Target prediction and activity verification for the antidepressant action of Huangqin(Radix Scutellariae Baicalensis)

OBJECTIVE:To decipher the antidepressant targets and mechanisms of Huangqin(Radix Scutellariae Baicalensis)(RSB)by a novel computational system based on prediction and experimental verification.METHODS:The putative targets of RSB against depression were identified from Traditional Chinese Medicine Systems Pharmacology(TCMSP)and Drug Bank.Next,protein-protein interaction network of the anti-depression targets of RSB were identified,and differentially expressed genes(DEGs)of depression were mined from the NCBI database.Then,Kyoto Encyclopedia of Genes and Genomes and Gene Ontology were used to analysis the common targets.Finally,the selected pathways and functions were verified by experimentation.RESULTS:Thirty active compounds in RSB were predicted with high confidence by TCMSP and DrugBank,and seventy-one DEGs were identified in the GEO database.Besides,eight core target proteins were screened out by descending order of degree value,including ACHE,IL6,SLC6A4,FOS,SLC6A3,MAOB,DPP4,and JUN.These target genes were further found to be associated with pathways involved in neuronal apoptosis,such as pathways in cancer,Toll-like receptor signaling pathway,and TNF signaling.The cell proliferation assay and wound-healing assay results showed that RSB does not affect PC12 cell proliferation and chemotaxis.Unexpectedly,RSB protected PC12 cells from oxidative stress induced by H2O2 via inhibiting autophagy and apoptosis.We revealed significant changes in mice treated with 400 mg/kg RSB compared with the lipopolysaccharide mice.The possible mechanism for the antidepressive action of RSB is by reducing the expression of LC3-B in CA1 neurons.CONCLUSIONS:Our research partially expounds the mechanism of the antidepressant effect of RSB by the combination of network pharmacology prediction and experimental verification.Furthermore,it is also conducive to the application of Traditional Chinese Medicine within modern medicine.

Uncovering the mechanism of Scutellariae Radix in the treatment of breast cancer based on network pharmacology

Breast cancer(BCa)is one of the most common malignant tumors that seriously affect women’s physical and mental health and even endanger their lives.Establishing a primary prevention program for BCa remains a challenge.Traditional Chinese medicine,Scutellariae Radix(SR),has been used to treat BCa while its pharmacological mechanism is still unclear.This study aims to reveal the pharmacological mechanism of SR in the treatment of BCa.Chemical constituents of SR were obtained and the targets of the compound were identified via Traditional Chinese Medicine Systems Pharmacology Database(TCMSP).GeneCards and DrugBank databases were used to collect BCa’s related target genes.Disease-compound common target protein interaction network was established by using the STRING database.The network diagrams of the active component-action target and protein-protein interaction(PPI)networks were constructed by Cytoscape software.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were carried out to further explore the BCa mechanism and therapeutic effect of SR.As a result,121 active compounds were obtained.The Venn diagram results showed that SR and BCa had 57 intersection targets,the more frequently targeted ones are TP53,JUN,CCND1,AKT1,which could affect biological processes such as response to steroid hormone and positive regulation of cell death.KEGG analysis revealed that these targets were related to IL-17 signaling pathway,p53 signaling pathway and PI3K-Akt signaling pathway.In conclusion,this study preliminarily verified the target and mode of action of SR in the treatment of BCa,laying a foundation for further research on its mechanism.

Development of 3-mercaptopropyltrimethoxysilane(MPTS)-modified bone marrow mononuclear cell membrane chromatography for screening anti-osteoporosis components from Scutellariae Radix

