Exploring the mechanism of Huangjing pill in the treatment of Alzheimer’s disease based on network pharmacology and molecular docking

Objective To reveal the mechanism of Huangjing pill in treating Alzheimer’s disease(AD)based on network pharmacology and molecular docking technology.Methods We obtained the active ingredients and targets of Huangjing pill through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,and supplemented the effective components by consulting literature and predicted targets through the PharmMapper database.We used DrugBank,the GeneCards,the TTD,and the OMIM database to collect targets of AD.The Venn diagram was drawn and the key targets of Huangjing pill in the treatment of AD were obtained by Venny 2.1 platform.The Cytoscape 3.8.1 software was used to construct a network diagram of“drugs-active ingredients-key targets-disease”.The protein-protein interaction(PPI)network diagram was constructed through the STRING 11.5 database.DAVID database was used for Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis.AutoDock Vina1.1.2 software was used for molecular docking of the active components and core targets,and PyMOL 1.7.2.1 software was used for visual processing.Results After screening,we obtained 13 active ingredients and 116 targets of Huangjing Pill,1438 related targets for AD,and 75 common targets.566 items by GO enrichment analysis and 149 items related to KEGG pathway enrichment were obtained.Molecular docking results showed that there is a strong affinity between the key active ingredients and the core targets.Conclusion This study revealed that Huangjing pill could treat AD through multiple components,multiple targets and multiple pathways.

Differential distribution of authenticity results of different varieties of refined honey based on stable isotope ratio method

Objective Using the stable isotope ratio method for the authenticity identification and variety identification of refined honey.Methods In this paper,a total of 17 samples of different varieties of refined honey were used to obtain refined honey proteins by precipitation with sodium tungstate solution and sulphuric acid solution.The isotope mass spectrometer was used to simultaneously detect theδ^(13)C values of refined honey proteins and refined honey as well as theδ^(18)O andδ^(2)H values of refined honey,processed of the results obtained,analysed the authenticity of the samples and conduct a variety identification study.Results Tested of the resulting honey samples,the results showed that four batches of refined honey did not up to standard,two batches of C-4 vegetable syrup were detected as adulterated,and two batches of protein were not detected.Theδ^(18)O andδ^(2)H values of refined honey were also found to be effective in distinguishing the varietal origin of refined honey to a certain extent.Conclusions The stable isotope ratio method is useful in the authenticity identification of refined honey,and provides new ideas to further promote the authenticity of refined honey and variety identification research.

Effect of Toosendan Fructus before and after stir-frying process extract on CYP450 enzyme activities in rats in vivo and vitro by cocktail probe drug method

Background:The traditional Chinese medicine Toosendan Fructus has certain hepatotoxicity,which is used after being processed by stir-frying to attenuate toxicity.However,there are few studies on its attenuating toxicity mechanism.The effects of Toosendan Fructus on the activities of CYP450P1A2,CYP2E1 and CYP3A4 were studied in vivo and in vitro and the dose-toxicity mechanism of hepatotoxicity before and after stir-frying was explored to provide the basis for safe,rational use of Toosendan Fructus.Methods:The rat liver microsomes in vitro incubation method and in vivo pharmacokinetics were used to detect the concentrations of phenacetin,chlorzoxazone and dapsone in the liver microsomes in vitro incubation system and the rat plasma to study the effect of stir-frying of Toosendan Fructus on the activity of CYP450P1A2,CYP2E1,CYP3A4.Results:The results of pharmacokinetics in vivo showed that the AUC of phenacetin and dapsone in different groups was lower,and CL value was higher than those of the normal group.At the same dose,the AUC of stir-fried Toosendan Fructus was higher than that of the raw,while CL value was lower.For the same processed product,AUC value was high-dose>low-dose>middle-dose group,CL value was high-dose<low-dose<middle-dose.AUC and CL values of chlorzoxazone showed no difference from those of the normal group.The results of pharmacokinetics in vivo showed that Toosendan Fructus can induce the activity of CYP3A4 in a dose-dependent manner and the induction effect will decrease after stir-frying in vitro.Conclusion:The toxicity attenuation of Toosendan Fructus may be related to the decrease of induction effect after stir-frying.These results would provide the basis for safe,rational use of Toosendan Fructus.

