氯胺酮对PC12细胞增殖及凋亡的影响

摘要

目的 探讨氯胺酮对肾上腺嗜铬细胞瘤(pheochromocytoma,PC12)细胞的增殖抑制和诱导凋亡的作用及其机制. 方法 以大鼠PC12细胞为多巴胺能神经元的细胞模型,分别以0.9、1.2、1.5、1.8、2.1 mmol/L浓度的氯胺酮加入培养的PC12细胞中,分别培养12、24、48、72h后,用MTT实验检测细胞的活力,Hoechst 染色观察其凋亡的形态,用流式细胞仪检测不同浓度的氯胺酮加入PC12细胞培养48h后细胞凋亡率以及用Western印迹法检测bax、bcl-2的蛋白表达.结果 加入不同浓度的氯胺酮,随着培养时间的延长,PC12细胞的活力显著下降,Hoechst染色可观察到明显凋亡现象,流式细胞仪检测显示随着氯胺酮浓度的增加,细胞凋亡率显著增加.结论 氯胺酮可抑制PC12细胞增殖,并可显著诱导PC12细胞凋亡,且呈浓度依赖性,其作用机制可能与促进细胞内bax以及抑制bcl-2的表达有关.%Objective To explore the effect of ketamine on adrenal pheochromocytoma(PC12) cell proliferation inhibition and induction of apoptosis and its mechanism. Methods PC 12 cells of rats were models for dopaminergic neuron. PC12 cells were cultured with ketamine at concentrations of 0.9, 1.2, 1.5, 1.8 and 2.1 mmol/L, respectively. The cell viability was measured by MTT method after incubation at 12, 24, 48 and 72 h. Hoechst stain was used to observe the morphological changes of apoptosis. PC 12 cells cultured after 48 h with different concentrations of ketamine were selected to detect apoptotic rate using flow cy-tometry and detect the expression of bax and bcl-2 proteins using Western blotting. Results For different concentrations of ketamine, vitality of PC 12 cells significantly decreased with increase of the incubation time. Apoptosis was obviously observed using Hoechst staining. Flow cytometry showed that apoptosis rates significantly increased with increasing ketamine concentrations. Conclusion Ketamine can inhibit the proliferation of PC12 cell by inducing apoptosis of the PC 12 cell in a concentrations-dependent manner. The underlying mechanism may be related to promoting the expression of bax and inhibiting the expression of bcl-2 in the cells.