The Efficacy And Neuroprotective Mechanism Of Guizhi Fuling Pill In The Treatment Of Acute Cerebral Infarctio

Cerebral infarction(cerebral infarction),also known as ischemic stroke,is a kind of clinical syndrome in which various cerebrovascular diseases lead to blood supply disorders in the brain,ischemic and hypoxic necrosis of local brain tissue,and rapid occurrence of corresponding neurological deficits.Acute cerebral infarction(ACI)is the most common type of stroke.Although most patients can receive timely treatment,inpatients still have high mortality and disability rates.Intravenous thrombolysis,arterial thrombolysis and mechanical thrombectomy are the most commonly used and effective treatment methods in clinical practice.However,due to the limitation of its treatment time window and contraindications,fewer patients are eligible to receive corresponding treatment.After exceeding the treatment time window,the treatment method is limited,and other treatment methods to improve cerebral blood circulation also have a certain degree of bleeding risk.Therefore,it is very important to find a more effective treatment.TCM treatment of ACI is widely used in clinical practice,and there is no time window limit.ACI belongs to ’stroke’ in traditional Chinese medicine.Modern Chinese medicine research shows that the main pathogenesis of stroke is’wind’,’fire’,’phlegm’,’blood stasis’ and ’deficiency’.Under certain conditions,they interact with each other and eventually lead to ’blood stasis’.Among them,’stasis’ is considered to be the most critical pathological factor throughout the whole course of ’stroke’.The method of promoting blood circulation and removing blood stasis can effectively improve the prognosis of ACI patients,protect nerve function,promote nerve function recovery,and reduce disability rate.Guizhi Fuling Wan was first recorded in Zhang Zhongjing’s ’Synopsis of the Golden Chamber’.It is mainly used to treat gynecological diseases with syndrome of static blood blocking in uterus,and the main pathological product of stroke is also blood stasis.Although the disease location is different,the treatment mechanism is similar.Guizhi Fuling Wan has the effects of reducing blood viscosity,anti-inflammation,immune regulation,protecting vascular endothelial cells,anti-oxidation and improving circulation.A number of studies have shown that Guizhi Fuling Wan has significant clinical efficacy in the treatment of ACI.However,in recent years,the research on Guizhi Fuling Wan in the treatment of ACI is mainly based on clinical experiments,and there is little research on its mechanism.Therefore,this study summarizes the efficacy of Guizhi Fuling Wan in the treatment of ACI from multiple perspectives by means of systematic review and meta-analysis,and explores its therapeutic mechanism through network pharmacology and animal experiments,so as to provide theoretical basis for its application in the clinical treatment of ACI in traditional Chinese medicine.Clinical Research:Systematic Review and Meta-analysis of Guizhi Fuling Wan in the Treatment of Acute Cerebral InfarctionObjective:To summarize and explore the curative effect of Guizhi Fuling Wan in the treatment of ACI.Methods:In clinical research,we used systematic evaluation and meta-analysis to screen and analyze the published clinical literature on Guizhi Fuling Wans in the treatment of ACI.Randomized controlled clinical trials of Guizhi Fuling Wan in the treatment of ACI were searched in Wanfang,CNKI,CBM and Pubmed databases up to January 1,2023.According to the inclusion and exclusion criteria,the eligible literature was selected for meta-analysis,and the risk of bias of the literature was evaluated.Result:A total of 16 studies were included.The results of meta-analysis showed that the total effective rate of the treatment group[RR=1.20,95%CI(1.13,1.27),Z=3.44,P<0.00001],NIHSS score[MD=-2.47,95%CI(-3.33,-1.62),Z=5.67,P<0.00001],quality of life score[MD=10.92,95%CI(7.41,14.43),Z=6.10,P<0.00001].The mRS score[MD=-0.59,95%CI(-0.90,-0.27),Z=3.64,P=0.0003],TNF-α content[MD=-0.48,95%CI(-0.50,-0.45),Z=36.72,P=<0.00001],IL-6 content[MD=-15.51,95%CI(-25.72,-5.30),Z=2.98,P=0.003]were better than the control group.Conclusion:The results showed that Guizhi Fuling Wan could