不同浓度瑞芬太尼预处理对人肝细胞缺氧复氧损伤保护作用的影响

Effect of Remifentanil Preconditioning in Different Concentrations on Hepatic Hypoxia-reoxygenation Injury

目的研究不同浓度瑞芬太尼预处理对人肝细胞缺氧复氧损伤保护作用的影响,探讨瑞芬太尼预处理的适宜浓度。方法将培养的人肝细胞按瑞芬太尼的浓度梯度分为11组：正常对照组（N组）,缺氧复氧组（IR组）,瑞芬太尼预处理组（RE1~RE8组）：瑞芬太尼终浓度分别为0.5,1,5,10,20,30,40,50 ng.mL 1预处理1 h后建立缺氧复氧模型,生理盐水组（NS组）。N组正常条件下培养,其他组缺氧8 h,复氧4 h。复氧结束即刻,MTT法检测肝细胞活力;全自动生化仪检测细胞培养液中谷草转氨酶（AST）、谷丙转氨酶（ALT）和乳酸脱氢酶（LDH）水平;MDA试剂盒检测肝细胞内丙二醛（MDA）含量。结果与N组比较,其他各组细胞活力降低,AST,LDH水平和细胞内MDA含量升高（P〈0.05）。与IR组和NS组比较,RE2~RE6组细胞活力升高,AST、LDH水平和细胞内MDA含量降低（P〈0.05）;而RE1、RE7和RE8组各项指标差异无统计学意义（P〉0.05）。结论临床有效血药浓度（1~30 ng.mL 1）瑞芬太尼预处理对肝细胞缺氧复氧损伤具有保护作用,而低浓度（0.5 ng.mL 1）和高浓度（≥40 ng.mL 1）瑞芬太尼预处理则无此作用。

OBJECTIVE To investigate different remifentanil concentrations used to pretreat hepatocytes undergone hypoxia-reoxygenation injury to find out the proper concentration against hypoxia-reoxygenation injury. METHODS Hepatocytes HL7702 were cultured in vitro, and invided into eleven groups： control group（group N）, hypoxia-reoxygenation group（group IR）, remifentanil preconditioning groups （group RE1-RE8） and normal saline group（group NS） were administered one hour before hepatocyte hypoxia-reoxygenation injury at different concentrations of 0.5, 1, 5, 10, 20, 30, 40, 50 ng.mL-1 and normal saline respectively. Then 8 hours of hypoxia and 4 hours of reoxygenation were followed in the group IR, group NS and the preconditioning groups, while the group N was cultured in normal condition. Cell viability, ALT, AST, LDH and intracellular MDA were measured. RESULTS Compared with group N, cell viability reduced, AST, LDH and intracellular MDA increased in other groups （P〈0.05）. Compared with group IR and group NS respectively, cell viability increased, AST, LDH and intracellular MDA reduced in groups RE2-RE6 （P〈0.05）, however, no differences were shown among group RE1, RE7 and RE8 （P〉0.05）. CONCLUSION Remifentanil preconditioning protects against hypoxia-reoxygenation injury in human hepatocyte. The range of effective concentration is among 1-30 ng.mL-1, which is in accordance with the clinically effective concentration in anesthesia, while the concentration out of the range shows no protection.