抗Nogo-A抗体对急性脑梗死神经轴突再生影响的研究

发布时间：2019-06-04 18:52

【摘要】：目的：探讨抗Nogo-A抗体对大鼠实验性大脑中动脉阻塞(MCAO)后脑梗死的治疗效果,动态观察脑梗死后Nogo-A蛋白表达的变化。方法：180只SD大鼠随机分为假手术组、IgG组、mNGF组、Nogo-A抗体组,每组45只。线栓法建立大鼠MCAO模型,侧脑室置管给药(8ul/次,1次/同)治疗,其中前两组均给予正常大鼠IgG治疗,mNGF组给予鼠神经生长因子(mNGF)治疗,Nogo-A抗体组予抗Nogo-A抗体治疗。各组分三个亚组(每亚组15只),分别存活1d、3d、7d,并于各时间点进行神经功能评分,随后断头取脑。脑标本分别行脑梗死相对体积测定,Nogo-A、GAP-43、NF200蛋白免疫荧光检测,Nogo-A、GAP-43、NF200蛋白免疫印迹检测与Nogo-A mRNA、GAP-43mRNA、NF200mRNA实时荧光定量PCR检测。结果：假手术组各时间点神经功能评分正常,其余三组在治疗后1d、3d两个时间点神经功能评分组间比较差异无统计学意义(P0.05),但mNGF组、Nogo-A抗体组治疗7d后神经功能评分均较IgG组明显降低(p0.05),其中Nogo-A抗体组最明显。假手术组没有脑梗死,其余三组治疗后各个时间点脑梗死相对体积组间比较差异无统计学意义(P0.05)。1d、3d和7d时间点,假手术组Nogo-A、NF-200、GAP-43蛋白表达差异无统计学意义(p0.05);1d和3d时间点,IgG组、mNGF组和Nogo-A抗体组Nogo-A、NF-200、GAP-43蛋白表达组间比较差异无统计学意义(p0.05);7d时间点,Nogo-A抗体组分别与IgG、mNGF组比较Nogo-A蛋白表达明显减低(p0.05),Nogo-A抗体组、mNGF组分别与IgG组比较NF-200、GAP-43蛋白表达明显升高(p0.05),但Nogo-A抗体组与mNGF组比较NF-200、GAP-43蛋白表达差异无统计学意义(p0.05)。1d、3d和7d时间点,假手术组Nogo-A、NF-200、GAP-43mRNA表达差异无统计学意义(p0.05);1d时间点,IgG组、mNGF组和Nogo-A抗体组组间比较Nogo-A mRNA表达差异无统计学意义(p0.05);3d和7d时间点,Nogo-A抗体组分别与IgG、 mNGF组比较Nogo-A蛋白表达明显减低(p0.05),Nogo-A抗体组、mNGF组分别与IgG组比较NF-200、GAP-43mRNA表达明显升高(p0.05),但Nogo-A抗体组与mNGF组比较NF-200、GAP-43蛋白表达差异无统计学意义(p0.05)。结论：应用抗Nogo-A抗体治疗,通过促进轴突再生改善神经缺损功能的同时并不相应减少脑梗死的体积。由于脑梗死后7d Nogo-A蛋白仍然高表达,所以抗Nogo-A蛋白抗体治疗至少要持续7天以上。
[Abstract]:Aim: to investigate the therapeutic effect of anti-Nogo-A antibody on cerebral infarction after experimental middle cerebral artery occlusion (MCAO) in rats, and to observe the changes of Nogo-A protein expression after cerebral infarction. Methods: 180 SD rats were randomly divided into sham operation group, IgG group, mNGF group and Nogo-A antibody group with 45 rats in each group. The rat model of MCAO was established by thread thrombus method, and the lateral ventricle catheterization (8ul/ times, once / the same) was used to treat the rats. The first two groups were treated with IgG, and the mNGF group was treated with nerve growth factor (mNGF). Nogo-A antibody group was treated with anti-Nogo-A antibody. Each group was divided into three subgroups (15 rats in each subgroup), survived for 1 day, 3 days and 7 days respectively, and scored the neurological function at each time point, and then decapitated the brain. The relative volume of cerebral infarction, Nogo-A,GAP-43,NF200 protein immunofluorescence, Nogo-A,GAP-43,NF200 protein immunoblotting and Nogo-A mRNA,GAP-43mRNA,NF200mRNA real-time fluorescence quantitative PCR were measured respectively. Results: the neurological function scores of the false operation group were normal at each time point, and there was no significant difference in the neurological function scores of the other three groups at 3 days after treatment (P 0.05), but there was no significant difference in the neurological function scores of the other three groups at 3 days after treatment (P 0.05), but in the mNGF group, After 7 days of treatment, the neurological function scores of Nogo-A antibody group were significantly lower than those of IgG group (p0.05), especially in Nogo-A antibody group. There was no cerebral infarction in the pseudo-operation group, but there was no significant difference in the relative volume of cerebral infarction in the other three groups at each time point after treatment (P 0.05). At 1 d, 3 d and 7 d, Nogo-A,NF-200, in the pseudo-operation group was not significantly different from that in the other three groups at each time point after treatment. There was no significant difference in the expression of GAP-43 protein (p0.05). There was no significant difference in the expression of Nogo-A,NF-200,GAP-43 protein between IgG group, mNGF group and Nogo-A antibody group at 1 d and 3 d (p0.05). At 7 d, the expression of Nogo-A protein in Nogo-A antibody group was significantly lower than that in IgG,mNGF group (p0.05), while the expression of NF-200,GAP-43 protein in Nogo-A antibody group and mNGF group was significantly higher than that in IgG group (p0.05). However, there was no significant difference in the expression of NF-200,GAP-43 protein between Nogo-A antibody group and mNGF group (p0.05). There was no significant difference in Nogo-A,NF-200,GAP-43mRNA expression in sham operation group at 1 d, 3 d and 7 d (p0.05). At 1 d, there was no significant difference in the expression of Nogo-A mRNA between IgG group, mNGF group and Nogo-A antibody group (p0.05). At 3 d and 7 d, the expression of Nogo-A protein in Nogo-A antibody group was significantly lower than that in IgG, mNGF group (p0.05), while the expression of NF-200,GAP-43mRNA in Nogo-A antibody group and mNGF group was significantly higher than that in IgG group (p0.05). However, there was no significant difference in the expression of NF-200,GAP-43 protein between Nogo-A antibody group and mNGF group (p0.05). Conclusion: anti-Nogo-A antibody can improve the function of nerve defect by promoting axonal regeneration and does not reduce the volume of cerebral infarction at the same time. Because the expression of Nogo-A protein is still high on the 7th day after cerebral infarction, anti-Nogo-A protein antibody therapy should last for at least 7 days.
【学位授予单位】：贵阳中医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R743.33

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