Saline-alkali stress is a significant abiotic stressor that affects the growth of tomato(Solanum lycopersicum L.).Our previous studies reported that the homologous genes SIWRKY80 and SIWRKY81 can both resist saline-al...Saline-alkali stress is a significant abiotic stressor that affects the growth of tomato(Solanum lycopersicum L.).Our previous studies reported that the homologous genes SIWRKY80 and SIWRKY81 can both resist saline-alkali stress by mediating the JA pathway.In this study,the cooverexpressing(SOE)and co-silencing(SQ)lines of SlWRKY80 and SIWRKY81 to saline-alkali treatment,and the results showed that SOE line had significantly higher morphological indices,physiological indices,and endogenous JA and MeJA contents than the WT,while SQ line was opposite.In addition,the SOE line showed a significant increase in autophagy against saline-alkali stress,and both SlWRKY80 and SlWRKY81 were positively regulating the transcription of the autophagy gene SIATG13b as screened and verified by transcriptome screening,qRT-PCR,WB(Western Blot),Y1H(Yeast One-Hybrid),ChIP-qPCR,EMSA(Electrophoretic Mobility Shift Assay),and D-LUC(D-Luciferin Experiment),and also had a synergistic effect in activating SIATG13b.The growth and physiological indexes of VIGS-SIATG13b line were significantly lower than those of other groups,and pro-SlATG13b is significantly activated by MeJA and saline-alkali signals,so the SIATG13b was regulating the resistance of tomato to saline-alkali stress.In summary,the increase of endogenous JA and MeJA content in tomato after saline-alkali stress inhibited the expression and synthesis of SIJAZ1,activated the transcription and protein synthesis of SlWRKY80 and SlWRKY81,and SlWRKY80 and SlWRKY81 coordinately regulated the transcription of SlATG13b,which resulted in the production of more autophagosomes and enhanced the resistance of tomato to saline-alkali stress.This study further improved the molecular mechanism of SlWRKY80 and SIWRKY81 mediating the JA pathway against saline-alkali stress,and laid a solid theoretical foundation for tomato saline-alkali resistance breeding.展开更多
Objective:To explore the molecular mechanism and action pathways of Wumei Pill in the treatment of autoimmune hepatitis(AIH)using network pharmacology and molecular docking methods.Methods:The active components and ta...Objective:To explore the molecular mechanism and action pathways of Wumei Pill in the treatment of autoimmune hepatitis(AIH)using network pharmacology and molecular docking methods.Methods:The active components and targets of Wumei Pill,as well as AIH-related disease targets,were screened through the TCMSP,GeneCards,OMIM,and Disgenet databases.Cytoscape 3.9.1 was used to construct a series of topological networks,followed by Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis and Gene Ontology(GO)enrichment analysis.Molecular docking and visualization were performed using AutoDockTools 1.5.7 and PyMOL 2.4.0.Results:A total of 124 active components of Wumei Pill,877 drug targets,1130 disease targets,and 64 overlapping targets were obtained.GO enrichment analysis yielded 82 biological processes,4 cellular components,and 19 molecular functions.KEGG pathway enrichment analysis identified 21 signaling pathways.Conclusion:Wumei Pill can act on targets such as Tumor Necrosis Factor(TNF),Caspase 3(CASP3),C-X-C Motif Chemokine Ligand 8(CXCL8),Nuclear Factor Kappa B Subunit 1(NFKB1),and Transforming Growth Factor Beta 1(TGFB1)through active components including girinimbine and(R)-tetrahydroberberine.It further regulates inflammation and apoptosis-related pathways such as tumorrelated signaling pathways and Th17 cell differentiation pathway to treat AIH.This study provides a theoretical basis for in-depth research on the mechanism of Wumei Pill in the treatment of AIH and the development of therapeutic drugs.展开更多