Effects of Euphorbiasteroid on gene expression in lung cancer cells based on gene chip

Objective Using gene chip technology to explore the molecular mechanism of euphorbiasteroid in the treatment of non-small cell lung cancer(NSCLC).Methods A549 cells were used as the in vitro model,and they were randomly divided into control group and different concentrations of euphorbiasteroid administration groups.Each group had 3 duplicate wells,after the cells were cultured in vitro,the cell viability was evaluated by CCK-8 method.Gene chip technology was used to screen the differentially expressed genes(DEGs)between the control group and the euphorbiasteroid administration group.The differential genes were further analyzed for Gene Ontology(GO)function enrichment and Kyoto Encyclopedia of Genes and Genome(KEGG)pathway enrichment analysis.The STRING online analysis platform combined with Cytoscape software to construct a target protein interaction(PPI)network and perform topological analysis to screen key targets,and use Real-time PCR(RT-PCR)and molecular docking technology to verify key targets.Results According to the analysis of gene chip data,276 differentially expressed genes were screened,including 117 up-regulated genes and 159 down-regulated genes.GO analysis showed that differentially expressed genes were mainly involved in cell division,cell proliferation,cell cycle and other processes,involving protein binding,protein kinase binding and other functions,and were mainly distributed in nucleoplasm,chromosomes and other parts.KEGG signaling pathway analysis showed that differentially expressed genes were involved in cell cycle,pyrimidine metabolism,p53 signaling pathway and other pathways.PPI network analysis showed that CCNA2,TOP2A,CCNB1,CDC20,and RRM2 may be the key targets of euphorbiasteroid in the treatment of NSCLC.RT-PCR results showed that the expressions of CCNA2,TOP2A,CCNB1,CDC20,and RRM2 were significantly down-regulated in the euphorbiasteroid administered group,which was consistent with the gene chip results.Molecular docking results showed that euphorbiasteroid had good affinity with key targets and could bind spontaneously and stably.Conclusion The combination of gene chip,RT-PCR technology and molecular docking technology can find out the differential genes after the intervention of euphorbiasteroid,which can be used to explore the mechanism of euphorbiasteroid in the treatment of NSCLC.

Key genes of deficiency of liver and kidney type chronic pain through bioinformatics analyses

Objective To screen the key genes of chronic pain and provide a reference for the treatment of chronic pain.Methods We performed comprehensive bioinformatics analysis by screening chronic primary pain-related datasets to obtain differentially expressed genes(DEGs)and then imported DEGs into the DAVID database for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Gene Set Enrichment Analysis(GESA)analysis was done by GSEA_4.1.0 software.At the same time,we imported the intersecting genes into the STRING database and processed them by Cytoscape_3.8.1 software to obtain the protein-protein interaction(PPI)network and the central gene.Results As a result,a total of 54 DEGs were screened,including 47 up-regulated genes,1 down-regulated gene,and 6 genes that were expressed differently in different datasets.23 GO terms and 8 KEGG pathways were enriched by DAVID.PPI network analysis found that SPI1,STAT3,TNFRSF1B,PTGS2,and CXCL1 genes interacted more strongly with other genes,and were predicted to be key genes in chronic primary pain.Conclusion Our results suggested that 5 DEGs,STAT3,SPI1,TNFRSF1B,PTGS2,and CXCL1,have the potential to be used as prognostic and predictive markers for the clinical management of patients with this disease.

