目的：探讨外源性一氧化碳释放剂（CORM-2）诱导大鼠神经细胞凋亡及醒脑静注射液对其干预作用的机制。方法：在光学显微镜下观察CORM-2 对神经细胞形态的影响；通过MTT 法检测CORM-2对细胞存活率的影响；应用流式细胞仪检测细胞凋亡率；以Western Blotting 检测相关蛋白的表达。同时观察醒脑静注射液对神经细胞形态，细胞存活率及相关蛋白表达的影响。结果：CORM-2 影响神经细胞存活率呈时间依赖性和剂量依赖性；100、200、400、800 μmol·L-1 CORM-2 作用于神经细胞24 h 后，可使细胞存活率逐渐降低；可以诱导神经细胞凋亡及Caspase 依赖性线粒体途经相关蛋白Caspase-3、Caspase-9 及Cytochrome C（Cyt C）的活化，且呈剂量依赖性。选择200 μmol·L-1 CORM-2 对皮层神经细胞进行诱导损伤，经醒脑静注射液孵育神经细胞20 h 后，10 mL·L-1 及20 mL·L-1 醒脑静注射液可明显降低细胞早期凋亡率，细胞Caspase-3、Caspase-9 与Cyt C 的表达逐渐降低。结论：CORM-2 可能通过激活Caspase 依赖性线粒体途径诱导神经细胞凋亡，醒脑静注射液可抑制细胞凋亡的线粒体途径从而干预CORM-2 诱导的神经细胞凋亡。

This study was aimed to investigate the mechanism of CO -releasing molecules (CORM -2) induced apoptosis and the intervention effect of Xingnaojing (XNJ) Injection in neuronal cells of rats. Optical microscope was applied to observe morphologic changes of neuronal cells. MTT assay was performed to assess the survival rates of CORM-2 on neuronal cells. Apoptosis was examined by flow cytometric analysis and the expression of relative proteins was measured by western blotting analysis. At the same time the morphologic changes, survival rates and expression of relative proteins of neuronal cells were also checked after XNJ treatment. The results showed that CORM-2 can influence survival rates in a time- and concentration-dependent manner. Survival rates decreased gradually after the cultures subjected to 24 h with 100 μmol·L-1, 200 μmol·L-1, 400 μmol·L-1 and 800 μmol·L-1 of CORM-2. It can induce neuronal apoptosis and activate Caspase-3, Caspase-9 and Cytochrome C in a concentration-dependent manner. The neuronal cells were treated with 200 μmol·L-1 of CORM-2 and then incubated with 10 mL·L-1 and 20 mL·L-1 XNJ injection for 20 h. It turned out early neuronal apoptosis decreased and the expression of Caspase-3, Caspase-9 and Cytochrome C also decreased. To sum up, CORM-2 may induce neuronal apoptosis through Caspase-dependent mitochondrial pathway, which can be intervened by XNJ Injection through inhibiting Caspase-dependent mitochondrial pathway.

北京中医药大学自主选题项目（2009JYBZZ-JS041）：醒脑静注射液对一氧化碳暴露诱导的神经细胞凋亡的干预作用及机制分析，负责人：王晶。