Zuo Yuan-Yi, Zhao Yan-Bo, Jiang Xiao-Gang, Gu Zhen-Lun, Guo Ci-Yi, Bian Shi-Zhong
Department of Forensic Medicine, Medical College of Soochow University, Suzhou 215123, China.
Fa Yi Xue Za Zhi. 2011 Dec;27(6):405-8, 412.
To explore the effect of ketamine on adrenal pheochromocytoma (PC12) cell proliferation inhibition and induction of apoptosis and its mechanism.
PC12 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 72h. Hoechst stain was used to observe the morphological changes of apoptosis. PC12 cells cultured after 48 h with different concentrations of ketamine were selected to detect apoptotic rate using flow cytometry and detect the expression of bax and bcl-2 proteins using Western blotting.
For different concentrations of ketamine, vitality of PC12 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.
Ketamine can inhibit the proliferation of PC12 cell by inducing apoptosis of the PC12 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.
探讨氯胺酮对肾上腺嗜铬细胞瘤(PC12)细胞增殖抑制及诱导凋亡的作用及其机制。
以大鼠PC12细胞作为多巴胺能神经元模型。PC12细胞分别用浓度为0.9、1.2、1.5、1.8和2.1 mmol/L的氯胺酮培养。在培养12、24、48和72小时后,采用MTT法测定细胞活力。用Hoechst染色观察细胞凋亡的形态学变化。选取用不同浓度氯胺酮培养48小时后的PC12细胞,采用流式细胞术检测凋亡率,并用蛋白质免疫印迹法检测bax和bcl-2蛋白的表达。
对于不同浓度的氯胺酮,PC12细胞活力随培养时间的增加而显著降低。用Hoechst染色明显观察到细胞凋亡。流式细胞术显示,凋亡率随氯胺酮浓度的增加而显著升高。
氯胺酮可通过浓度依赖性诱导PC12细胞凋亡来抑制其增殖。其潜在机制可能与促进细胞中bax的表达和抑制bcl-2的表达有关。