Chinese traditional medicine Danshen is the radix of the perennial herbs of Salvia miltiorrhiza Bunge, which has a variety of pharmacological effects and is traditionally and extensively applied clinically to treat ca...Chinese traditional medicine Danshen is the radix of the perennial herbs of Salvia miltiorrhiza Bunge, which has a variety of pharmacological effects and is traditionally and extensively applied clinically to treat cardiovascular disorders. In this research, the genomic genes for tyrosine aminotransferase (TAT) of 38 cultivated populations of Danshen in China were cloned and bioinformatic analyses were conducted to reveal its genetic diversity and phylogeny. The full-length SmTAT was 2296 - 2444 bp including 6 exons (encoding 411 amino acids) and 5 introns. Overall, the SmTAT genes in cultivated Danshen populations are highly conserved with a relative low level of genetic diversity. The spliced exons (1236 bp) had 23 SNP variations with a rate of 1.86%, of which 22 occurred in the white flower S. miltiorrhiza Bge.f.alba population (W-SCHY-W-1) and led to 5 amino acid variations. The entire 290 SNP variations with a rate of 24% in the 5 introns occurred exclusively in W-SCHY-W-1. Phylogenetic trees based on the full-length, combined introns, the spliced exons, and the deduced amino acid sequences of SmTAT all showed a two-clade basic structure with W-SCHY-W-1 uniquely standing alone. The SmTAT gene of the white flower population (W-SCHY-W-1) is unique and especially rich in variations. The first time clarified genomic SmTAT gene structure and genetic diversity in cultivated Danshen populations laid an excellent foundation for further studies on the biosynthesis of bioactives and the molecular breeding of Danshen as well as in plant tyrosine metabolism.展开更多
Objective:To explore the effective components of Huangqi(Astragalus membranaceus(Fisch.)Bge.)and Danshen(Salvia miltiorrhiza Bge.)and predict their potential functional targets,to explore the effect and mechanism of H...Objective:To explore the effective components of Huangqi(Astragalus membranaceus(Fisch.)Bge.)and Danshen(Salvia miltiorrhiza Bge.)and predict their potential functional targets,to explore the effect and mechanism of Huangqi and Danshen ultrafine granule gel on wound vascular healing in rats by network pharmacology and experimental verification.Methods:Using the TCMSP database to screen the active components of two herbs and their potential targets,to search the target related to vascular injury through the Gene Card database,and to select the gene target with correlation coefficient greater than 10%.Then map the disease related target to the potential target of the compound,to obtain the common target,and to import the information to the String online analysis platform and Cytoscape to make the PPI map,and to carry on the topological analysis,KEGG and GO enrichment analysis through Metascape platform.The expression analysis of the main target genes of network pharmacology was verified by pathological section staining and optical density method to determine the content of vascular endothelial growth factor(VEGF)in tissues,the number of tissue new capillaries,and the wound healing rate.Results:A total of 115 active components and 297 target genes were selected from Huangqi and Danshen,and 123 target genes were intersected with vascular injury,among which IL6,AKT1,INS,VEGFA,TNF,EGF,CASP3,MAPK1,TP53 were the active genes with the highest intersection value.The pathways involved are mainly related to AGE-RAGE signaling pathway in diabetic complications,MAPK signaling pathway,Pathways in cancer,human cytomegalovirus infection,Fluid shear stress and atherosclerosis,Endocrine resistance,foxo signaling pathway etc.We experimentally validated VEGFA based on the primary target of vascular regeneration,the experimental results showed that there was different expression of VEGFA proteins in wound vascular healing process,and the quercetin,daidzein,luteolin,apigenin,tanshinone i of 115 active components were selected as the main compounds acting on the VEGFA.Conclusion:This study systematically reveals the material basis and mechanism of Huangqi and Danshen regulating wound vascular healing through multi-component,multi-target and multi-pathway and provides theoretical and scientific basis for the clinical application of Huangqi and Danshen.展开更多
文摘Chinese traditional medicine Danshen is the radix of the perennial herbs of Salvia miltiorrhiza Bunge, which has a variety of pharmacological effects and is traditionally and extensively applied clinically to treat cardiovascular disorders. In this research, the genomic genes for tyrosine aminotransferase (TAT) of 38 cultivated populations of Danshen in China were cloned and bioinformatic analyses were conducted to reveal its genetic diversity and phylogeny. The full-length SmTAT was 2296 - 2444 bp including 6 exons (encoding 411 amino acids) and 5 introns. Overall, the SmTAT genes in cultivated Danshen populations are highly conserved with a relative low level of genetic diversity. The spliced exons (1236 bp) had 23 SNP variations with a rate of 1.86%, of which 22 occurred in the white flower S. miltiorrhiza Bge.f.alba population (W-SCHY-W-1) and led to 5 amino acid variations. The entire 290 SNP variations with a rate of 24% in the 5 introns occurred exclusively in W-SCHY-W-1. Phylogenetic trees based on the full-length, combined introns, the spliced exons, and the deduced amino acid sequences of SmTAT all showed a two-clade basic structure with W-SCHY-W-1 uniquely standing alone. The SmTAT gene of the white flower population (W-SCHY-W-1) is unique and especially rich in variations. The first time clarified genomic SmTAT gene structure and genetic diversity in cultivated Danshen populations laid an excellent foundation for further studies on the biosynthesis of bioactives and the molecular breeding of Danshen as well as in plant tyrosine metabolism.
文摘Objective:To explore the effective components of Huangqi(Astragalus membranaceus(Fisch.)Bge.)and Danshen(Salvia miltiorrhiza Bge.)and predict their potential functional targets,to explore the effect and mechanism of Huangqi and Danshen ultrafine granule gel on wound vascular healing in rats by network pharmacology and experimental verification.Methods:Using the TCMSP database to screen the active components of two herbs and their potential targets,to search the target related to vascular injury through the Gene Card database,and to select the gene target with correlation coefficient greater than 10%.Then map the disease related target to the potential target of the compound,to obtain the common target,and to import the information to the String online analysis platform and Cytoscape to make the PPI map,and to carry on the topological analysis,KEGG and GO enrichment analysis through Metascape platform.The expression analysis of the main target genes of network pharmacology was verified by pathological section staining and optical density method to determine the content of vascular endothelial growth factor(VEGF)in tissues,the number of tissue new capillaries,and the wound healing rate.Results:A total of 115 active components and 297 target genes were selected from Huangqi and Danshen,and 123 target genes were intersected with vascular injury,among which IL6,AKT1,INS,VEGFA,TNF,EGF,CASP3,MAPK1,TP53 were the active genes with the highest intersection value.The pathways involved are mainly related to AGE-RAGE signaling pathway in diabetic complications,MAPK signaling pathway,Pathways in cancer,human cytomegalovirus infection,Fluid shear stress and atherosclerosis,Endocrine resistance,foxo signaling pathway etc.We experimentally validated VEGFA based on the primary target of vascular regeneration,the experimental results showed that there was different expression of VEGFA proteins in wound vascular healing process,and the quercetin,daidzein,luteolin,apigenin,tanshinone i of 115 active components were selected as the main compounds acting on the VEGFA.Conclusion:This study systematically reveals the material basis and mechanism of Huangqi and Danshen regulating wound vascular healing through multi-component,multi-target and multi-pathway and provides theoretical and scientific basis for the clinical application of Huangqi and Danshen.
