Integrated omics profiling of dextran sodium sulfate-induced colitic mice supplemented with Wolfberry (*Lycium barbarum*)

Inflammatory bowel disease (IBD), comprising Crohn’s disease and ulcerative colitis, is characterized by chronic gastrointestinal inflammation with symptoms such as abdominal pain, diarrhea, rectal bleeding, and weight loss. Beyond standard anti-inflammatory and immunosuppressive therapies, dietary and lifestyle interventions are commonly used. Wolfberry (Lycium barbarum) is a traditional medicinal food rich in polyphenols, carotenoids, polysaccharides, and other bioactives linked to neuroprotective, anti-aging, immune-modulating, anti-diabetic, and organ-protective effects. Despite extensive reporting on WOL constituents, few studies use whole WOL, which is how it is typically consumed. This study applies a comprehensive nutrigenomics approach (transcriptomics, proteomics, metabolomics) to elucidate how dietary supplementation with whole WOL (2%) modulates host molecular pathways in a DSS-induced colitis mouse model, examining both colon and liver to uncover mechanisms underlying potential protection against IBD.

Prior work documents multiple bioactive components of wolfberry, including polyphenols (phenylpropanoids, coumarins, lignans, flavonoids, chlorogenic acid derivatives), carotenoids (zeaxanthin, carotene), polysaccharides, betaine, cerebroside, β-sitosterol, and vitamins. Reported biological activities include ocular protection, immune enhancement, neuroprotection, anti-aging, anti-diabetic effects, protection against renal damage, and reduced immunotoxicity. Wolfberry preparations have previously reduced chemically induced colitis in mice (TNBS and DSS models). However, studies using whole fruit are limited despite its common dietary use. These findings motivate investigation of whole WOL’s effects using integrated omics to define dietary signatures and mechanisms in IBD.

Design: Male C57BL/6J mice (7 weeks old) were housed under controlled conditions and randomized into three groups (n = 6–7/group): (1) control (CON): AIN-93G diet, tap water; (2) DSS: AIN-93G diet, then 1.5% (w/v) DSS in drinking water for 9 days following 7 days on water; (3) DSSWOL: AIN-93G diet supplemented with 2% wolfberry (WOL) throughout, with 1.5% DSS administered for the final 9 days, matching DSS group timing. Pre-DSS period lasted 7 days; total DSS exposure period 9 days. WOL powder was prepared from dried wolfberry. Food, water, and DSS intakes were monitored. Outcomes: Disease activity index (DAI) calculated from daily body weight change, stool consistency, and fecal blood. At endpoint, colon length measured; mesenteric fat weight recorded; plasma collected for ELISAs of IL-6 and MMP3. Colon and liver harvested for transcriptomics, proteomics, and metabolomics. Transcriptomics: Total RNA extracted from colon and liver; RNA from each group pooled (n = 6–7 per pool). Affymetrix arrays analyzed with MAS5; genes with >1.2-fold change considered differentially expressed. RT-PCR validated selected genes (colon normalized to Rplp1; liver to Ppia). Proteomics: Total proteins from colon and liver processed for iTRAQ labeling and nanoLC-MS/MS. ProteinPilot (v4.0) with Paragon Algorithm used for ID/quantification; 95% confidence threshold; fold-change cutoffs of ≥1.2 (up) or ≤0.8 (down). Metabolomics: CE-TOFMS of polar metabolites from plasma and liver following methanol/chloroform extraction with internal standards. Peaks annotated by m/z and migration time; normalized to internal standards; concentrations estimated from standards. Data visualized via hierarchical clustering and PCA. Statistics: Data reported as mean ± SD. One-way ANOVA with Dunnett’s test; p < 0.05 considered significant versus DSS unless specified. Ethical approval obtained (University of Tokyo, Approval no. P13-739).

