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
作者:
作者单位:

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

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R-33


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

1. Department of Medicine, Cangzhou Medical College, Cangzhou 061001, China. 2. Department of Gynaecology, People’s Hospital Affiliated to Cangzhou Medical College, Cangzhou 061001. 3. Department of Basic Medicine, Cangzhou Medical College, Cangzhou 061001. 4. Department of Neurology V, Cangzhou Central Hospital, Cangzhou 061001

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    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Ca2+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、G1 期细胞分布比例、Bcl-2 mRNA 和蛋白表达水平降低,S 期、G2 / M 期细胞分布比例、细胞凋亡率、Ca2+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、G1 期细胞分布比例、Bcl-2 mRNA 和蛋白表达水平升高,S 期、G2 / M 期细胞分布比例、细胞凋亡率、Ca2+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。

    Abstract:

    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 The Ca2+ 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 G1 phase cells, Bcl-2 mRNA and protein expression levels were decreased, and the distributions of S and G2 / 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, G1 phase cell distribution, snf Bcl-2 mRNA and protein expression levels were increased, whereas S and G2 / 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.

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王 衍,陈 玲,庞 胤,张海峰,李贺扬.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[J].中国比较医学杂志,2023,33(6):62~67,74.

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  • 收稿日期:2022-07-29
  • 在线发布日期: 2023-08-18
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