奥拉西坦通过Wnt/β-catenin减轻大鼠脑缺血再灌注损伤的实验研究

Oxiracetam attenuates cerebral ischemia-reperfusion injury in rats by Wnt/β-catenin

目的研究奥拉西坦对大鼠脑缺血再灌注(I/R)损伤的影响及分子机制。方法 2017年5月-2018年12月在上海市第六人民医院南院中心实验室进行实验,选择成年雄性SD大鼠并随机分为假手术组(n=10)、I/R组(n=10)、奥拉西坦组(n=10),后2组采用线栓法建立脑I/R模型,奥拉西坦组给予奥拉西坦200 mg/kg腹腔注射,连续7 d。比较3组间神经功能缺损评分、血清神经元特异性烯醇化酶(NSE)及星形胶质原蛋白(S100B)含量、脑组织中细胞凋亡率及凋亡基因、Wnt/β-catenin通路基因表达的差异。结果 I/R组大鼠的神经功能缺损评分、血清NSE及S100B含量、脑组织的细胞凋亡率及Bcl-2相关X蛋白(Bax)、细胞色素C(CytC)、含半胱氨酸的天冬氨酸蛋白水解酶-3(Caspase-3)、糖原合成酶激酶-3β(GSK-3β)的表达水平均明显高于假手术组,脑组织中B淋巴细胞瘤-2基因(Bcl-2)、Wnt1、Wnt3、β-连环蛋白(β-catenin)的表达水平均明显低于假手术组(P<0.05或0.01);奥拉西坦组大鼠的神经功能缺损评分、血清NSE及S100B含量、脑组织的细胞凋亡率及Bax、CytC、Caspase-3、GSK-3β的表达水平均明显低于I/R组,脑组织中Bcl-2、Wnt1、Wnt3、β-catenin的表达水平均明显高于I/R组(P<0.05或0.01)。结论奥拉西坦能够减轻大鼠脑I/R损伤的程度,且该作用与Wnt/β-catenin通路的激活有关。

Objective To study the effects of Oxiracetam on cerebral ischemia reperfusion(I/R)injury in rats and its molecular mechanism.Methods From May 2017 to December 2018,the experiment was carried out in the Central Laboratory of Southern Hospital of Shanghai Sixth People’s Hospital.Adult male SD rats were randomly divided into sham-operated group(n = 10),I/R group(n= 10)and olacetam group(n = 10).Brain I/R model was established by thread embolization in the latter two groups,and olacetam group was given olacetam 200 mg/kg intraperitoneal injection.Shoot for 7 days.The differences of neurological deficit score,serum neuron specific enolase(NSE)and astroglial protein(S100 B),apoptotic rate of brain tissue,apoptotic gene and Wnt/β-catenin pathway gene expression were compared among the three groups.Results Neurological deficit score,serum NSE and S100 B content,brain tissue apoptosis rate and Bcl-2 related X protein(Bax),cytochrome C(CytC),cysteine-containing aspartate in I/R group The expression levels of Caspase-3 and glycogen synthase kinase-3β(GSK-3β)were significantly higher than those in the sham operation group.The expression levels of B lymphocyte 2 gene(Bcl-2),Wnt1,Wnt3,and β-catenin in brain tissue were significantly lower than those in the sham operation group.The neurological deficit score,serum NSE and S100 B content,brain tissue apoptosis rate and expression levels of Bax,CytC,Caspase-3 and GSK-3β in oxiracetam group were significantly lower than those in I/R group(P<0.01).The expression levels of Bcl-2,Wnt1,Wnt3 and β-catenin in brain tissue were significantly higher than those in I/R group(P<0.01).Conclusion Oxiracetam can reduce the degree of brain I/R injury in rats,and this effect is related to the activation of Wnt/beta catenin pathway.