摘要
目的探讨普罗布考对脑缺血再灌注(CI/R)大鼠脑损伤的减轻作用及对氧化应激的影响,阐明其可能作用机制。方法SD大鼠随机分为假手术组(生理盐水)、模型组(生理盐水)、普罗布考低(125 mg·kg^(-1)·d^(-1))、中(250 mg·kg^(-1)·d^(-1))、高剂量(500 mg·kg^(-1)·d^(-1))组,每组各15只,连续给药10 d后,除假手术组外,其余各组均采用线栓法构建大鼠动脉闭塞(MCAO)脑缺血模型,缺血2 h再灌注24 h后,利用Zea Longa法对各组大鼠进行神经功能评分,TTC染色法检测脑梗死体积,HE染色法观察缺血侧脑组织病理形态学变化,TUNEL染色法检测缺血侧脑皮质区细胞凋亡情况,比色分析法检测大鼠大脑皮质组织氧化应激因子丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)水平,Western Blot法检测脑组织中糖原合成酶激酶-3β(GSK3β)、磷酸化糖原合成酶激酶-3β(p-GSK3β)、转录因子NF-E2相关因子(Nrf-2)、血红素氧合酶(HO-1)、B细胞淋巴瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)的蛋白水平表达情况。结果与假手术组比较,模型组有明显的核仁固缩,形状改变,细胞排列散乱且有空泡,神经功能缺损评分、脑梗死体积及脑组织细胞凋亡率、氧化应激指标MDA水平、p-GSK3β和Bax蛋白表达显著升高,SOD活性及GSH含量、Nrf2、HO-1及Bcl-2表达水平显著降低(P<0.05);与模型组比较,普罗布考低、中、高剂量组均可显著降低脑神经功能缺损评分、脑梗死体积及细胞凋亡指数(P<0.05),对缺血侧脑组织的病理表现具有明显的改善作用,显著降低MDA含量,显著提高SOD、GSH水平,且均可显著促进Nrf2、HO-1及Bcl-2蛋白的表达,降低p-GSK3β活性及Bax蛋白的表达水平(P<0.05),另整体看来,普罗布考的作用效果具有一定的剂量依赖性。结论普罗布考可减轻大鼠CI/RI,促进抗凋亡因子的表达,并逆转SOD和GSH活性的下降、MDA含量的增加,其机制可能与抑制GSK3β磷酸化,激活Nrf2/HO-1通路,进而抑制氧化应激有关。
Objective To investigate the alleviating effect of probucol on brain injury and oxidative stress in cerebral ischemia reperfusion(CI/R)rats,and to clarify its possible mechanism.Methods SD rats were randomly divided into sham operation group(normal saline),model group(normal saline),probucol low(125 mg·kg^(-1)·d^(-1)),medium(250 mg·kg^(-1)·d^(-1))and high dose(500 mg·kg^(-1)·d^(-1))groups,with 15 rats in each group.After continuous administration for 10 days,except the sham operation group,the cerebral ischemia model of arterial occlusion(MCAO)was established by suture method in other groups,after ischemia for 2 hours and reperfusion for 24 hours,the neurological function of rats in each group was scored by Zea Longa method.The cerebral infarction volume was detected by TTC staining,the pathomorphological changes of cerebral tissue on the ischemic side were observed by HE staining,the apoptosis of cerebral cortex on the ischemic side was detected by TUNEL staining,the levels of oxidative stress factors malondialdehyde(MDA),superoxide dismutase(SOD)and reduced glutathione(GSH)in rat cerebral cortex were detected by colorimetric analysis,the protein levels of glycogen synthase kinase-3β(GSK3β),phosphorylated glycogen synthase kinase-3β(p-GSK3β),transcription related factor NF-E2(Nrf2),heme oxygenase(HO-1),B-cell lymphoma-2(Bcl-2)and Bcl-2 related X protein(Bax)were detected through the way of Western Blot.Results The model group showed obvious nucleolar pyknosis,shape change,scattered cell arrangement and vacuoles,neurological deficit score,cerebral infarction volume and apoptosis rate of brain tissue,oxidative stress index MDA level and p-GSK3β,the expression of SOD activity,GSH content,Nrf2,HO-1 and Bcl-2 decreased significantly(P<0.05)comparing with the sham-operated group.Compared with the model group,probucol low,medium and high dose group could significantly reduce the score of cerebral neurological deficit,cerebral infarction volume and apoptosis index(P<0.05),significantly improve the pathological manifestations of ischemic brain tissue,reduce the content of MDA and increase the levels of SOD and GSH,promote the expression of Nrf2,HO-1 and Bcl-2 proteins and reduce p-GSK3βactivity and Bax protein expression level significantly(P<0.05).On the whole,the effect of probucol was dose dependent.Conclusions Probucol can reduce CI/RI in rats,promote the expression of anti apoptotic factors,reverse the decrease of SOD and GSH activities and the increase of MDA content.Its mechanism may be related to the inhibition of GSK3βphosphorylation,activate Nrf2/HO-1 pathway,and then inhibit oxidative stress.
作者
张杰
王建平
赵洪云
ZHANG Jie;WANG Jianping;ZHAO Hongyun(Department of CT Diagnosis,Cangzhou Central Hospital,Cangzhou 061000,China)
出处
《临床神经外科杂志》
2023年第3期307-312,共6页
Journal of Clinical Neurosurgery
基金
河北省医学科学研究课题计划(20200305)。
