氯胺酮对体外培养大鼠神经干细胞增殖与凋亡的影响  

Effects of ketamine on proliferation of and apoptosis in cultured rat neural stem cells

作　　者：黄河[1] 杨天德[1] 杨辉[2] 陶军[1] 李洪[1] 吕胜青[2] 

机构地区：[1]第三军医大学新桥医院麻醉科,重庆市400037 [2]第三军医大学新桥医院神经外科,重庆市400037

出　　处：《中华麻醉学杂志》2006年第8期752-755,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金重点资助课题(30130110)

摘　　要：目的探讨氯胺酮对体外培养夫鼠神经干细胞(NSC)增殖与凋亡的影响。方法采用无血清培养和单细胞克隆技术,在大鼠海马分离培养具有单细胞克隆能力的细胞群,采用免疫荧光细胞化学技术证实为NSC。将NSC以5×10~4/孔接种于96孔培养板中,分别加入浓度为0(未加氯胺酮)、5、10、20、50、100、200、500、1000 mmol·L^(-1)氯胺酮,每个浓度5孔。采用四甲基偶氮唑蓝比色法检测NSC光密度(OD),并计算生长抑制率(RI)。采用流式细胞仪测定氯胺酮浓度为0、10、100、500、1000 mmol·L^(-1)时NSC凋亡率。结果与氯胺酮浓度0时比较,200、500、1000 mmol/L氯胺酮可降低NSC OD,升高RI,10、100、500、1000 mmol·L^(-1)氯胺酮可升高NSC凋亡率(P＜0．05或0．01)。结论氯胺酮对体外培养大鼠NSC增殖有抑制作用,并能诱导NSC凋亡。Objective To study the effects of different concentrations of ketamine on proliferation of and apoptosis in the cultured rat neural stem cells （NSCs）.Methods Single cells which had the ability to form clones were isolated from hippocampus of embryo obtaind from 14d pregnant Wistar rats using serum-free cell culture and single cell clone technique and were identified to be neural stem cells （NSCs） by immuno-fluorescence technique. Different concentrations of ketamine （0, 5, 10, 20, 50, 100, 200, 500, 1 000 mmol·L^-1） were added to the cells and incubated at 37 ℃ and in 5 % CO2 for 5 days. The effect of different concentrations of ketamine on growth inhibition rate （IR） of NSCs was calculated by determination of OD （optical density） of cultured rat NSCs using MTT assay. IR = OD in control group （OD in ketamine group/OD in control group. The effect of different concentrations of ketamine （0, 10, 50, 100, 500, 1 000 mmol· L^- 1 ） on the apoptosis rate of NSCs was measured by flow cytometry. Results Ketamine （100, 200, 500, 1 000 mmol·L^-1 ） inhibited the growth of cultured NSCs. Flow cytometry showed that apoptosis rate of NSCs was significantly higher when ketamine concentration was 10, 50, 100, 500, 1 000 mmol· L^-1 than that in control group （ketamine = 0） . Conclusion Ketamine can inhibit proliferation of cultured rat NSCs and induce apoptosis.

关 键 词：氯胺酮 干细胞 细胞凋亡 细胞增殖 

分 类 号：R96[医药卫生—药理学]