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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),100μmol·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。
Objective To study the protective effect of clarithromycin on oxygen-glucose deprivation (OGD) in rat cortical neurons and its mechanism of action related to the inhibition of apoptosis. Methods Primary rat cortical neurons were isolated by digestion method. OGD model of cortical neurons was established and pretreated with clarithromycin (CAM) before OGD. The apoptosis of cells in each group was observed by fluorescence microscope The LDH levels in each group were assayed by kit to evaluate the cell apoptosis quantitatively. The expression of caspase-3, Bax and Bcl-2 in each group were determined by Western blotting. Results The levels of apoptosis and lactic acid release in cortical neurons after OGD treatment were significantly higher than those in normal group (P <0.01). Pretreatment with 100 μmol·L -1 CAM significantly reduced the apoptosis and LDH release induced by OGD (P < 0.01). The expressions of caspase-3 and Bax / Bcl-2 were significantly increased in OGD-treated cortical neurons (P <0.01), while CAM pretreatment reduced the expressions of caspase-3 and Bax / Bcl-2. Conclusion Clarithromycin can protect the cortical neurons from glucose-stripping rats, which may be related to the inhibition of apoptosis-related pathways.