Withania somnifera(L.)Dunal(WS),or Ashwagandha,is used clinically against arthritis,anxiety and insomnia etc,however,its potential in aquafeeds is largely unexplored.We investigated the stress-alleviation and hepatopr...Withania somnifera(L.)Dunal(WS),or Ashwagandha,is used clinically against arthritis,anxiety and insomnia etc,however,its potential in aquafeeds is largely unexplored.We investigated the stress-alleviation and hepatoprotective mechanisms of dietary WS supplementation in adult female darkbarbel catfish.Fish were fed with WS dietary for 5 weeks,subjected to a physical stressor(saline injection),and sampled.Brain were processed for RNA-seq to identify differentially expressed genes(DEGs)linked to stress;serum were assayed for stress-and immune-related biochemical markers;livers were examined for lipid content(Oil Red O staining)and proliferation(PCNA immunos-taining).Network pharmacology were employed to construct a"compound-target"net-work:The bioactive compounds of WS were retrieved from ChEBI and PCIDB,probable targets predicted via Swiss Target Prediction,and the“WS-ingredient-target”regulatory network was constructed using Cytoscape;key ligand-protein interactions were validated by molecular docking.Findings were replicated in juvenile fish and in SG3 cells challenged with Withanolide A.Physical stress evoked 1451 DEGs in control brains,whereas WS-supplemented reduced this to 397,indicating marked mitigation of the oxid-ative-and immune-stress response.Network pharmacology and molecular docking pre-dicted that 13 bioactive compounds in WS,especially Withanolide A and Withanolide J,can interact with targets such as MAPK8,CRHR1,NR3C2,and IARS1,being associated with stress and liver function regulation respectively.Moreover,it was proved that Withanolide A could significantly modulate stress response in juveniles and enhanced pro-liferation potency of SG3 cells.Intriguingly,in theadult female darkbarbel catfish,dietary WS lowered serum cortisol,glucose,ROS,TNF-α,and IL-6(P<0.05),up-regulated hepatocyte PCNA expression,and allievated stress-induced liver injury.These findings provide novel insights and vital basis for elucidating the mechanisms whereby herbs,including WS act in stress-relief and hepatoprotection in darkbarbel catfish.And provide a framework for advancing traditional Chinese medicine(TCM)applications in aquaculture.展开更多
文摘Withania somnifera(L.)Dunal(WS),or Ashwagandha,is used clinically against arthritis,anxiety and insomnia etc,however,its potential in aquafeeds is largely unexplored.We investigated the stress-alleviation and hepatoprotective mechanisms of dietary WS supplementation in adult female darkbarbel catfish.Fish were fed with WS dietary for 5 weeks,subjected to a physical stressor(saline injection),and sampled.Brain were processed for RNA-seq to identify differentially expressed genes(DEGs)linked to stress;serum were assayed for stress-and immune-related biochemical markers;livers were examined for lipid content(Oil Red O staining)and proliferation(PCNA immunos-taining).Network pharmacology were employed to construct a"compound-target"net-work:The bioactive compounds of WS were retrieved from ChEBI and PCIDB,probable targets predicted via Swiss Target Prediction,and the“WS-ingredient-target”regulatory network was constructed using Cytoscape;key ligand-protein interactions were validated by molecular docking.Findings were replicated in juvenile fish and in SG3 cells challenged with Withanolide A.Physical stress evoked 1451 DEGs in control brains,whereas WS-supplemented reduced this to 397,indicating marked mitigation of the oxid-ative-and immune-stress response.Network pharmacology and molecular docking pre-dicted that 13 bioactive compounds in WS,especially Withanolide A and Withanolide J,can interact with targets such as MAPK8,CRHR1,NR3C2,and IARS1,being associated with stress and liver function regulation respectively.Moreover,it was proved that Withanolide A could significantly modulate stress response in juveniles and enhanced pro-liferation potency of SG3 cells.Intriguingly,in theadult female darkbarbel catfish,dietary WS lowered serum cortisol,glucose,ROS,TNF-α,and IL-6(P<0.05),up-regulated hepatocyte PCNA expression,and allievated stress-induced liver injury.These findings provide novel insights and vital basis for elucidating the mechanisms whereby herbs,including WS act in stress-relief and hepatoprotection in darkbarbel catfish.And provide a framework for advancing traditional Chinese medicine(TCM)applications in aquaculture.
