依托咪酯对猪肾上腺皮质细胞的毒性作用及依托咪酯预处理对其的影响

目的 探讨依托咪酯对猪肾上腺皮质细胞的毒性作用及依托咪酯预处理对其的影响.方法 实验分两部分,第一部分为细胞毒性实验,猪肾上腺皮质细胞随机分为8组,每组3皿,对照组加入0.5%二甲基亚砜(DMSO),其他7组分别加入50、100、200、300、400、500、1000 μmol/L依托咪酯,每组分别作用6、12、24 h,进行下述指标检测.第二部分为预处理实验,猪肾上腺皮质细胞随机分为3组,每组3皿,对照组加入0.5%DMSO,损伤组加入325 μmol/L依托咪酯作用24 h,预处理组先加入0.6 μmol/L依托咪酯作用1 h,洗脱后在培养基中孵育4 h,再加入325 μmol/L依托咪酯作用24 h.采用CCK-8法检测细胞活力,计算24 h时依托咪酯半数抑制浓度(IC50),采用流式细胞仪检测细胞凋亡情况.结果 依托咪酯可抑制猪肾上腺皮质细胞活力,诱导细胞凋亡,呈浓度和时间依赖性,24 h时IC50为325μmol/L.0.6 μmol/L依托咪酯预处理1 h可抑制325μmol/L依托咪酯诱发的细胞凋亡具有抑制作用.结论 依托咪酯通过诱发猪肾上腺皮质细胞凋亡对其功能产生抑制作用;依托咪酯预处理可减轻药物自身引起的这种抑制作用.

Toxic effect of etomidate on porcine adrenal cortical cells and influence of etomidate preconditioning

Objective To investigate the effect of etomidate on porcine adrenal cortical cells and the influence of preconditioning with small dose etomidate. Methods Porcine adrenal cortical cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 15% fetal bovine serum at 37℃ in 5% CO2 incubator for 24 h. The concentration was 2 × 106/ml. The experiment was performed in 2 parts. In part Ⅰ the cells were exposed to 50, 100, 200, 300, 400, 500 and 1000 mol/L etomidate respectively and incubated for 6, 12and 24 h, while in control group, the cells were exposed to 0.5% dimethylsulfoxide (DMSO). Cell viability was measured using CCK-8 assay and apoptosis by flow cytometry, and the 50% inhibitory concentration (IC50) of etomidate was calculated at 24 h of incubation. In part Ⅱ the cells were exposed to 0.6 μmol/L etomidate for 1 h and were allowed to recover for 4 h after etomidate washout, then etomidate 325 μmol/L was added and the cells were incubated for 24 h. Cell viability and apoptosis were assessed and the IC50 of etomidate was calculated as in part Ⅰ .Results Etomidate inhibited viability of porcine adrenal cortical cells and induced apoptosis in a dose- and time-dependent manner. The IC50 of etomidate at 24 h of incubation was 325 μmol/L. Preconditioning with0.6 μmol/L etomidate for 1 h attenuated the apoptosis induced by etomidate 325 μmol/L. Conclusion Etomidate can inhibit cell viability and induce apoptosis in a dose- and time-dependent manner. Preconditioning with small dose etomidate has protective effect.