克拉霉素对糖氧剥离大鼠皮质神经细胞的保护作用及其抑制凋亡的机制研究

Study on the Protective Effect of Clarithromycin on Cortical Neurons Deprived of Oxygen and Glucose in Rats and Its Mechanism of Inhibition of Apoptosis

关键词:  克拉霉素  caspase-3  Bax  Bcl-2  糖氧剥离  皮质神经细胞

Key words:  clarithromycin  caspase-3  Bax  Bcl-2  oxygen glucose deprivation  cortical neuron

摘要:

目的 研究克拉霉素对大鼠皮质神经细胞糖氧剥离（oxygen glucose deprivation，OGD）后的保护作用及其与抑制凋亡相关的作用机制。方法 消化法提取原代大鼠皮质神经细胞，建立皮质神经细胞OGD模型，OGD前予以克拉霉素（clarithromycin，CAM）预处理；通过存活/凋亡检测试剂盒于荧光显微镜下观察各组细胞凋亡情况；试剂盒测定各组培养基LDH含量定量评估细胞凋亡情况；蛋白免疫印迹法测定各组细胞caspase-3，Bax，Bcl-2的表达情况。结果 OGD处理后皮质神经细胞凋亡及乳酸释放含量较正常组明显增加（P<0.01），100 μmol·L^-1 CAM预处理可显著减少OGD造成的细胞凋亡及LDH释放（P<0.01）；OGD处理后皮质神经细胞caspase-3，Bax/Bcl-2表达显著升高（P<0.01），而CAM预处理有效降低了caspase-3，Bax/Bcl-2表达。结论 克拉霉素对糖氧剥离大鼠皮质神经细胞具有保护作用，其作用可能与抑制了凋亡相关通路有关。

Abstract:

OBJECTIVE To research the protective effect of clarithromycin (CAM) on cortical neurons deprived of oxygen and glucose in rats and its related apoptosis mechanisms.METHODS Extraction of primary rat cortical neurons,to establish the oxygen glucose deprivation (OGD) model with CAM pretreatment;the survival/apoptosis detection kit was used to detect the cell apoptosis;LDH kit was used to measure the content of LDH in the medium of each group to evaluate the apoptosis.The expression of caspase-3,Bax and Bcl-2 in each group were detected by Western blot.RESULTS After treatment with OGD,compared with the control group,the apoptosis of cortical neurons and the content of lactic acid were significantly increased,the difference was significant (P<0.01);With 100 μmol·L^-1 CAM pretreatment the apoptosis and LDH release induced by OGD was significantly reduced (P<0.01).After treatment with OGD,the expression of caspase-3 and Bax/Bcl-2 in cortical neurons was significantly higher than that in control group (P<0.01),while CAM pretreatment effectively reduced the expression of caspase-3 and Bax/Bcl-2.CONCLUSION CAM has protective effect on cortical neurons deprived of oxygen and glucose,which may be related to the promotion of caspase-3,upregulation of Bax,downregulation of Bcl-2 protein on apoptosis related pathways.