依达拉奉右莰醇通过PKC/ERK通路对脑缺血再灌注大鼠发挥脑保护作用  

Edaravone dexborneol exerts cerebral protective effect on cerebral ischemia-reperfusion rats through PKC/ERK pathway

作　　者：张静[1] 钱倩 白艳梅 ZHANG Jing;QIAN Qian;BAI Yanmei(Fifth Department of Neurology,Baoding First Central Hospital,Baoding,Hebei 071000,China)

机构地区：[1]保定市第一中心医院神经内五科,河北省保定市071000

出　　处：《中国动脉硬化杂志》2023年第5期391-398,共8页Chinese Journal of Arteriosclerosis

基　　金：河北省医学科学研究重点课题计划项目(20180003);河北省卫生健康委医学科学研究课题(20220291)。

摘　　要：[目的]观察依达拉奉右莰醇(EDDE)对脑缺血再灌注大鼠神经元凋亡的影响,并探讨其作用机制。[方法]将SD大鼠随机分为假手术组、大脑中动脉闭塞(MCAO)组、低剂量EDDE组(EDDE-L组,3 mg/kg)、高剂量EDDE组(EDDE-H组,6 mg/kg)、白屈菜红碱(CHE,PKC抑制剂)组(5 mg/kg CHE)、EDDE+CHE组(6 mg/kg EDDE+5 mg/kg CHE),每组15只。除假手术组外,其余各组大鼠采用线栓法构建MCAO模型。对各组大鼠进行神经功能缺损评分;再灌注24 h后,TTC染色检测大鼠脑梗死体积;HE染色观察皮质神经细胞病理损伤;TUNEL染色检测皮质神经细胞凋亡;免疫组织化学法检测皮质区神经细胞Bcl-2、Bax的表达;Western blot检测脑组织Caspase-3、cleaved Caspase-3和蛋白激酶C(PKC)/细胞外信号调节激酶(ERK)通路相关蛋白(PKC、p-PKC、ERK1/2、p-ERK1/2)的表达。[结果]与假手术组相比,MCAO组大鼠神经功能缺损评分、脑梗死体积百分比和神经细胞凋亡率升高,Bax阳性表达增加,cleaved Caspase-3/Caspase-3比值升高(均P<0.05),Bcl-2阳性表达和Bcl-2/Bax比值降低,脑组织p-PKC/PKC、p-ERK1/2/ERK1/2比值降低(均P<0.05),脑皮质细胞排列稀疏,神经细胞肿胀、空泡样变性明显、核固缩,组织坏死;与MCAO组相比,EDDE-L组和EDDE-H组大鼠神经功能缺损评分、脑梗死体积百分比和神经细胞凋亡率降低,Bax阳性表达减少,cleaved Caspase-3/Caspase-3比值降低(均P<0.05),Bcl-2阳性表达升高,Bcl-2/Bax、p-PKC/PKC和p-ERK1/2/ERK1/2比值升高(均P<0.05),脑皮质病理损伤有不同程度的改善,神经细胞数量增多,空泡样变性减轻,胞核较清晰,且EDDE-H组的改善优于EDDE-L组;CHE可以消除EDDE的神经保护作用。[结论]EDDE可抑制神经细胞凋亡,减轻脑缺血再灌注损伤,其机制可能与激活PKC/ERK通路有关。Aim To observe the effect of edaravone dexborneol(EDDE)on neuronal apoptosis in rats with cerebral ischemia-reperfusion,and to explore its mechanism.Methods SD rats were randomly divided into sham group,middle cerebral artery occlusion(MCAO)group,low-dose EDDE group(EDDE-L group,3 mg/kg),and high-dose EDDE group(EDDE-H group,6 mg/kg),chelerythrine(CHE,PKC inhibitor)group(5 mg/kg CHE),EDDE+CHE group(6 mg/kg EDDE+5 mg/kg CHE),with 15 rats in each group.Except for the sham group,the rats in the other groups were given the suture method to construct the MCAO model.The rats in each group were scored for neurological deficits;after 24 hours of reperfusion,TTC staining was used to detect the cerebral infarct volume of rats;HE staining was used to observe the pathological damage of cortical nerve cells;TUNEL staining was used to detect cortical nerve cells apoptosis;immunohistochemical method was used to detect the expression of Bcl-2 and Bax in the cortex nerve cells;Western blot was used to detect the expression of Caspase-3,cleaved Caspase-3 and protein kinase C(PKC)/extracellular signal-regulated kinase(ERK)pathway related proteins(PKC,p-PKC,ERK1/2 and p-ERK1/2)in brain tissue.Results Compared with sham group,the neurological deficit score,the percentage of cerebral infarction volume and the apoptosis rate of nerve cells increased,the positive expression of Bax increased and the ratio of cleaved Caspase-3/Caspase-3 increased(all P<0.05),the positive expression of Bcl-2 decreased,the ratio of Bcl-2/Bax,p-PKC/PKC and p-ERK1/2/ERK1/2 decreased in brain tissue(all P<0.05),the cerebral cortex cells were sparsely arranged,swelling and vacuolar degeneration of nerve cells were obvious,the nucleus shrank and the tissue becomes necrotic in MCAO group.Compared with MCAO group,the neurological deficit score,cerebral infarction volume percentage and apoptosis rate of nerve cells decreased,the positive expression of Bax decreased,the ratio of cleaved Caspase-3/Caspase-3 decreased(all P<0.05),the positive expression of Bcl-2 incre

关 键 词：依达拉奉右莰醇 脑缺血再灌注损伤 神经细胞 蛋白激酶C 细胞外信号调节激酶 

分 类 号：R741[医药卫生—神经病学与精神病学] R5[医药卫生—临床医学]