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目的观察脑出血后氧化应激与神经细胞凋亡的相关性及对凋亡相关蛋白c-myc表达的影响。方法选择SD雄性大鼠84只,随机分为假手术组、模型组、MPEG-SOD组,每组28只,自体血注入法建立脑出血模型,MPEG-SOD组建模前腹腔注射MPEG-SOD;术后12 h、1 d、3 d、7 d不同时间点行神经功能缺陷评分(NDS),取脑组织测定脑组织含水量,分光光度法测定丙二醛(MDA)、超氧化物歧化酶(SOD),免疫组化法测定核因子κB(NF-κB)p65阳性细胞表达,TUNEL测定神经细胞凋亡,Western blot测定c-myc蛋白表达水平。结果与假手术组比较,模型组各时点NDS评分、SOD活性明显降低,脑组织含水量、MDA含量、c-myc蛋白、NF-κB p65、TUNEL阳性细胞表达明显升高(P<0.05),术后3 d达峰值,与模型组同时点比较,MPEG-SOD组NDS评分、SOD活性明显升高,脑组织含水量、MDA含量、c-myc蛋白、NF-κB p65、TUNEL阳性细胞表达明显降低(P<0.05),cmyc蛋白表达与TUNEL阳性细胞数呈正相关(模型组r=0.83,MPEG-SOD组r=0.91,P<0.05)。结论脑出血后氧化应激诱导神经细胞凋亡可能与启动凋亡相关基因c-myc有关。
Objective To observe the correlation between oxidative stress and neuronal apoptosis after intracerebral hemorrhage and its effect on the expression of apoptosis related protein c-myc. Methods Eighty-four male Sprague-Dawley rats were randomly divided into sham operation group, model group and MPEG-SOD group, 28 rats in each group. The model of intracerebral hemorrhage was established by autologous blood injection. MPEG-SOD The neurological deficit score (NDS) at 12 h, 1 d, 3 d, and 7 d after operation were measured. Brain water content was measured. The content of malondialdehyde (MDA), superoxide dismutase The expression of nuclear factor-kappaB (NF-κB) p65 positive cells was detected by immunohistochemistry. Neuronal apoptosis was measured by TUNEL. The expression of c-myc protein was detected by Western blot. Results Compared with the sham operation group, the NDS score and the SOD activity in model group decreased significantly at each time point. The content of water content, MDA content, c-myc protein, NF-κB p65 and TUNEL positive cells in the model group were significantly increased (P <0.05) , Peaked on the 3rd postoperative day. Compared with the model group, NDS score and SOD activity in MPEG-SOD group were significantly increased. The content of water content, MDA content, c-myc protein, NF-κB p65, TUNEL positive cells (P <0.05). The expression of cmyc protein was positively correlated with the number of TUNEL positive cells (r = 0.83 in model group and r = 0.91 in MPEG-SOD group, P <0.05). Conclusion Oxidative stress-induced neuronal apoptosis after intracerebral hemorrhage may be related to the initiation of apoptosis-related gene c-myc.