摘要
目的:研究侧脑室注射Nogo-A反义寡核苷酸对大鼠脑缺血再灌注后蛋白激酶C(PKC)-Rho A信号通路的影响。方法:40只大鼠,随机分为假手术组(A组)、再灌注6 h组(B组)、再灌注24 h组(C组)、再灌注72 h组(D组)、再灌注1周组(E组)、再灌注2周组(F组)、Nogo-A 72 h组(G组)、Nogo-A 1周组(H组),每组5只。制备大鼠脑缺血再灌注模型,A组不造成缺血,B、C、D、E、F组缺血后分别再灌注6 h、24 h、72 h、1周、2周,G、H组分别在缺血后脑室注射Nogo-A反义寡核苷酸后再灌注72 h、1周。分别用RT-PCR及Western blot检测缺血周围区大脑皮质PKCγ、Rho A的m RNA及蛋白表达的变化。结果:与A组比较,B组大脑皮质PKCγm RNA表达明显增加(P<0.01),C组降至正常(P>0.05),D组出现第2次升高(P<0.01),E组最高(P<0.01),H组较E组表达明显下降(P<0.05)。与A组比较,B、D、E组Rho A m RNA表达明显增加(P<0.01),G、H组较D、E组表达明显下降(P<0.05)。与A组比较,B、D组的PKCγ蛋白表达增加(P<0.05),E组明显增加(P<0.01),G、H组较D、E组表达明显下降(P<0.05)。与A组比较,B、D、E组的Rho A蛋白表达增加,G、H组较D、E组表达明显下降(P<0.05)。结论:PKCγ及Rho A基因表达在大鼠脑缺血再灌注后有两个高峰,后一个高峰与Nogo A的升高相一致。脑室注射Nogo-A反义寡核苷酸能抑制PKCγ及Rho A的表达,表明缺血后Nogo-A的升高参与PKC/Rho A信号通路的激活。
Objective：To investigate the effects of Nogo-A antisence oligodeoxynucleotides on signal pathway of PKC-RhoA after cerebral ischemia-reperfusion in rats. Methods： Forty rats were randomly assigned to groups of sham-operated （A）, reperfusion 6 h （B）, reperfusion 24 h （C）, reperfusion 72 h （D）, reperfusion 1 week （E）, reperfusion 2 weeks （F）, Nogo-A 72 h （G） and Nogo-A 1 week （H）, with 5 rats in each group. The middle cerebral artery occlusion （MCAO） models was established. The group A was not induced ischemia, and the groups B, C, D, E, F were reperfused with 6 h, 24 h, 72 h, 1 week, 2 weeks respectively after ischemia. The groups G, H were given intracerebroventricularly with Nogo-A antisence oligodeoxynucleotides after the ischemia, then reperfused with 72 h or 1 week respectively. RT-PCR was used to determine the changes of PKCγand RhoA mRNA expression while the Western blot technique was used to detect the PKCγand RhoA protein expression in the periphery of infarction cerebral cortex. Results： The expression level of PKCγ mRNA in group B were obviously increased compared with group A （ 〈0.01） and group C returned to normal （ 〉0.05）. The expression of PKCγ mRNA elevated again in group D （ 〈0.01）. It was highest in group E（ 〈0.01）. The expression of PKCγmRNA in group H were significantly declined compared with group E （P〈0.05）. The expressions of RhoA mRNA in group B, D,were higher than those in group A （ 〈0.01）. In group G and H, it was significantly declined compared with group D and E（ 〈0.05）. The expression of PKCγprotein were higher in group B,D than that in group A（ 〈0.05）. The increase was significant in group E （ 〈0.01）. The expression of PKCγprotein in group G and H were significantly declined in comparison with group D and E（ 〈0.05）. The expression of RhoA protein were increased in group B, D, E com-pared with that of group A （ 〈0.05）, which was significantly declined tin group G and H compared with group D and E （ 〈0.05）. Conclusion：The expression of PKCγand RhoA gene showed two peaks and the second increase was coincident with the change of Nogo-A after cerebral ischemia-reperfusion. Intracerebroventricular injection of Nogo-A antisence oligodeoxynucleotides can inhibit this increase,suggesting that Nogo-A is involved in the activa-tion of PKC/RhoA signal pathway.
出处
《神经损伤与功能重建》
2014年第6期468-471,共4页
Neural Injury and Functional Reconstruction
