摘要目的 探讨异丙酚对大鼠肝BRL-3A细胞缝隙连接功能及顺铂诱导肝BRL-3A细胞毒性的影响.方法实验Ⅰ:将体外培养的大鼠肝BRL-3A细胞接种于12孔板内,采用随机数字表法,将其随机分为6组(n=8):对照组(C组)、脂肪乳组(10μg/ml,Ⅰ组)、油酸酰胺组(25 μmol/L,O组)和不同浓度异丙酚组[P1组(1.5 μg/ml)、P2组(2.8μg/ml)、P3组(4.1μg/ml)].采用细胞接种荧光示踪法计算荧光示踪剂传递率和抑制率.实验Ⅱ:用不同密度接种方法获得大鼠肝BRL-3A细胞的高密度组(接种密度为1×105个/ml)和低密度组(接种密度为500个/ml),采用随机数字表法,将两组各分为5个亚组(n=8):空白对照亚组;顺铂亚组加入顺铂2.5 μmol/L;顺铂+脂肪乳亚组加入顺铂2.5 μmol/L及脂肪乳10 μg/ml;顺铂+油酸酰胺亚组加入顺铂2.5 μmol/L及油酸酰胺25 μmol/L;顺铂+异丙酚亚组加入顺铂2.5 μmol/L及异丙酚2.8μg/ml,其中油酸酰胺和异丙酚均于顺铂之前加入,单独的作用时间分别为1 h和3 h,与顺铂共同的作用时间为1 h.采用标准细胞集落形成分析法评估异丙酚对顺铂的细胞毒性.结果 与C组比较,O组、P1组、P2组和P3组荧光示踪剂传递率降低,抑制率升高(P＜0.05);异丙酚呈浓度依赖性地降低荧光示踪剂传递率及升高抑制率(P＜0.05).在高密度组中,异丙酚和油酸酰胺可升高细胞集落形成率(P＜0.05),但在低密度组中,未见上述效应(P＞0.05).结论 异丙酚呈浓度依赖性地抑制大鼠肝BRL-3A细胞的缝隙连接功能;异丙酚可减弱顺铂诱导的大鼠肝BRL-3A细胞毒性,其机制可能与抑制细胞缝隙连接功能有关.

abstractsObjective To investigate the effect of propofol on the gap junction function of BRL-3A cells and cisplatin-induced toxicity to rat hepatic BRL-3A cells. Methods Rat hepatic BRL-3A cell line was provided by professor Tao laboratory of department of pharmacology of our university and cultured in DMEM liquid culture medium at 37℃. The cells were randomly divided into 6 groups: group Ⅰ control (group C) ; group Ⅱ was exposed to intralipid (the solvent) 10 μg/ml (group Ⅰ) ; group Ⅲ was exposed to oleamide (gap junction function inhibitor) 25 μmol/L (group O) and group Ⅳ ,Ⅴ , Ⅵ were exposed to propofol l.5, 2.8 and 4.1 μg/ml respectively (groups P1, 2,3) . Using "parachute" dye-coupling assay, the dye spread rate and inhibition rate were calculated to measure the gap junction function of BRL-3 A cells. BRL-3 A cells were seeded in two different densities:high density group (seeding density = 1×105 /ml) and low density group (seeding density = 500/ml). Both groups were further divided into 5 subgroups: subgroup Ⅰ control; subgroup Ⅱ was exposed to cisplatin 2.5 μmol/L and subgroup Ⅲ ,Ⅳ ,Ⅴ were exposed to cisplatin 2.5 μmol/L + intralipid 10 μg/ml or oleamide 25 μmol/L or propofol 2.8 μg/ml respectively. The effect of propofol on the cytotoxicity of cisplatin was evaluated by standard colony-forming assay. Results The dye spread rate was significantly lower and inhibition rate higher in groups O,P1 , P2 and P3 than in group C. Propofol decreased the dye spread rate and increased inhibition rate in a concentration-dependent manner. Propofol and oleamide significantly increased colony formation rate in high density group,but had no significant effect on colony formation rate in low density group. Conclusion Propofol can inhibit the gap junction function of BRL-3A cells in a concentration-dependent manner and reduce cisplatin-induced cytotoxicity by inhibiting gap junction function.