基金supported by the 2024 Chongqing Postdoctoral Innovative Talent Support Program Project(Grant No.CQBX202420)China Agriculture Research System(CARS-23-B08)+2 种基金Fundamental Research Funds for the Cornell University(Grant No.SWU-KF25027)Key Research and Development Projects of Shaanxi Province(Grant No.2024NC-ZDCYL-05-08)Northwest A&F University National College Students Innovation and Entrepreneurship Training Program(Grant No.202410712015)。
摘要Saline-alkali stress is a significant abiotic stressor that affects the growth of tomato(Solanum lycopersicum L.).Our previous studies reported that the homologous genes SIWRKY80 and SIWRKY81 can both resist saline-alkali stress by mediating the JA pathway.In this study,the cooverexpressing(SOE)and co-silencing(SQ)lines of SlWRKY80 and SIWRKY81 to saline-alkali treatment,and the results showed that SOE line had significantly higher morphological indices,physiological indices,and endogenous JA and MeJA contents than the WT,while SQ line was opposite.In addition,the SOE line showed a significant increase in autophagy against saline-alkali stress,and both SlWRKY80 and SlWRKY81 were positively regulating the transcription of the autophagy gene SIATG13b as screened and verified by transcriptome screening,qRT-PCR,WB(Western Blot),Y1H(Yeast One-Hybrid),ChIP-qPCR,EMSA(Electrophoretic Mobility Shift Assay),and D-LUC(D-Luciferin Experiment),and also had a synergistic effect in activating SIATG13b.The growth and physiological indexes of VIGS-SIATG13b line were significantly lower than those of other groups,and pro-SlATG13b is significantly activated by MeJA and saline-alkali signals,so the SIATG13b was regulating the resistance of tomato to saline-alkali stress.In summary,the increase of endogenous JA and MeJA content in tomato after saline-alkali stress inhibited the expression and synthesis of SIJAZ1,activated the transcription and protein synthesis of SlWRKY80 and SlWRKY81,and SlWRKY80 and SlWRKY81 coordinately regulated the transcription of SlATG13b,which resulted in the production of more autophagosomes and enhanced the resistance of tomato to saline-alkali stress.This study further improved the molecular mechanism of SlWRKY80 and SIWRKY81 mediating the JA pathway against saline-alkali stress,and laid a solid theoretical foundation for tomato saline-alkali resistance breeding.
基金University-Level College Student Innovation and Entrepreneurship Training Program of Qiqihar Medical UniversityExploring the Mechanism of Ganjiang Huangqin Huanglian Renshen Decoction on AIH Mice Based on Network Pharmacology and In Vivo Experiments(Project No.:X202311230039)。
摘要Objective:To explore the molecular mechanism and action pathways of Wumei Pill in the treatment of autoimmune hepatitis(AIH)using network pharmacology and molecular docking methods.Methods:The active components and targets of Wumei Pill,as well as AIH-related disease targets,were screened through the TCMSP,GeneCards,OMIM,and Disgenet databases.Cytoscape 3.9.1 was used to construct a series of topological networks,followed by Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis and Gene Ontology(GO)enrichment analysis.Molecular docking and visualization were performed using AutoDockTools 1.5.7 and PyMOL 2.4.0.Results:A total of 124 active components of Wumei Pill,877 drug targets,1130 disease targets,and 64 overlapping targets were obtained.GO enrichment analysis yielded 82 biological processes,4 cellular components,and 19 molecular functions.KEGG pathway enrichment analysis identified 21 signaling pathways.Conclusion:Wumei Pill can act on targets such as Tumor Necrosis Factor(TNF),Caspase 3(CASP3),C-X-C Motif Chemokine Ligand 8(CXCL8),Nuclear Factor Kappa B Subunit 1(NFKB1),and Transforming Growth Factor Beta 1(TGFB1)through active components including girinimbine and(R)-tetrahydroberberine.It further regulates inflammation and apoptosis-related pathways such as tumorrelated signaling pathways and Th17 cell differentiation pathway to treat AIH.This study provides a theoretical basis for in-depth research on the mechanism of Wumei Pill in the treatment of AIH and the development of therapeutic drugs.