可卡因诱导胎鼠大脑皮层神经元细胞凋亡探究.docVIP

可卡因诱导胎鼠大脑皮层神经元细胞凋亡探究 　 作者：吴岚莹，王华， 沈朝斌，戴强，孙金平，顾君，夏尤佳，龚建宏;【摘要】; 目的; 探讨使用不同剂量可卡因对大脑皮层神经元细胞凋亡的。; 实验组每日分别给妊娠Wistar大鼠皮下注射不同剂量的盐酸可卡因15天。对照组每日给予相同剂量的普通生理盐水15天。在可卡因使用的第15天分别取胎鼠大脑皮层组织，用原位缺口末端标记（TUNEL）法、流式细胞术（FCM）检测大脑皮层神经元细胞凋亡。结果; 在使用可卡因15mg/kg和25mg/kg组出现细胞凋亡，同对照组相比差异有非常显著性（Plt;0.01），而1.5mg/kg组未出现细胞凋亡。结论; 使用大剂量可卡因可以导致胎鼠大脑皮层神经元细胞凋亡。 ; 【关键词】; 可卡因; 神经元; 凋亡 ; Cocaine induced cell apoptosis in cortical neurons of fetal mice ; 【Abstract】; Objective; To investigate the role of cell apoptosis induced by cocaine with different doses.Methods; Pregnant Wistar rats in experimental group were given cocaine hydrochloride with different doses according to body weight(Subcutaneously injected for 15 days). Control animals received physiological saline of equal volumes for 15 days. Cortical tissues of fetal mice were removed after 15 days in two groups. TUNEL (In situ nick end-labelling) assay was carried out to detect cell apoptosis， FCM (flow -cytometry）assay was carried out to detect cell apoptosis in cortical neurons of fetal mice.Results; number of apoptotic cells was significantly increased in cocaine exposure at doses of 15mg/kg and 25mg/kg ，it were significantly different than the controls (Plt;0.01).But that at a dose of 1.5mg/kg did not expressed apoptosis.Conclusion; The experimental results suggest that cocaine exposure; at big doses lead to significant apoptosis in cortical neurons of fetal mice. ; 【Key words】; cocaine; neuron; apoptosis ; 据报道在大多数北美的城市中有6%～15%的孕妇可卡因成瘾，近年来国内一些地区也存在孕妇吸毒。虽然可卡因对胚胎大脑的致畸作用已充分阐明，但对神经发育过程中出现的混乱仍存在争议。笔者通过动物实验，探讨使用可卡因对胎鼠大脑皮层神经元细胞的损害。 ; 1; 材料与方法 ; 1.1; 动物分组; 将妊娠Wistar大鼠20只，置于室温（20±2）℃的清洁级动物室中，自由进食饮水。将其分成两组：实验组和对照组。实验组共15只妊娠Wistar大鼠，分实验组A、实验组B、实验组C，每组5只妊娠Wistar大鼠。给实验组A、B、C每组大鼠分别皮下注射溶解于0.2ml蒸馏水中盐酸可卡因（青海制药厂）1.5mg/kg、15mg/kg、25mg/kg，后两者为重度可卡因服用者的剂量［1］。每天给每只大鼠称重1次，根据体重调整盐酸可卡因的剂量。对照组5只妊娠Wistar大鼠每天给0.2ml普通生理盐水。此处理持续15天。从每只妊娠Wistar大鼠中分别取出存活的胎鼠3只（死亡除外）。并取大脑皮层组织做检测。每组胎鼠各15只。 ; 1.2; 原位缺口末端标记（TUNEL）法检测细胞凋亡; 各组所取组织用10%中性缓冲福尔马林固定，常规石蜡切片。原位细胞凋亡检测试剂盒（POD）购自德国BOCKINGER MA