Effects of different processing methods on the quality of Polygonatum cyrtonema Hua

Background:In this study,we explored the effects of different processing methods on the quality of Polygonatum cyrtonema Hua(PC),and the role of Huangjiu in the processing procedure.Methods:The sensory characteristics of the crude product,steamed product,and wine-processed product of PC were described.The colorimeter was used to analyze the chromatic values of three different processed products on PC.At the same time,the contents of the water extract and alcohol extract were measured separately.The content of three different processing Polygonatum Polysaccharide(PCP)was determined using 0.2%anthrone-sulfuric acid.The correlation difference between the chromatic values and chemical composition of different PC products was analyzed using various analytical methods.Results:The surface colors gradually deepened,the sweetness increased,the viscosity strengthened,and the tongue-numbing sensation disappeared after PC processing.The contents of extract and L*gradually decreased from the crude to the steamed to the wine-processed product,consistent with the pattern of surface color alteration.While,E*ab gradually increased.The content of PCP was crude product>wine-processed product>steamed product.The results of multivariate statistical analysis showed that the samples processed for crude,steamed,and wine-processed product were clustered into three classes.The correlation analysis showed that L*and E*ab were highly significant positively correlated with the content of PCP,and a*was significantly negatively correlated with the content of PCP.Conclusion:The results showed that the wine-processed product had the best quality.The internal quality of the PC was correlated with its characteristics and chromatic value.In this study,we investigated the internal and external quality of three different products of PC in order to provide a reference for further research on the impact of different processing methods on PC quality,the standardization of PC processing,and the role of Huangjiu in the processing of PC.

Study on the regulatory effect of liver X receptor in HEK293 cells by six main diterpene esters in Semen Euphorbiae

Background:To study the effects of the main diterpene esters in Euphorbia factor L_(1),L_(2),L_(3),L_(7a),L_(7b)and L_(8)on the transcriptional activity and protein expression of liver X receptor(LXR).Methods:The effect of the main diterpene ester components in Semen Euphorbiae on the viability of HEK293 cells were studied by MTT assay.The LXR-Luc plasmid vector was transfected into HEK293 cells and treated with Euphorbia factor L_(1),L_(2),L_(3),L_(7a),L_(7b)and L_(8)for 24 h.The effect of the main diterpene ester components of Semen Euphorbiae on LXR-Luc luciferase activity was investigated by dual luciferase reporter gene system,and the expression of LXRαprotein was detected by Western Blot.Results:Euphorbia factor L_(1),L_(2),L_(3),L_(7a),L_(7b)and L_(8)could significantly reduce the relative luciferase activity(RLU)of LXRα,and the expression level of LXRαprotein was significantly down-regulated.Conclusion:Euphorbia factor L_(1),L_(2),L_(3),L_(7a),L_(7b)and L_(8)can inhibit the expression of LXR protein level,which may be achieved by inhibiting the transcriptional activity of LXR.

Scorpion Venom Heat-Resistant Peptide is Neuroprotective against Cerebral Ischemia-Reperfusion Injury in Association with the NMDA-MAPK Pathway

Scorpion venom heat-resistant peptide(SVHRP)is a component purified from Buthus martensii Karsch scorpion venom. Our previous studies have shown that SVHRP is neuroprotective in models of Alzheimer’s disease and Parkinson’s disease. The present study aimed to explore the potential neuroprotective effects of SVHRP on cerebral ischemia/reperfusion(I/R) injury, using a mouse model of middle cerebral artery occlusion/reperfusion(MCAO/R) and a cellular model of oxygen-glucose deprivation/reoxygenation(OGD/R). Our results showed that SVHRP treatment decreased the neurological deficit scores, edema formation, infarct volume and neuronal loss in the MCAO/R mice, and protected primary neurons against OGD/R insult. SVHRP pretreatment suppressed the alterations in protein levels of N-methyl-D-aspartate receptors(NMDARs) and phosphorylated p38 MAPK as well as some proinflammatory factors in both the animal and cellular models. These results suggest that SVHRP has neuroprotective effects against cerebral I/R injury, which might be associated with inhibition of the NMDA-MAPKmediated excitotoxicity.