Osteoporosis is a bone metabolic disease caused by the imbalance between osteoblasts and osteoclasts due to excess osteoclastogenesis,manifesting in the decrease of bone density and bone strength.Scutellariae Radix shows good anti-osteoporosis activity,but the effective component is still unclear.Cell membrane chromatography(CMC)is a biological affinity chromatography with membrane immobilized on a silica carrier as the stationary phase.It can realize a dynamical simulation of interactions between drugs and receptors on cell membrane,which is suitable for screening active compounds from complex systems.In this study,the components of Scutellariae Radix with potential anti-osteoporosis activity through inhibiting the differentiation from bone marrow mononuclear cells(BMMCs)to osteoclast were screened by a BMMC/CMC analytical system.Firstly,a new3-mercaptopropyltrimethoxysilane(MPTS)-modified BMMC/CMC stationary phase was developed to realize covalent binding with cell membrane fractions.By investigating the retention time(tR)of the positive drug,the life span of the MPTS-modified CMC columns was significantly improved from 3 to 12 days.Secondly,6 components of Scutellariae Radix were screened to show affinity to membrane receptors on BMMCs by a two-dimensional BMMC/CMC-TOFMS analytical system.Among them,tectochrysin demonstrated the best anti-osteoporosis effect in vitro,which has never been reported.We found that tectochrysin could inhibit the differentiation of BMMCs into osteoclasts induced by receptor activator of nuclear factor-κB ligand(RANKL)and macrophage colony-stimulating factor(M-CSF)in a concentration-dependent manner in vitro.In vivo,it significantly reduced the loss of bone trabeculae in ovariectomized mice,and decreased the level of C-terminal cross-linking telopeptides of type 1 collagen(CTX-1),tartrate-resistant acid phosphatase 5 b(TRAP-5 b),interleukin 6(IL-6)in serum.In conclusion,tectochrysin serves as a potential candidate in the treatment of osteoporosis.The proposed twodimensional MPTS-modified BMMC/CMC-TOFMS analytical system shows the advantages of longlife span and fast recognition ability,which is very suitable for infrequent cell lines.

Network Pharmacology Approach to Determine Active Compounds and Potential Targets Associated with the Anti-Abortion Effects of Scutellariae Radix

Background:It is widely accepted that the causes and mechanisms of abortion are very complicated.In China,Scutellariae Radix(SR)(Scutellaria baicalensis Georgi)is widely used as a traditional Chinese herbal medicine with anti-abortion effects.However,the chemical components and pharmacologic profiles of SR have not been elucidated.The network pharmacology approach can provide a system-level perspective to explore the components,targets,and mechanism of herbal medicines.Thus,this approach was employed to identify the absorbable compounds,potential targets,and signaling pathways associated with SR.Materials and Methods:In this study,we used the Lipinski rule and an oral bioavailability of>30%to identify the bioactive compounds in SR.Targets of the anti-abortion activity of SR were obtained from the PharmMapper website server database.The Search Tool for the Retrieval of Interacting Genes and DAVID databases were utilized to perform protein–protein interaction analysis and pathway enrichment analysis,respectively.Finally,Cytoscape software was used to visualize the active compound–target–Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway network of SR.Results:In total,286 chemical compounds were identified in SR;of these,27 compounds could be absorbed into the blood,and 10 compounds that had a high docking score with their corresponding targets were determined.These potentially active compounds of SR regulated 142 targets and clearly affected 29 KEGG pathways.From these targets,a total of 11 targets,which were expressed in the breast and female reproductive system,were associated with the anti-abortion effects of SR:EGFR,HRAS,HSP90 AA1,ESR1,PRKACA,SRC,GSK3 B,JAK2,IGF1 R,CDK2,and AR.In the KEGG pathway analysis,five pathways were related to the anti-abortion effect of SR,including the estrogen signaling pathway,the prolactin signaling pathway,progesterone-mediated oocyte maturation,and oocyte meiosis.Conclusions:The network pharmacology approach used in our study attempted to explain the mechanism of the anti-abortion effects of SR and has provided an alternative approach for the investigation of the effects of this complex compound.

Can Yin-Chai-Xiao-Du decoction be useful of COVID-19?the mechanism research based on network pharmacology

