【摘要】 目的观察川芎抗脑缺血的神经胶质细胞Ube3a-as机制。方法取清洁级昆明种小鼠66只,随机分为川芎组(n=22)、模型组(n=22)、对照组(n=22)。川芎组、模型组采用线栓法建立昆明种小鼠大脑中动脉阻塞(MCAO)局灶性缺血再灌注模型。川芎组利用立体定位仪在侧脑室注射川芎提取物治疗,模型组不接受任何治疗。采用Bederson评分法对小鼠神经学进行评估,并分离培养海马神经胶质细胞,通过GFAP免疫化学分析鉴定原代神经胶质细胞的纯度,分析川芎抗脑缺血的神经胶质细胞Ube3a-as机制。结果 3组小鼠建模前Bederson评分比较差异无统计学意义(P>0.05)。川芎组与模型组建模后1天Bederson评分均显著高于对照组(P<0.05)。建模后1 d、2 d以及处死前,川芎组经治疗后小鼠Bederson评分明显低于模型组(P<0.05)。川芎组昆明小鼠梗死面积明显小于模型组(P<0.05),对照组昆明小鼠未建立模型,未见梗死面积。镜下可见原代小鼠海马星形胶质细胞生长良好,所有细胞均贴壁生长。免疫组化染色法鉴定结果显示,海马星形胶质细胞的GFAP免疫阳性反应率均在98.0%以上。PCR检测结果显示,川芎组小鼠海马组织中神经胶质细胞Ube3a-as水平显著高于模型组(川芎组:2.47±1.21;模型组:1.01±0.27,P<0.05)。结论脑缺血昆明种小鼠采用川芎提取物治疗效果良好,药物能直接影响神经胶质细胞Ube3a-as基因水平。更多还原

【Abstract】 Objective To observe the mechanism of chuanxiong against cerebral ischemic glial cells Ube3 a-as. Methods66 Kunming mice with the clean grade were selected and randomly divided into chuanxiong group(n=22), model group(n=22) and control group(n=22). The thread-occluded method was used to establish a model of middle cerebral artery occlusion(MCAO) focal cerebral ischemia and reperfusion of Kunming mice in chuanxiong group and the model group.Chuanxiong group was treated with injection of chuanxiong extract by stereotaxic device in the lateral ventricle. The model group did not receive any treatment. Mice neurology was assessed using the Bederson score method. The hippocampal glial cells were isolated and cultured. And the mechanism of chuanxiong against cerebral ischemic glial cells Ube3 a-as was analyzed through identifying the purity of primary glial cells with GFAP immunochemical analysis. Results There was no significant difference in the Bederson score among the three groups before modeling(P>0.05). The scores of Bederson in the chuanxiong group and model group at 1 day after modeling were significantly higher than those in the control group(P<0.05). The Bederson score of the mice in chuanxiong group after treatment was significantly lower than that in model group at 1 day, 2 days of modeling and before the execution(P<0.05). The infarct area of Kunming mice in chuanxiong group was significantly lower than that of the model group(P<0.05). The Kunming mice in the control group did not establish model. The infarct area did not occur in Kunming mice of the control group. It can be seen that the primary mice glioblast astrocytes were well grown and all cells were adherent under the microscope. Immunohistochemical staining identification results showed that the GFAP immune positive response rate of hippocampus astrocytes was above 98.0%. The level of Ube3 a-as in glial cells of hippocampus in chuanxiong group(2.47±1.21) was significantly higher than that of model group(1.01±0.27)(P<0.05). Conclusion The treatment effect of the extract of chuanxiong on Kunming mice with cerebral ischemia is good, and the drug can directly affect the Ube3 a-as gene level of glial cells.更多还原