地氟醚预处理对人脐静脉内皮细胞缺氧/复氧损伤的影响

目的 探讨地氟醚预处理对人脐静脉内皮细胞缺氧/复氧损伤的影响.方法 人脐静脉内皮细胞株接种于培养板或培养皿中,随机分为5组(n=5),正常对照组(C组)不行任何处理;缺氧/复氧组(A/R组)缺氧30 min后,复氧60 min;缺氧/复氧+炎症介质刺激组[A/R+重组人肿瘤坏死因子-α(rhTNF-α)组]在复氧同时加入10 ng/ml rhTNF-α 10 μl;地氟醚预处理+缺氧/复氧组(Des+A/R组)及地氟醚预处理+缺氧/复氧+炎症介质刺激组(Des+A/R+ rhTNF-α组)先给予7.2%地氟醚30 min,洗脱10 min,然后进行缺氧/复氧,复氧同时加入10 ng/ml rhTNF-α 10 μl.采用流式细胞术和原位缺口末端标记法测定细胞凋亡情况,计算细胞凋亡率;应用透射电镜观察细胞凋亡和坏死情况.结果 与C组比较,A/R组和A/R+rhTNF-α组细胞凋亡率升高(P＜0.05);与A/R组比较,Des+A/R组细胞凋亡率降低(P＜0.05);与A/R+rhTNF-α组比较,Des+A/R+rhTNF-α组细胞凋亡率降低(P＜0.05).电镜下,A/R组和A/R+rhTNF-α组可见凋亡和坏死细胞,Des+A/R组和Des+A/R+rhTNF-α组细胞处于增殖和修复状态.结论 7.2%地氟醚预处理30 min可减轻人脐静脉内皮细胞缺氧/复氧损伤.

Effect of desflurane preconditioning on human umbilical vein endothelial cells against anoxia-reoxygenation injury

Objective To investigate the protective effect of desflurane preconditioning on human umbilical vein endothelial cells against anoxia-reoxygenation(A/R)injury.Methods The human umbilical vein endothelial cell line(ECV304)was provided by the Liver Cancer Institute,Zhongshan Hospital,Fudan University.ECV304 cells were randomly divided into 5 groups:group Ⅰ normal control;group Ⅱ A/R;group Ⅲ A/R+rhTNF-α;group Ⅳ Des + A/R and group Ⅴ Des + A/R + rhTNF-α.In group Ⅱ-Ⅴ the cells were exposed to 95% N2 + 5% CO2 in an incubator for 30 min followed by 60 min reoxygenation.In group Ⅲ and Ⅴ rhTNF-α(10 ng/ml)10 μl was added to the cells as soon as reoxygenation was started,while in group Ⅳ and Ⅴ the cells were pretreated with 7.2% desflurane for 30 min followed by 10 min washout before A/R.Apoptosis in endothelial cells was detected by fluorescence flow cytometry and TUNEL.Endothelial cells were examined with electron microscope for apoptosis and necrosis.Results The rates of apoptosis in the endothelial cells were significantly higher in A/R group and A/R + rhTNF-α group than in control group.Desflurane preconditioning significantly attenuated apoptosis in the endothelial cells induced by A/R and A/R + rhTNF-α respectively.Electron microscopy showed that there were significantly more necrotic cells in A/R group and A/R + rhTNF-α group.However in the two desflurane preconditioning groups(Ⅳ and Ⅴ)the cells were in a state of duplication and self-repairing.Conclusion Preconditioning with 30 min 7.2% desflurane can attenuate the A/R-induced injury to human umbilical vein endothelial cells.