外源性八肽胆囊收缩素对谷氨酸诱导大鼠大脑皮层神经元凋亡的保护作用研究  

Protective effect of exogenous cholecystokinin octapeptide on glutamate-induced neuronal apoptosis in the rat cerebral cortex

作　　者：王衍[1] 陈玲[2] 庞胤[3] 张海峰[3] 李贺扬 WANG Yan;CHEN Ling;PANG Yin;ZHANG Haifeng;LI Heyang(Department of Medicine,Cangzhou Medical College,Cangzhou 061001,China;Department of Gynaecology,People’s Hospital Affiliated to Cangzhou Medical College,Cangzhou 061001;Department of Basic Medicine,Cangzhou Medical College,Cangzhou 061001;Department of Neurology V,Cangzhou Central Hospital,Cangzhou 061001)

机构地区：[1]沧州医学高等专科学校医学系,河北沧州061001 [2]沧州医学高等专科学校附属人民医院妇科,河北沧州061001 [3]沧州医学高等专科学校基础医学部,河北沧州061001 [4]河北沧州市中心医院神经外五科,河北沧州061001

出　　处：《中国比较医学杂志》2023年第6期62-67,74,共7页Chinese Journal of Comparative Medicine

基　　金：2021年医学科学研究课题(20210971)。

摘　　要：目的探讨外源性八肽胆囊收缩素(CCK-8)对谷氨酸诱导体外大鼠大脑皮层神经元损伤模型中神经细胞凋亡的保护作用及其可能作用机制。方法不同浓度CCK-8处理体外培养的大鼠大脑皮层神经元,MTT法检测细胞存活情况并筛选合适浓度。培养大鼠大脑皮层神经细胞并用谷氨酸(Glu)处理建立神经元损伤模型,分为模型组、CCK-8组,另设对照组。采用流式细胞术检测细胞周期与凋亡率;Flou-4实验检测各组细胞Ca^(2+)水平;实时定量PCR法检测B细胞淋巴瘤因子2(Bcl-2)、Bcl-2相关蛋白(Bax)mRNA表达情况;Western blot检测Bcl-2、Bax、cleaved caspase-3蛋白表达情况。结果与对照组比较,模型组神经元细胞存活率、G_(1)期细胞分布比例、Bcl-2 mRNA和蛋白表达水平降低,S期、G_(2)/M期细胞分布比例、细胞凋亡率、Ca^(2+)水平、Bax和cleaved caspase-3蛋白表达水平升高(P<0.05)。与模型组比较,CCK-8组神经元细胞存活率、G_(1)期细胞分布比例、Bcl-2 mRNA和蛋白表达水平升高,S期、G_(2)/M期细胞分布比例、细胞凋亡率、Ca^(2+)水平、Bax和cleaved caspase-3蛋白表达水平降低(P<0.05)。结论CCK-8对谷氨酸诱导的神经元细胞凋亡有一定的抑制作用,可能是通过抑制钙离子内流,调节细胞内Bax/Bcl-2表达水平发挥作用。Objective To investigate the effect and mechanism of exogenous cholecystokinin octapeptide(CCK-8)on neuronal apoptosis induced by glutamate in the rat cerebral cortex in vitro.Methods Rat cortical neurons cultured in vitro were treated with various concentrations of CCK-8.An MTT assay was used to assess cell survival and screen the appropriate concentration.Rat cortical neurons were treated with glutamic acid(Glu)to establish a neuron injury model and were divided into model,CCK-8,and control groups.The cell cycle and apoptosis rate were assessed by flow cytometry.The Ca^(2+)level was assessed by Flou-4 staining.Bcl-2 and Bax mRNA expression was assessed by Real-time quantitative PCR.Bcl-2,Bax,and cleaved caspase-3 protein expression was assessed by Western blot.Results Compared with the control group,the survival rate of neuronal cells,the distribution of G_(1)phase cells,Bcl-2 mRNA and protein expression levels were decreased,and the distributions of S and G_(2)/M phase cells,the apoptosis rate,calcium ion level,Bax and cleaved caspase-3 protein expression levels were increased in the model group(P<0.05).Compared with the model group,the neuronal cell survival rate,G_(1)phase cell distribution,snf Bcl-2 mRNA and protein expression levels were increased,whereas S and G_(2)/M phase cell distributions,apoptosis rate,calcium ion level,Bax and cleaved caspase-3 protein expression levels were decreased in the CCK-8 group(P<0.05).Conclusions CCK-8 has an inhibitory effect on neuronal apoptosis induced by glutamate by inhibiting calcium influx and regulating Bax/Bcl-2 expression in cells.

关 键 词：外源性八肽胆囊收缩素 大脑皮层神经元 细胞凋亡 B细胞淋巴瘤因子2 Bcl-2相关蛋白 

分 类 号：R-33[医药卫生]