Page 118 - 《水产学报》2025年第10期

李森栋，等 水产学报, 2025, 49(10): 109609

(ImageJ V1.8.0.112, USA). Since PCNA protein fied in the CW group, including 752 upregulated
is usually considered as a cell proliferation and 699 downregulated genes (Fig.2-a). In contrast,
marker, the proliferation potency of cells was the NC group showed only 397 DEGs (170 upregu-
always assessed as following: the proliferation lated, 227 downregulated), indicating less pro-
rate = the number of cells with PCNA positive nounced transcriptomic disturbance under WS sup-
signals / the total number of cells •100%. plementation (Fig.2-b). GO enrichment analysis

showed that DEGs in CW vs. CC were significantly
1.12 Statistical analysis
enriched in biological processes such as protein
Statistical analyses were conducted using folding, response to stimulus, signal transduction,
GraphPad Prism version 10 (GraphPad Software, and immune system process. KEGG pathway ana-
San Diego, CA, USA). Data are presented as mean ± lysis revealed that the DEGs were significantly
standard error of the mean (SEM). One-way ana- enriched in MAPK signaling pathway, HIF-1 sig-
lysis of variance (ANOVA) was employed for mul- naling pathway, NOD-like receptor signaling path-
tiple group comparisons, followed by Tukey’s post- way, oxidative phosphorylation, and neuroactive
hoc test for pairwise comparisons. For comparisons ligand–receptor interaction.
between two groups, an unpaired t-test was applied. Notably, several classical stress-related genes
A P-value of less than 0.05 was considered statistic- were markedly upregulated in the CW group. These
ally significant. included hsp70 (log FC = 2.21), hsp90aa1.1
2
(log FC = 1.82), sod1 (log FC = 1.63), and il1b
2
2
2 Results
(log FC = 2.04), indicating activation of the heat
2

shock response and inflammatory signaling. Mean-
2.1 Differential Expressed Genes induced by
while, pro-apoptotic genes such as bax (log FC =
2
stress stimuli in Brain Tissue
1.34) and casp9 (log FC = 1.21) were also signific-
2
To determine the molecular alterations in antly upregulated (Fig.2-c, d). In contrast, in the NC
brain tissues during responding to acute stress and group, these genes exhibited much lower fold
effects of dietary WS on stress response in darkbar- changes or remained unchanged, suggesting a sup-
bel catfish, RNA-seq analysis was performed on pressive effect of WS on stress-induced gene activa-
brain tissues from control (CC), acute stress (CW), tion.

and WS-supplemented (NC) groups. A total of
2.2 Gene Ontology and Pathway Enrichment
57.55 million reads were generated prior to filter-
Analysis
ing. The data exhibited high quality, with a Q20
The distinct gene expression patterns across
base percentage of 97.99% and a Q30 base percent-
the four experimental groups (CC, CP, NC, and NP)
age of 94.07%. The GC content was approximately
43.72%. After filtering, the average read length for were illustrated with a heatmap (Fig. 3-a). Through
both ends was 149 bp, and the duplication rate was the GO enrichment analysis of the DEGs from con-
calculated at 3.72%. The peak insert size was 259 trol groups without WS dietary (CP vs. CC) (Fig. 3-b),
bp. Overall, the sequencing data demonstrated high the Go terms were significantly enriched in categor-
quality, with substantial reductions in sequencing ies related to synaptic and trans-synaptic signaling.
artifacts after filtering, ensuring the reliability of Additionally, the DEGs were also mainly involved
subsequent analyses. To validate the RNA-seq data, in the biological process associated with blood
qPCR was performed for selected genes with the coagulation and the response to wounding. The
gene specific primers (Table S2). KEGG pathway analysis showed of the DEGs from
Compared with the CC group, a total of 1 451 control groups without WS dietary (CP vs. CC),
differentially expressed genes (DEGs) were identi- were mainly involved in the calcium signaling path-

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