龙琥醒脑颗粒介导MAPK通路对脑缺血再灌注损伤模型大鼠脑神经细胞的保护作用

Protective Effects of Longhu Xingnao Granules on Neural Cells in a Rat Model of Cerebral Ischemia Reperfusion Injury via the MAPK Pathway

目的:基于MAPK通路研究龙琥醒脑颗粒对脑缺血再灌注损伤(CIRI)模型大鼠脑神经细胞的保护作用。方法:采用大脑中动脉阻塞(MCAO)法制备CIRI模型,将造模成功的15只大鼠随机分为模型组、龙琥醒脑颗粒组和金纳多组,每组5只,另设置假手术组5只。龙琥醒脑颗粒组灌胃300 mg/kg龙琥醒脑颗粒,金纳多组灌胃150 mg/kg金纳多,假手术组和模型组灌胃等体积生理盐水,各组均干预5 d。评估各组大鼠神经功能评分,TTC染色法检测脑组织梗死面积,RT-PCR法检测各组大鼠脑组织中MAPK、p-ERK、p-CREB mRNA水平,Western blot检测各组大鼠脑组织中MAPK、p-ERK、p-CREB、Bcl-2、Bax和Casapase-3蛋白表达。结果:与假手术组比较,模型组大鼠神经功能损伤更加严重,神经功能评分、脑梗死面积、Bax和Casapase-3蛋白表达均显著升高(P<0.05),MAPK、p-ERK和p-CREB mRNA和MAPK、Bcl-2、p-ERK、p-CREB蛋白表达均下降(P<0.05)。与模型组相比,龙琥醒脑颗粒组和金纳多组大鼠神经功能评分、脑梗死面积、Bax和Casapase-3蛋白表达均显著下降(P<0.05),MAPK、pERK、p-CREB mRNA和MAPK、Bcl-2、p-ERK、p-CREB蛋白表达均显著上升(P<0.05),其中龙琥醒脑颗粒组最为显著。结论:龙琥醒脑颗粒可以改善CIRI模型大鼠的脑神经功能评分、降低脑梗死面积和细胞凋亡发挥对脑缺血再灌注损伤模型大鼠神经保护作用,其机制可能与调控MAPK通路有关。

Objective:To investigate the neuroprotective effects of Longhu Xingnao Granules on cerebral ischemia-reperfusion injury(CIRI)in rats based on the MAPK pathway.Methods:The CIRI model was established using the middle cerebral artery occlusion(MCAO)method.Fifteen successfully modeled rats were randomly divided into the model group,Longhu Xingnao Granules group,and Ginaton group,with 5 rats in each group.An additional sham-operated group(5 rats)was also included.The Longhu Xingnao Granules group was administered 300 mg/kg Longhu Xingnao Granules by gavage,the Ginaton group received 150 mg/kg Ginaton,and the sham-operated and model groups were given an equal volume of saline,all for 5 days.Neurological function scores were assessed,brain infarct volume was measured using TTC staining,and MAPK,p-ERK,and p-CREB mRNA levels in brain tissue were detected using RT-PCR.Protein expressions of MAPK,p-ERK,p-CREB,Bcl-2,Bax,and Caspase-3 were determined by Western blot.Results:Compared with the sham-operated group,the model group showed significantly worse neurological damage,with increased neurological function scores,brain infarct volume,and Bax and Caspase-3 protein expressions(P<0.05),while MAPK,p-ERK,and p-CREB mRNA levels and MAPK,Bcl-2,p-ERK,and p-CREB protein expressions were significantly decreased(P<0.05).Compared with the model group,the Longhu Xingnao Granules and Ginaton groups showed significant reductions in neurological function scores,brain infarct volume,and Bax and Caspase-3 protein expressions(P<0.05),and significant increases in MAPK,p-ERK,p-CREB mRNA levels and MAPK,Bcl-2,p-ERK,and p-CREB protein expressions(P<0.05),with the Longhu Xingnao Granules group showing the most pronounced effects.Conclusion:Longhu Xingnao Granules improve neurological function scores,reduce brain infarct volume,and decrease apoptosis in CIRI model rats,exerting neuroprotective effects likely via the regulation of the MAPK pathway.