improve the clinical efficacy,reduce neurological deficits,improve activities of daily living,and reduce the content of related serum inflammatory factors in the treatment of acute cerebral infarction.Experiment 1:To explore the mechanism of Guizhi Fuling Wan in the treatment of acute cerebral infarction based on network pharmacologyObjective:To explore the mechanism of Guizhi Fuling Wan in the treatment of acute cerebral infarction based on the research method of network pharmacology.Methods:The main active components of Guizhi Fuling Wans were collected in tcmsp database.The screening conditions were oral bioavailability(OB)≥30%and drug-likeness(DL)≥0.18.Using genecards,OMIM and drugbank databases to collect the action targets of acute cerebral infarction,obtain the intersection targets of Guizhi Fuling Wan and disease,and draw the intersection Wayne diagram.The PPI network was constructed by bisogenet,and the core targets were extracted by Cytoscape.After obtaining key target proteins,GO function enrichment analysis and KEGG analysis were analyzed by metascape.Results:65 active components of Guizhi Fuling Wans were obtained by tcmsp search,mainly Quercetin,β-Sitosterol,Eburicoic acid,Polyporenic acid C,Albiflorin,etc.135 key targets of Guizhi Fuling Wan in the treatment of acute cerebral infarction were selected,including TP53,MCM2,UBC,etc.It involves 119 related signal pathways such as cell cycle,Ubiquitin mediated proteolysis and PI3K-Akt signaling pathway;Molecular docking results showed that the binding energies of several compounds with proteins were less than 8 kJ/mol.Conclusion:Guizhi Fuling Wan has the characteristics of multi-channel and multi-target in the treatment of acute cerebral infarction,which provides a scientific theoretical basis and research ideas for the future study of the mechanism of Guizhi Fuling Wan in the treatment of acute cerebral infarction.Experiment 2:Study on the mechanism of Guizhi Fuling Wan on inflammatory response and oxidative stress injury in rats with acute cerebral infarctionObjective:To observe the effect of Guizhi Fuling Wan on neurological function in ACI rats and explore its neuroprotective mechanism.Methods:The animal model of cerebral infarction was established by middle cerebral artery embolization in rats,and the therapeutic effect of Guizhi Fuling Wan was evaluated by neurological function score.HE staining and Nissl staining were performed on brain tissue to evaluate the protective effect of Guizhi Fuling Wan on neurons.The water content of brain tissue and the content of AQP4 protein were measured to evaluate the improvement effect of Guizhi Fuling Wan on brain edema.The expression of PI3K,p-Akt,Bax and Bcl-2 protein in brain tissue was determined by Western blot.The contents of TNF-α,IL-6,SOD and LDH were determined by Elisa method.The expression of TP53 and UBC was determined by q-PCR.The mechanism of action of Guizhi Fuling Wan in ACI inflammatory response,oxidative stress,apoptosis and other injuries was evaluated.Result:The results showed that the neurological function score and brain water content of the experimental group were lower than those of the model group.In the pathological section staining of the experimental group,the morphology of nerve cells was more complete than that of the model group;the expression of tp53 and UBC genes in the experimental group was lower than that in the model group.The expression of PI3K,p-Akt and Bcl2 protein in the experimental group was higher than that in the model group,and the expression of BAX protein was lower than that in the model group.The contents of TNF-α and IL-6 in the experimental group were lower than those in the model group.The activity of LDH in the experimental group was lower than that in the model group,and the activity of SOD was higher than that in the model group.Conclusion:The treatment of ACI with Guizhi Fuling Wan may be related to its ability to inhibit apoptosis,reduce inflammation and oxidative stress,and reduce brain edema.The mechanism is achieved by up-regulating the expression of PI3K/Akt pathway and Bcl2/BAX protein in brain tissue,inhibiting the expression of tp53 and UBC genes,reducing the levels of IL-6 and TNF-α,increasing the activity of SOD and reducing the content of LDH.