Background:Bonducellin is one of the bioactive compounds present in Caesalpinia bonduc Roxb(L).It is a homoisoflavonoid recognized for its anti-cancer,anti-androgenic,and anti-estrogenic properties and could potential...Background:Bonducellin is one of the bioactive compounds present in Caesalpinia bonduc Roxb(L).It is a homoisoflavonoid recognized for its anti-cancer,anti-androgenic,and anti-estrogenic properties and could potentially treat polycystic ovary syndrome(PCOS).However,the underlying molecular mechanism remains unexplored.This study aims to elucidate the potential molecular mechanisms of bonducellin in treating PCOS and its associated symptoms through an integrated approach combining network pharmacology,molecular docking,molecular dynamics simulation,and in vivo validation.Methods:Bonducellin-associated and PCOS-related genes were intersected using VENN analysis to determine common gene targets.KEGG pathway analysis was conducted to investigate the biological pathways involving the co-targeted genes.The protein-protein interactions of the target genes were performed to identify the key proteins interacting with bonducellin.Molecular docking and 100 ns molecular simulations were carried out to evaluate the binding affinity and conformational stability of bonducellin with the target proteins.Additionally,the acute toxicity of bonducellin was assessed on zebrafish embryos and in vivo gene expression studies were performed to examine its regulatory effect on the top co-targeted gene.Results:The intersection of bonducellin-associated and PCOS-related genes identified 76 co-targeted genes.KEGG pathway analysis revealed their involvement in 15 critical pathways,including steroid hormone biosynthesis.Protein-protein interaction and pathway enrichment analysis highlighted key targets,including MMP9,AR,KDR,PRKACA,KIT,CYP19A1,HSD11B1,ESR1,STAT3,ESR2,PRKCA,ROCK1,BRAF,HSD17B2,PIK3R1,and RAF1,all of which exhibited strong binding to bonducellin.Molecular simulations confirmed the stability of bonducellin to the top proteins,MMP9 and AR,with high binding scores.Acute toxicity studies in zebrafish embryos determined the LC50 value of bonducellin as 0.8μg/mL at 48 hpf.Gene expression analysis revealed that bonducellin differentially regulates the MMP9 gene that is involved in modulating PCOS-related pathways.Conclusion:This study suggests potential gene pathways and protein interactions through which bonducellin could exert therapeutic effects on PCOS and its associated disorders.This provides valuable insights for future research into understanding and developing bonducellin-based treatments for PCOS.展开更多
Maize(Zea mays),a major food crop worldwide,is susceptible to infection by the saprophytic fungus Aspergillus flavus that can produce the carcinogenic metabolite aflatoxin(AF)especially under climate change induced ab...Maize(Zea mays),a major food crop worldwide,is susceptible to infection by the saprophytic fungus Aspergillus flavus that can produce the carcinogenic metabolite aflatoxin(AF)especially under climate change induced abiotic stressors that favor mold growth.Several studies have used“-omics”approaches to identify genetic elements with potential roles in AF resistance,but there is a lack of research identifying the involvement of small RNAs such as microRNAs(miRNAs)in maize-A.flavus interaction.In this study,we compared the miRNA profiles of three maize lines(resistant TZAR102,moderately resistant MI82,and susceptible Va35)at 8 h,3 d,and 7 d after A.flavus infection to investigate possible regulatory antifungal role of miRNAs.A total of 316 miRNAs(275 known and 41 putative novel)belonging to 115 miRNA families were identified in response to the fungal infection across all three maize lines.Eighty-two unique miRNAs were significantly differentially expressed with 39 miRNAs exhibiting temporal differential regulation irrespective of the maize genotype,which targeted 544 genes(mRNAs)involved in diverse molecular functions.The two most notable biological processes involved in plant immunity,namely cellular responses to oxidative stress(GO:00345990)and reactive oxygen species(GO:0034614)were significantly enriched in the resistant line TZAR102.Coexpression network analysis identified 34 hubs of miRNA-mRNA pairs where nine hubs had a node in the module connected to their target gene with potentially important roles in resistance/susceptible response of maize to A.flavus.The miRNA hubs in resistance modules(TZAR102 and MI82)were mostly connected to transcription factors and protein kinases.Specifically,the module of miRNA zma-miR156b-nb-squamosa promoter binding protein(SBP),zma-miR398a-3p-SKIP5,and zma-miR394a-5p-F-box protein 6 combinations in the resistance-associated modules were considered important candidates for future functional studies.展开更多
