Based on Network Pharmacology and Experimental Verification the Mechanism of Lung Xuan Lung Turbidity Reduction Formula in Thein the Treatment of VaD was Studied
Objective The molecular mechanism of Yi Lung Xuan Lung Subduing Turbidity Formula in treating rats with vascular dementia(VaD)model was investigated by network pharmacology and animal experiments.Methods The TCMSP database platform was used to screen the active ingredients and related targets of Yi Lung,Xuan Lung and Turbidity Subduing Formula.Since Maitong could not be found in the TCMSP database,the components of Maitong were retrieved by reviewing the literature and using the BATMAN-TCM Bioinformatics Analysis Tool.The GeneCards database was used to obtain VaD-related targets and predict the potential targets of Yi Lung Xuan Lung Turbidity Reducing Formula for the treatment of VaD.The network structure of"active ingredient-target intersection"of the formula was mapped by STRING and Cytoscape 3.7.2 software,and the PPI network model was established to find the key targets.GO enrichment and KEGG pathway enrichment analyses were performed on the targets of VaD treated with Yi Lung Xuan Lung Turbidity Reducing Formula.29 rats were randomly divided into the sham-operation group,the model group,the traditional Chinese medicine group and the western medicine group.The 2-VO method was used for modelling,and the water maze was used to test the memory behaviours of the rats;HE staining was used to observe the pathological changes in the hippocampal CA1 region of the animals in each group,fluorescence detection of VEGF content in the brain of the rats in each group,and ELISA to detect the content of IL-6 and TNF-α in the brain tissues of the rats in each group;Western blot was used to detect the levels of Nrf2,HO-1,P Nrf2,HO-1,P-Akt/Akt and NF-κB.Results A total of 380 targets were obtained from Yi Lung Xuan Lung Turbidity Reducing Formula,and 183 targets were intersected with VaD;The PPI network of intersected core target genes was constructed with 92 nodes and 2610 edges.The GO results suggested that the biological processes related to VaD treatment included responses to lipopolysaccharide,oxidative stress,cell migration,etc.KEGG enrichment analysis suggested that the key pathways included NF-κB,Akt,VEGF,NOD-like receptor signalling.Compared with the model group,the number of crossing platforms in the traditional Chinese medicine group was significantly increased,the histological structure and cellular morphology of hippocampal CA1 area were intact and there were few cellular degeneration,the expression levels of Nrf2,Akt,and HO-1 were significantly increased,and the expression level of NF-κB was significantly decreased(P<0.05),and inflammatory indexes,IL-6 and TNF-α,were significantly down-regulated(P<0.01),and the immunological monoclonal fluorescence showed that the vascular endothelial cell in the traditional Chinese medicine group was significantly increased,while the activity of antioxidant factor SOD increased and the content of oxidative damage factor MDA decreased(P<0.05).Conclusion The network pharmacological analysis suggested that Yi Lung Xuan Lung and Turbidity Reducing Formula could treat VaD by regulating multiple signalling pathways and biological processes.Yi Lung Xuan Lung and Turbidity Reducing Formula increased the protein expression levels of Nrf2,P-Akt/Akt and HO-1,VEGF,and improved the degeneration of neuronal cells of hippocampal CA1 area of rats with VaD,which may be through the activation of AKT/Nrf2/HO-1,and improved the lesions of hippocampal area of VaD,inhibited oxidative damage,and decreased the content of MDA,a factor that can damage oxidative processes.lesions,inhibiting oxidative damage and down-regulating the NF-κB pathway,reducing neuroinflammation,and thus improving cognitive function in VaD model rats.
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