Page 121 - 《水产学报》2025年第10期

李森栋，等 水产学报, 2025, 49(10): 109609

Tab. 1 Active ingredients of Withania somnifera
pubchem CID molecule name molecular formula DL OB
5280863 Kaempferol C15H10O6 0.50 0.55
5280343 Quercetin C15H10O7 0.52 0.55
193567 24-Methyldesmosterol C28H46O 0.76 0.55
173183 Campesterol C28H48O 0.59 0.55
14807783 Stigmasterone C29H46O 0.50 0.55
5281328 Fucosterol C29H48O 0.85 0.55
5280794 Stigmasterol C29H48O 0.62 0.55
222284 Beta-Sitosterol C29H50O 0.78 0.55
10494 Oleanolic acid C30H48O3 0.37 0.85
11294368 Withanolide A C28H38O6 0.78 0.55
21679022 Withanolide J C28H38O6 0.46 0.55
21679027 Withanone C28H38O6 0.45 0.55
161671 withanolide D C28H38O6 0.76 0.58

olide A exhibited strong binding affinities with the 2.5 Effects of Withanolide A on Genes’ Expres-
target, MAPK8b, CRFR1 and NR3C2, achieving sion and COR secretion in young darkbarbel
docking scores of −8.0, −10.8 and −9.8. catfish

2.4 Active Ingredients and potential Targets of To confirm the activity of Withanolide A, the
WS in fish liver in vivo model was used and young female darkbar-
bel catfish were treated by intramusclular injection
To further investigate the effects of WS on
with DMSO (as control) or Withanolide A at differ-
hepaticprotection in fish, a bioinformatics integrat-
ent dosage (1, 3, or 5 mg/kg), then mRNA levels of
ive analysis was performed using differentially
four key target genes were measured in brain and
expressed genes (DEGs) in the liver (6 864 genes)
liver by qPCR, also, the alteration of serum cortisol
and the potential target genes of WS (3 432 genes).
(COR) level were determined in fish treated by
The key regulatory factors were analyzed via the
Withanolide A. The results showed that crhr1 and
intersection of these two data sets. Likewise,
nr3c2 were prominently upregulated in all groups of
PARP1, TOP1, PRKDC, IARS1, PDK1, and MAOA
fish treated by Withanolide A, while the mapk8b
were identified as core target genes interacting with
mRNA expression was slightly inhibited, but not
the proliferating cell nuclear antigen (PCNA) (Fig.
significantly in fish treated by Withanolide A, com-
5-a,b).
pared with the fish of DMSO group (Fig. 6-a). Like-
Moreover, the molecular docking analysis was
wise, the Iars1 mRNA expression was significantly
conducted to find out the interactions between WS elevated in fish liver treated by Withanolide A at 1
and proteins. The results demonstrated that Withan- mg/kg group (Fig. 6-a). Cortisol (COR), a key
olide A, a primary bioactive compounds of WS, marker of the stress response in vertebrates, the
exhibited high binding affinities with the active site serum COR of young fish was significantly reduced
of IARS1, with binding energies of -10.8 kcal/mol by Withanolide A treatment at 1 mg/kg (P < 0.05)
(Fig. 5-c). KEGG pathway enrichment of these (Fig. 6-b). whereas the 5 mg/kg Withanolide A
intersecting targets were revealed involved in the triggered a pronounced increase of COR ( P <
Toll-like receptor signaling pathway, apoptosis, and 0.05). This biphasic effect suggests that appropriate
other stress- and proliferation-related pathways dosage of Withanolide A may suppress HPA‐axis
(Fig. 5-d). activation and mitigate stress, whereas higher

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