Study On The Effect And Mechanism Of Qi Fu Yin In Treating Vascular Dementia Based On Network Pharmacology

Objective: Based on efficacy network of Qi Fu Yin in treating vascular dementia,we studied the protective effect and potential mechanism of Qi Fu Yin on VD rats and SAM mice.Methods: 1.We comprehensively reviewed articles from various databases,including CNKI,Wanfang,VIP,CBM,Pub Med and Web of Science published before June 2020,for all randomized controlled trials on dementia treatment with QFY.Then,we selected eligible literatures,extracted related data,and assessed risk of bias.Forest plots of total clinical effective rate,MMSE score,HDS score and ADL score illustrated the difference between the experimental group and the control group.Finally,publication bias was further analyzed using funnel plot,sensitivity analysis,begg and Egger's test.2.Using the method of network pharmacology,multiple databases were combined used to analyze the active ingredients,targets of Qi Fu Yin and vascular dementia.A network diagram of “QFY active ingredients—action targets of VD” was constructed.We further carried out the network topology analysis to screen the core action targets,which then analyzed by Cluo Go and Pathway analysis to predict the possible mechanism of action.3.We determined optimal chromatographic conditions to proceed specificity,linear relationship and methodological study.The HPLC fingerprint analysis method of Qi Fu Yin was established.Moreover,the quality control of Qi Fu Yin extracted from ten batches were performed to ensure the consistency of subsequent pharmacodynamics experiments.4.Vascular dementia model was established by ligating bilateral common carotid arteries of SD rats.After 6 weeks of QFY administration,the effects of QFY on the learning and memory of VD rats were observed through Morris water maze,new object recognition and the passive avoidance response.The hippocampus,striatum,and corpus callosum were used as observation objects to explore the effects of QFY on the protein expression of MMP2,MMP9,ZO-1,Occludin,PSD95,CX47,and MBP in each group of rats.The effects of QFY on brain tissue pathology were observed by Nissl staining and LFB myelin staining.5.The effect of QFY on the learning and memory of SAMP8 mice were observed through Y maze,new object recognition and nesting experiments after 30 days of QFY administration.Taking hippocampus and cortex as observation objects,we explored the effects of QFY on MMPs-related regulatory proteins(MMP2,MMP9,ZO-1,Occludin,PSD95),oxidative stress markers(SOD,MDA),inflammation(NF-?B,IL-1?,IL-6)and apoptosis related regulatory protein expression(Bcl-2,Bax,Cyt C).In addition,the effect of QFY on the pathology of SAMP8 mice brain tissue was observed by HE staining and TUNEL immunofluorescence.Results:1.Finally,9 RCTs,involving 697 patients,were included in this study.The results of Meta-analysis confirmed the efficacy and safety of QFY in the clinical treatment of dementia.2.We confirmed several active ingredients such as quercetin,isoglycyrol,glabridin,quercetin,Phaseolinisoflavan and ?-sitosterol in Qi Fu Yin,who may act on targets such as MMP2,MMP3 and MMP9 etc and play anti-vascular dementia effect through possible signal pathways such as extra-nuclear estrogen signaling,collagen degradation,and activation of matrix metalloproteinase.3.We established the HPLC QFY fingerprint using acetonitrile—0.1% phosphoric acid solution as mobile phase for the gradient elution.The results of specificity,linear relationship and methodological study all met fingerprint requirements.Eighteen common peaks were identified.Moreover,ferulic acid,ginsenoside Rb1 and glycyrrhizic acid were classified as three index components.Ten batches of QFY fingerprint revealed the similarity ranged from0.953 to 1.000 and the contents of ferulic acid ranged from 298.59 to 311.29 mg/kg,ginsenoside Rb1 ranged from 511.04 to 629.23 mg/kg,and glycyrrhizic acid ranged from701.05 to 890.62 mg/kg.The results indicated that QFY had a good stability.4.Compared with sham group,We found that the water maze escape latency increased,the number of platform crossings and platform stay time in the target quadrant of VD rats decreased;the new object recognition index and residence time of model rats decreased;the model rats entered the dark box more times and stay for a long time.QFY administration ameliorated all above behavioral changes.QFY can down-regulate the expression of MMP2 protein,up-regulate the expression levels of ZO-1,Occludin,PSD95,CX47,MBP protein,and improve the pathological changes of the hippocampus and white matter of VD rats.5.Compared with SAMR1 mice,spontaneous alternate response rate,new object recognition index and residence time,nesting score of SAMP8 mice were significantly reduced.After QFY intervention,the above behavioral indicators can be improved.QFY can down-regulate the expression of MMP2 and MMP9 proteins in the cortex;increase the activity of SOD and reduce the content of MDA;down-regulate the expression of NF-?B,IL-1?,IL-6,Bax,and Cyt C proteins;and improve the pathological changes in the brain tissue of SAMP8 mice.Conclusion:1.The VD model was established through permanent ligation of bilateral common carotid arteries,characterized by cerebral ischemia and deficiency of Qi and blood.In traditional Chinese medicine,the pathogenesis of VD was consistent with insufficiency of Qi and blood,poor blood flow,and impaired brain obstruction.QFY can improve the learning and memory of VD rats by invigorating qi and strengthening spleen,invigorating the kidney and cultivating blood,promoting blood circulation and resuscitation.2.Starting with the activation of matrix metalloproteinases in the efficacy network study on QFY in treating VD,We verified that the mechanism of QFY on VD rats is related to the regulation of MMPs and improvement of blood-brain barrier.3.SAMP8 mice,characterized by deficiency and aging of internal viscera,which accord with deficiency of Qi and blood,brain marrow deprived of nourishment and loss of function of mind in the pathogenesis and etiology of VD.QFY can improve the learning and memory of SAMP8 mice.4.The mechanism of QFY on SAMP8 mice may be related to the regulation of MMPs,improvement of oxidative stress,inflammation,apoptosis and so on.