丙泊酚对谷氨酸诱导的PC12细胞凋亡的影响

Effect of propofol on the apoptosis of PC12 cell induced by glutamic acid

目的探讨丙泊酚对谷氨酸诱导的PC12细胞凋亡的影响。方法用10 mmol·L^(-1)谷氨酸损伤PC12细胞48 h,并分为模型组和丙泊酚低、中、高剂量组。取正常生长的PC12细胞作为对照组。对照组和模型组予以不含药物培养基培养48 h,丙泊酚低、中、高剂量组分别予以12.5,25.0,50.0μmol·L^(-1)丙泊酚培养48 h。通过噻唑蓝(MTT)法检测细胞活力,用流式细胞术检测细胞凋亡,用紫外分光光度法检测Caspase-3活性,用反转录聚合酶链式反应法和Western blot检测即刻早基因(c-fos)与早期生长反应因子基因-1(Egr-1)mRNA及蛋白水平。结果与对照组比较,模型组的细胞活力显著下降(P<0.01),凋亡率显著提高(P<0.01),Caspase-3活性显著上升(P<0.01),c-fos mRNA及蛋白水平显著下调(P<0.01),Egr-1 mRNA及蛋白水平显著上调(P<0.01)。与模型组比较,丙泊酚低、中、高剂量组的细胞活力显著提高(P<0.01),细胞凋亡率显著下降(P<0.01),Caspase-3活性显著降低(P<0.01),c-fos mRNA及蛋白水平显著上调(P<0.01),Egr-1 mRNA及蛋白水平显著下调(P<0.01)。结论丙泊酚可显著抑制谷氨酸诱导的PC12细胞凋亡,其作用机制与调节c-fos及Egr-1的表达有关。

Objective To explore the effect of propofol on the apoptosis of PC12 cell induced by glutamie acid. Methods PC12 cells were in- duced 10 mmol . L-1 glutamate for 48 h, and were divided into model group, propofol low, medium and high dose groups. The normal cells were used as control group. Control group and model group were received culture medium without any drugs for 48 h. Propofol low, medium and high dose groups were given 12. 5, 25.0, 50. 0 μmol . L-1 propofol for 48 h. The viability of PC12 cell was measured by MTT assay. PC12 cell apoptosis was measured by flow cytometry. The activity of Caspase -3 was determined by spectrophotometric method. The expression of FBJ osteosareoma oncogene （c -fos ） and early growth response protein 1 （Egr- 1 ） were detected by Realtime -PCR and western blot. Results Compared with the normal group, the cell apoptosis rate, the activity of Caspase - 3, the expression of c - los mRNA and protein increased, and the cell viability, the expression of Egr - 1 mRNA and protein decreased in the model group （ P 〈 0. 01 ）. Compared with the model group, the acitivity of Caspase- 3, the cell apoptosis rate, the activity of Caspase - 3, the expression of c - fos mRNA and protein decreased, and the ceU viability, the expression of Egr- i mRNA and protein increased in propofol low- dose, medium- dose and high -dose group （P 〈 0. 05 ）. Conclusion Propofol suppressed the apoptosis of PC12 cell induced by glutamic acid, which was related with the expression of c - fos and Egr - 1.