文摘Background:Bonducellin is one of the bioactive compounds present in Caesalpinia bonduc Roxb(L).It is a homoisoflavonoid recognized for its anti-cancer,anti-androgenic,and anti-estrogenic properties and could potentially treat polycystic ovary syndrome(PCOS).However,the underlying molecular mechanism remains unexplored.This study aims to elucidate the potential molecular mechanisms of bonducellin in treating PCOS and its associated symptoms through an integrated approach combining network pharmacology,molecular docking,molecular dynamics simulation,and in vivo validation.Methods:Bonducellin-associated and PCOS-related genes were intersected using VENN analysis to determine common gene targets.KEGG pathway analysis was conducted to investigate the biological pathways involving the co-targeted genes.The protein-protein interactions of the target genes were performed to identify the key proteins interacting with bonducellin.Molecular docking and 100 ns molecular simulations were carried out to evaluate the binding affinity and conformational stability of bonducellin with the target proteins.Additionally,the acute toxicity of bonducellin was assessed on zebrafish embryos and in vivo gene expression studies were performed to examine its regulatory effect on the top co-targeted gene.Results:The intersection of bonducellin-associated and PCOS-related genes identified 76 co-targeted genes.KEGG pathway analysis revealed their involvement in 15 critical pathways,including steroid hormone biosynthesis.Protein-protein interaction and pathway enrichment analysis highlighted key targets,including MMP9,AR,KDR,PRKACA,KIT,CYP19A1,HSD11B1,ESR1,STAT3,ESR2,PRKCA,ROCK1,BRAF,HSD17B2,PIK3R1,and RAF1,all of which exhibited strong binding to bonducellin.Molecular simulations confirmed the stability of bonducellin to the top proteins,MMP9 and AR,with high binding scores.Acute toxicity studies in zebrafish embryos determined the LC50 value of bonducellin as 0.8μg/mL at 48 hpf.Gene expression analysis revealed that bonducellin differentially regulates the MMP9 gene that is involved in modulating PCOS-related pathways.Conclusion:This study suggests potential gene pathways and protein interactions through which bonducellin could exert therapeutic effects on PCOS and its associated disorders.This provides valuable insights for future research into understanding and developing bonducellin-based treatments for PCOS.
基金funded by a USDA-ARS grant in the form of a Non-Assistance Cooperative Agreement#58-6054-0-010approved for publication by the Louisiana Agricultural Experiment Station as MS#2024-306-39132.
文摘Maize(Zea mays),a major food crop worldwide,is susceptible to infection by the saprophytic fungus Aspergillus flavus that can produce the carcinogenic metabolite aflatoxin(AF)especially under climate change induced abiotic stressors that favor mold growth.Several studies have used“-omics”approaches to identify genetic elements with potential roles in AF resistance,but there is a lack of research identifying the involvement of small RNAs such as microRNAs(miRNAs)in maize-A.flavus interaction.In this study,we compared the miRNA profiles of three maize lines(resistant TZAR102,moderately resistant MI82,and susceptible Va35)at 8 h,3 d,and 7 d after A.flavus infection to investigate possible regulatory antifungal role of miRNAs.A total of 316 miRNAs(275 known and 41 putative novel)belonging to 115 miRNA families were identified in response to the fungal infection across all three maize lines.Eighty-two unique miRNAs were significantly differentially expressed with 39 miRNAs exhibiting temporal differential regulation irrespective of the maize genotype,which targeted 544 genes(mRNAs)involved in diverse molecular functions.The two most notable biological processes involved in plant immunity,namely cellular responses to oxidative stress(GO:00345990)and reactive oxygen species(GO:0034614)were significantly enriched in the resistant line TZAR102.Coexpression network analysis identified 34 hubs of miRNA-mRNA pairs where nine hubs had a node in the module connected to their target gene with potentially important roles in resistance/susceptible response of maize to A.flavus.The miRNA hubs in resistance modules(TZAR102 and MI82)were mostly connected to transcription factors and protein kinases.Specifically,the module of miRNA zma-miR156b-nb-squamosa promoter binding protein(SBP),zma-miR398a-3p-SKIP5,and zma-miR394a-5p-F-box protein 6 combinations in the resistance-associated modules were considered important candidates for future functional studies.