Background:In this study,we preliminarily investigated the mechanism of Yin-Chai-Xiao-Du decoction for the treatment of COVID-19 by the method of network pharmacology.Methods:The potential targets and pathways of Yin-Chai-Xiao-Du decoction for the treatment of COVID-19 were examined using network pharmacology;the ingredient and active targets of Yin-Chai-Xiao-Du decoction were collected from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and PharmMapper databases;the COVID-19-related targets were obtained from the online Mendelian inheritance in man,GeneCards,and GeneMANIA databases;the STRING database and Cytoscape were used to build a protein-protein interaction network,and a Network Analyzer tool was used to perform topology analysis to screen for the key ingredients and targets;the ClueGO and KOBAS 3.0 databases were for the enrichment analysis of gene function(Gene Oncology)and gene pathway(Kyoto Encyclopedia of Genes and Genomes);the herb-ingredient-target-pathway network diagram was constructed by Cytoscape.Results:The core herbs screened by the network pharmacological analysis were Jinyinhua(Lonicerae japonicae flos),Lianqiao(Forsythia suspensa),Chaihu(Bupleuri radix),Huangqin(Scutellariae radix),Yinchen(Herba Artemisiae Scopariae),Guanghuoxiang(Pogostemonis herba),Roudoukou(Semen myristicae)and Qinghao(Artemisiae annuae herba).A total of 293 active ingredients were screened by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,and the key ingredients were quercetin,kaempferol,isorhamnetin,stigmasterol,beta-sitosterol,and luteolin.Yin-Chai-Xiao-Du decoction has 138 COVID-19-related targets,and the key targets were mitogen-activated protein kinase 3,interleukin-6,tumor necrosis factor,vascular endothelial growth factor A,and CC motif ligand 2.Kyoto Encyclopedia of Genes and Genomes analysis revealed 120 enriched gene pathways,and the key pathways were signaling by interleukins,immune system,cytokine signaling in the immune system,and the signaling pathways of interleukin-17,tumor necrosis factor,and relaxin.Conclusion:The core herbs of Yin-Chai-Xiao-Du decoction are Jinyinhua(Lonicerae japonicae flos),Lianqiao(Forsythia suspensa),Chaihu(Bupleuri radix),Huangqin(Scutellariae radix),Yinchen(Herba Artemisiae Scopariae),Guanghuoxiang(Pogostemonis herba),Roudoukou(Semen myristicae)and Qinghao(Artemisiae annuae herba).The key ingredients are quercetin,kaempferol,isorhamnetin,stigmasterol,and beta-sitosterol;the critical targets are luteolin,interleukin-6,mitogen-activated protein kinase 3,tumor necrosis factor,and CC motif ligand 2;and the core signaling pathways are those mediated by interleukin-17,tumor necrosis factor,and relaxin.

Study on TLC Differentiation for Chaige Changyuan Mixture

[Objectives]This study was conducted to establish TLC methods for Scutellariae Radix,Peucedani Radix and Puerariae Radix in Chaige Changyuan Mixture.[Methods]The preparation methods of test solutions,the development conditions,the color development inspection method and other factors were investigated for thin-layer chromatography.The three main medicines of Scutellariae Radix,Peucedani Radix and Puerariae Radix were qualitatively identified.[Results]The TLC spots of Scutellariae Radix,Peucedani Radix and Puerariae Radix could be identified well by TLC,and corresponding negative control had no interference.[Conclusions]The method is simple,available and reproducible,and can be used for the quality control of Chaige Changyuan Mixture.

Comprehensive Metabolic Profiling of Modified Gegen Qinlian Decoction by Ultra-high-Performance Liquid Chromatography-Diode Array Detection-Q-Exactive-Orbitrap-Electrospray Ionization-Mass Spectrometry/Mass Spectrometry and Application of High-Performance

Objective:Gegen Qinlian decoction(GQD)is a classical traditional Chinese medicine formulation which has been used for almost 2000 years.At Guang'anmen Hospital,Beijing,a modified GQD version(m GQD)with seven instead of four herbal ingredients has been applied to treat Type 2 diabetes.Quality control is a crucial prerequisite for the therapeutic application of herbal medicines.For the identification of products derived from classical GQD,the Chinese Pharmacopeia requires the analysis of only three marker compounds.Because m GQD is a more complex mixture containing seven herbs and hundreds of constituents,the pharmacopoeia method for GQD is inadequate.Materials and Methods:A more comprehensive characterization of the formula's constituents has been developed using ultra-high-performance liquid chromatography-diode array detection(UHPLC-DAD)-Q-Exactive-mass spectrometry(MS)in electrospray ionization positive and negative mode.Moreover,a new method for the fingerprint analysis of m GQD via high-performance thin-layer chromatography(HPTLC)has been established.Results:Altogether,91 compounds have been assigned to their originating plants and 84 substances were identified either by comparison with authentic references or with data from the literature.The HPTLC method is based on the application of two different mobile phases and is able to detect both lipophilic and hydrophilic constituents of m GQD.Conclusions:The modified GQD was extensively characterized by UHPLC combined with DAD and Q-Exactive Orbitrap high-resolution MS detection,leading to the assignment and identification of compounds present in the decoction.In addition,a new method for the fingerprint analysis of the m GQD using HPTLC was established,which allows fast and simple identification of the herbal ingredients in the mixture.