血预处理前5 min静脉注射p38mAPK特异性抑制剂SB203580 0.2 mg/kg,其余组给予等体积生理盐水.于再灌注30、60、90和120 min时各组分别随机取6只大鼠抽取肝下腔静脉血测定血清ALT和AST活性；采用ELISA法测定TNF-α及IL-1β浓度；随后处死大鼠,取肝组织,采用Western blot法测定磷酸化p38MAPK的表达,观察病理学结果.结果与S组相比,I/R组各时点血清ALT、AST、TNF-α及IL-1β水平升高(P＜0.05),病理学损伤明显加重；与I/R组相比,RPC组和IPC组血清ALT和AST活性、TNF-α及IL-1β浓度降低,再灌注90 min时磷酸化p38MAPK表达上调(P＜0.05),SB+ RPC组和SB+ IPC组各指标差异无统计学意义(P＞0.05)；SB+ RPC组与RPC组,SB+ IPC组和IPC组相比,血清ALT、AST、TNF-α及IL-1β水平升高,再灌注90 min时磷酸化p38MAPK表达下调(P＜0.05),病理学损伤明显加重.结论瑞芬太尼和缺血预处理可减轻大鼠肝缺血再灌注损伤,其机制可能与激活p38MAPK信号通路抑制炎性反应有关.
목적 평개p38사렬원단백격매(p38MAPK)신호통로재서분태니혹결혈예처리감경대서간결혈재관주손상중적작용.방법 건강SD대서l44지,자웅불구,체중200 ～ 250 g,채용수궤수자표법,장기수궤분위6조(n=24):가수술조(S조)；간결혈재관주조(I/R조)채용동맥협협폐좌협화중협간체30 min,회복관주적방법제비대서간결혈재관주모형；서분태니조(R조)우결혈전30 min정맥수주서분태니2μg·kg-1·min-1지재관주120 min；결혈예처리조(IPC조)우결혈전30 min행결혈5 min,재관주5 min,중복3차후제비결혈재관주모형；SB+R조화SB+ IPC조분별우수주서분태니혹결혈예처리전5 min정맥주사p38mAPK특이성억제제SB203580 0.2 mg/kg,기여조급여등체적생리염수.우재관주30、60、90화120 min시각조분별수궤취6지대서추취간하강정맥혈측정혈청ALT화AST활성；채용ELISA법측정TNF-α급IL-1β농도；수후처사대서,취간조직,채용Western blot법측정린산화p38MAPK적표체,관찰병이학결과.결과 여S조상비,I/R조각시점혈청ALT、AST、TNF-α급IL-1β수평승고(P＜0.05),병이학손상명현가중；여I/R조상비,RPC조화IPC조혈청ALT화AST활성、TNF-α급IL-1β농도강저,재관주90 min시린산화p38MAPK표체상조(P＜0.05),SB+ RPC조화SB+ IPC조각지표차이무통계학의의(P＞0.05)；SB+ RPC조여RPC조,SB+ IPC조화IPC조상비,혈청ALT、AST、TNF-α급IL-1β수평승고,재관주90 min시린산화p38MAPK표체하조(P＜0.05),병이학손상명현가중.결론 서분태니화결혈예처리가감경대서간결혈재관주손상,기궤제가능여격활p38MAPK신호통로억제염성반응유관.
Objective To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) pathway in the protective effect of remifentanil or ischemic preconditioning (IPC) against hepatic ischemia-reperfusion (I/R) injury in rats.Methods One hundred and forty-four male SD rats,weighing 200-250 g,were randomly assigned into 6 group ( n =24 each):sham operation group (group S),I/R group,remifentanil group (group R),IPC group,SB203580 (a specific p38MAPK inhibitor) + remifentanil group (group SB + R),and SB + IPC group.The animals were anesthetized with intraperitoneal 20％ urethane 1 mg/kg.Partial liver ischemia was produced by clamping the hepatic pedicle supplying left lobe and middle lobe for 30 min,followed by 120 min reperfusion.In group R,remifentanil was infused intravenously at 2μg· kg- 1 · min- 1 starting from 30 min before ischemia until the end of reperfusion.In IPC group,the rats were subjected to 3 episodes of 5 min ischemia at 5 min intervals before I/R.SB203580 0.2 mg/kg was injected intravenously at 5 min before remifentanil infusion or IPC in groups SB + R and SB + IPC,and the equal volume of normal saline was given in the other groups.Six rats in each group were selected at 30,60,90 and 120 min of reperfusion and venous blood samples were taken from inferior vena cava for measurement of serum ALT and AST activities and concentrations of TNF-a and 1L-1β.The rats were then sacrificed and liver tissues were taken for microscopic examination and determination of phosphor-p38MAPK expression by Western blot.Results Compared with group S,serum AST and ALT activities and serum levels of TNF-α and IL-1β were significantly increased at each time point (P ＜ 0.05) and pathological injury was aggravated in group I/R.Compared with group I/R,serum AST and ALT activities and serum levels of TNF-a and IL-lβ were significantly decreased and phosphor-p38MAPK expression was up-regulated at 90 min of reperfusion in groups R and IPC ( P ＜ 0.05).The serum AST and ALT activities and serum levels of TNF-α and IL-1β were significantly increased,phosphor-p38MAPK expression was down-regulated at 90 min of reperfusion ( P ＜ 0.05),and pathological injury was aggravated in group SB + R compared with group R,and in group SB + IPC compared with group IPC.Conclusion Activation of p38MAPK pathway and inhibition of inflammatory response may be involved in the mechanism by which remifentanil or IPC reduces the hepatic I/R injury in rats.