CCK-8通过上调Bcl-2/Bax的比率和下调Caspase-3的表达降低谷氨酸诱导的皮层神经元凋亡  

CCK-8 prevents glutamate-induced apoptosis in cultured cortical neurons via up-regulation of bcl-2/bax ratio and down-regulation of caspase-3

作　　者：张会春[1] 周江堡[1] 

机构地区：[1]重庆医科大学附属儿童医院神经内科,重庆400014

出　　处：《重庆医科大学学报》2010年第8期1141-1144,共4页Journal of Chongqing Medical University

摘　　要：目的:研究八肽胆囊收缩素(Cholecystokinin octapeptide,CCK-8)抗谷氨酸诱导皮层神经元凋亡的相关机制。方法:体外培养皮层神经元至第8d,随机分为对照组、谷氨酸组和CCK组。对照组不予任何处理;谷氨酸组用50μmol/L谷氨酸处理30min后,换用正常培养基继续培养;CCK组在谷氨酸处理前先用CCK-8处理24h;各组细胞在谷氨酸损伤后用原培养基继续培养0、6、12、24、48h后分别获取标本待检。用噻唑蓝(MTT)染色法检测神经元活力,免疫细胞化学技术检测神经元Bcl-2、Bax和Caspase-3的表达。结果:CCK组神经元活力、Bcl-2表达较谷氨酸组增高,Bax、Caspase-3较谷氨酸组降低,差异均具有统计学意义(P<0.05)。结论:CCK-8抗谷氨酸诱导的皮层神经元凋亡的作用机制可能与上调Bcl-2/Bax比率、抑制Caspase-3的激活有关。Objective： To investigate the mechanisms responsible for the neuroprotection by cholecystokinin octapeptide against glutamate-induced apoptosis in vitro cultured cortical neurons. Methods： Primary cultured corticaI neurons from SD rats of 0～24 hold were incubated for 8 days. The cultured cells were divided randomly into three groups： control group,glutamate group and CCK group. In the controI group,cells were not treated with glutamate orCCK;Neurons in glutamate group were incubated with 50μmol/Lglutamate for 30 min;In CCK group,CCK-8 was added to the Neurons 24 h prior to incubation with glutamate. After injuried by glutamate,cells in all the groups were incubated with normal medium for 0,6,12,24 h and 48 h. At the five time points,cells were fixed respectively for experiment. Cell viability were determined by the colorimetric MTT assay;The protein expression of Bcl-2,Bax and Caspase-3 were determined by immunocytochemistry techniques. Results：Pretreatment with CCK for 24 h significantly improved glutamate-induced suppression of cell viability. Pretreatment with CCK also completely reversed the suppression of Bcl-2 expression,and significantly inhibited Bax overexpression and Caspase-3 activition induced by glutamate. Conclusion：Theneuroprotective mechanisms of CCK against glutamate-induced apoptosis in cultured cortical neurons may be associated with up-regulation of Bcl-2/Bax ratio and down-regulation of Caspase-3.

关 键 词：八肽胆囊收缩素 谷氨酸 凋亡 Bcl-2 BAX Caspase-3 

分 类 号：R589.2[医药卫生—内分泌]