【作者】 王冠；

【导师】 熊君宇；

【作者基本信息】 大连医科大学 ， 麻醉学， 2008， 硕士

【摘要】 目的:以急性髓性细胞白血病细胞HL-60细胞凋亡作为研究对象,在已明确预处理的给药方式可以降低静脉麻醉药物免疫抑制作用的基础上,进一步研究依托咪酯预处理对HL-60细胞线粒体膜电位改变和线粒体通透转换孔开放的影响。方法:正常培养HL-60细胞,浓度调节至1.5×10~5 /ml,按照不同的处理方法进行分组:1.对照组:仅在无依托咪酯的正常培养基中培养24h;2.预处理组:用含1μM依托咪酯的培养基预处理1h,洗去依托咪酯后在培养基中恢复4h,再用含500μM依托咪酯的培养基刺激24h;3.损伤组:仅用含500μM依托咪酯的培养基刺激24h。收集细胞后,分别用CCK-8法检测细胞活性,荧光探针JC—1检测细胞线粒体膜电位的改变,荧光探针Calcein AM荧光染色检测线粒体通透转换孔开放的程度。结果:1.CCK-8实验结果表明,与对照组相比,预处理组和损伤组的细胞生存率均有下降;但与损伤组相比,预处理组的细胞生存率增高55.57%,具有显著性差异,P < 0.01;2.JC-1检测细胞线粒体膜电位改变的实验结果显示,与损伤组相比,预处理组绿色荧光线粒体细胞所占比例平均下降25.34%;3.检测线粒体通透转换孔开放的试验中,对照组与预处理组钙黄绿素平均荧光强度值变化(管2-管3)分别为33.71和49.38,预处理组较对照组荧光强度值变化升高15.68,证明预处理组细胞线粒体通透转换孔开放受到抑制。结论:小剂量依托咪酯预处理可以减轻并延迟线粒体膜电位的下降,其机制与线粒体通透转换孔活性受抑制有关。更多还原

【Abstract】 Objective: Choose the apoptosis of HL-60 cells as the research object. Through the former study, the administration method of precondition may decrease the immunosuppressive effect of intravenous anesthetics, on the basis of which, the effects of etomidate preconditioning on the change of mitochondrial membrane potential and the opening of the mitochondrial permeability transition Pore in HL-60 Cells are further studied.Methods: HL-60 cells were cultured at a concentration of 1.5×10~5/ml. Three groups were processed in accordance to different methods. The control group was blank (0μM etomidate for 24h); the preconditioning group was precondictioned with 1μM etomidate for 1h, washed etomidate out and allowed to recover for 4h, and then treated with 500μM for 24h; the injury group was treated with 500μM etomidate for 24h. CCK-8 assay measured the cell viability and proliferation. Using the fluorescence probe of JC-1 to detect the change of mitochondrial membrane potential and the fluorescence probe of Calcein AM to detect the opening of the mitochondrial permeability transition Pore.Results: 1.In CCK-8 assay, etomidate inhibited HL-60 viability in preconditioning and injury groups, but the percentage of active cells increased by 55.57% in preconditioning group compared with that in the injury group. 2. In the detection of mitochondrial membrane potential by fluorescent JC-1, the depressed proportion of mitochondrial membrane potential was decreased by 25.34%. 3. In the detection of the mitochondrial permeability transition pore opening, the fluorescence intensity was increased by 28.69% in the preconditioning group compared with that of the control group, which indicated that the opening of the mitochondrial permeability transition pore in the precondition group was inhibited.Conclusions: Precondition at low concentration of etomidate can reduce and delay the decrease of the mitochondrial membrane potential, the mechanism of which may be relevant to the inhibition of the mitochondrial permeability transition pore.更多还原