| Objective Ischemic stroke is a clinical disease that causes limited brain tissue damage caused by blood supply disorders of the brain.Ischemic reperfusion injury refers to the rapid increase of tissue damage caused by massive oxygen free radicals generated by the ischemic cells after their blood flow recovers.It is the main pathogenesis of extensive tissue and cell damage.Currently it is believed that ischemic stroke is commonly observed in atherosclerosis(AS).Plaque shedding blocks cerebral blood vessels,thus causing a series of pathological reactions such as energy metabolism disturbance,oxidative stress injury,inflammatory response and apoptosis.Angong Niuhuang Pill is an emergency drug used in the treatment of ischemic stroke,but its mechanism of action has not been fully clarified.This study is to study the effect of Angong Niuhuang Pill on ischemic stroke from the perspective of the compatibility of Angong Niuhuang Pill and Middle cerebral artery occlusion(MCAO)reperfusion injury model.MethodsThrough TCMSP,BATMAN-TCM,Herb database and literature retrieval,the active constituents of each drug in Angong Niuhuang Pill were collected,and the target prediction was made in Swiss Target Prediction database.Relevant targets for ischemic stroke were collected through Drug Bank,Dis Ge NET,Gene Cards,and OMIM databases.The collected ischemic apoplexy-related targets were mapped to the targets of the active components of Angongniuhuang Pills to obtain potential targets of Angongniuhuang pills in the treatment of ischemic apoplexy.Then,the core targets of Angongniuhuang pills in the treatment of ischemic apoplexy were analyzed through Metascape online data.According to the analysis results of network pharmacology,animal experiments were designed to verify the results.The model of ischemia-reperfusion injury rats was established in 6-8 week-old SD male rats,which were divided into sham operation group,model group,Angong Niuhuang Wan group,Huophlegm and Qingrejiedu group.The nerve function of rats was observed by m NSS score;the infarct size of rats was observed by TTC staining;the brain damage of rats was observed by HE and Nizi staining;the apoptosis of rats was observed by TUNEL staining;the SOD and MDA contents of brain tissue were detected by kit to observe the oxidative stress level;the inflammation expression of brain tissue sections was observed under the microscope.Results A total of 150 effective components of Angong Niuhuang pills were collected,including 930 targets and 1171 targets for ischemic stroke.The target of Angong Niuhuang pills was mapped to the target of ischemic stroke,and 156 potential targets were obtained.The 156 targets were divided into the phlegm-clearing group and the heat-clearing and detoxification group for analysis.We could see that the pathways involved in the core targets included both common pathways and separate pathways in each group,and the pathways of the two groups of core targets were mainly related to inflammation,metabolism and nervous system regulation.Among them,the phlegm-clearing group focused more on the regulation of nervous system.In the experimental verification,we found that Angong Niuhuang Wan group,Huotan Kaiqiao group and Qingrejiedu group can all reduce the neurological function score of rats with ischemia reperfusion injury,reduce the infarct area of brain tissue after cerebral ischemia reperfusion injury,improve the brain tissue injury of rats with ischemia reperfusion injury,reduce the inflammation,oxidative stress level and apoptosis of brain tissue.In all the experimental results,the efficacy of Angong Niuhuang Pill group was better than that of Huophlegm Huophlegm opening orifice group and clearing heat and detoxifying group,while Huophlegm opening orifice group was slightly better than clearing heat and detoxifying group.ConclusionThe network pharmacological results showed that the core targets of Huophlegm Huophlegm Opening-body group and Qingrejiedu group were mainly involved in the regulation of inflammation,nervous system and substance metabolism,indicating that Angongniuhuang Pill may play the comprehensive effects of clearing heat and detoxifying,and clearing phlegm and opening-body by reducing inflammation,regulating nervous system and intervening energy metabolism.Moreover,in animal experiments,Angong Niuhuang Wan group,Huophlegm opening orifice group and Qingrejiedu group can all play a therapeutic role in protecting brain tissue nerves by improving brain nerve injury,reducing inflammation,oxidative stress level and cell apoptosis. |
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