General/clinical: Food and water intakes were not significantly different among groups. DSS induced severe weight loss, fecal blood, diarrhea, and increased DAI. WOL supplementation reduced DAI (notably days 7–9), mitigated colon shortening, restored relative mesenteric fat weight, significantly lowered plasma MMP3, and tended to reduce plasma IL-6 (p = 0.14 vs DSS). Colon transcriptomics/RT-PCR: DSS altered 2,189 genes vs control. Versus DSS, WOL upregulated 271 and downregulated 207 genes. WOL decreased expression of proinflammatory and ECM-remodeling genes including Timp1, Ccl1, Ccl5, Il1r1, Spp1, Hp, Ptgs2, and MMPs (Mmp3, Mmp10, Mmp13), as well as Cd163 and Il6. RT-PCR confirmed DSS upregulated Mmp10, Hp, Il6, Mmp3, Timp1; WOL significantly attenuated Hp, Timp1, Mmp10 and showed trends toward lowering Il6, Cd163, Mmp3. Liver transcriptomics/RT-PCR: DSS altered 12,838 genes vs control. Relative to DSS, WOL upregulated 1,630 and downregulated 1,793 genes. WOL decreased cancer/inflammation marker genes (Saa1, Jun, S100A8) and increased fatty acid synthesis/metabolism genes (Scd1, Elovl6, Fas, Me1). RT-PCR showed DSS significantly regulated Elovl6, Fas, Jun, Mat2a, Me1, Saa1, Scd1, S100A8; WOL significantly modulated all except Jun and Saa1 (which showed suppressive trends under WOL). Proteomics (iTRAQ): Liver: 222 proteins identified; in DSSWOL vs DSS, 58 down and 75 up. WOL lowered proteins often elevated in IBD or inflammation (Calr, Trfe, Anxa5, Enob, Tthy, Sbp1, Tgm2, H4, Sodc, Cytb, Ch10, Thio, Enoa) and fibrosis-related proteins (Fibb, Fibg). Colon: 2,111 proteins detected; in DSSWOL vs DSS, 271 up and 217 down. Upregulated barrier/repair proteins included Tff3, Agr2, Cbpe, Rs3. Downregulated proteins included Shps1, Muc18, Apoe, L1cam, Ceru, Anxa3, C3, Ngal. Metabolomics (CE-TOFMS): Liver (n=131 metabolites) and plasma (n=117) profiled. PCA showed distinct clustering among CON, DSS, and DSSWOL. Ten liver and four plasma metabolites significantly altered by DSS were improved by WOL. Liver: increased glycolysis/pentose phosphate intermediates (G6P, F6P), purine metabolites (AMP, IMP, GMP), glutathione and S-lactoylglutathione, and reduced hydroxyproline (collagen degradation product). Plasma: increases observed in 2-hydroxyisobutyrate, dodecanoate, and arginine. Integrated outcome: WOL supplementation reduced colonic inflammation, preserved mucosal integrity (via reduced MMPs/TIMP1 and increased Tff3/Agr2), decreased cancer-associated markers, modulated hepatic inflammatory proteins, enhanced glycolytic recovery and fatty acid metabolism, and improved clinical indices (DAI, colon length, mesenteric fat, plasma MMP3).

The multi-omics data indicate that dietary wolfberry mitigates DSS-induced colitis by suppressing inflammatory signaling, proteases, and ECM remodeling in the colon, thereby helping maintain mucosal barrier integrity. Upregulation of Tff3 and Agr2 suggests enhanced mucus production and tight junction support, improving epithelial defense. Reduction of inflammatory and IBD-associated proteins (e.g., Muc18, Apoe, L1cam, Ceru, Shps1) and downregulation of cancer-related markers (Saa1, Jun, S100A8; Anxa3, C3) may reduce risk factors for colitis-associated carcinogenesis. In the liver, WOL reversed inflammatory protein signatures and restored metabolic pathways, with increased glycolytic intermediates (G6P, F6P) and fatty acid metabolism gene expression (Fas, Me1, Scd1, Elovl6), potentially supporting energy demands during inflammation and contributing to improved body weight. The intervention also appears to counteract early events preceding overt IBD pathology, such as DSS-associated suppression of hepatic Scd1 potentially driven by colonic cytokines (e.g., IL-6). Collectively, these findings support whole WOL as a dietary strategy to ameliorate inflammation, maintain gut barrier function, and potentially lower progression risks in IBD.

A 2% whole wolfberry-supplemented diet ameliorated DSS-induced colitis in mice. Integrated colon transcriptome/proteome data showed reduced inflammation and preserved mucosal wall integrity with increased mucus/epithelial defense proteins. Hepatic transcriptome/proteome/metabolome data indicated reduced inflammatory markers and recovery of glycolysis and fatty acid metabolism, aligning with improved clinical indices. These results suggest daily inclusion of wolfberry could help maintain gut health and reduce disease severity in mucosal damage-associated conditions. Future work should further evaluate long-term efficacy, dose-response, and translation to human IBD.

The DSS administration period was relatively short. RNA for microarray was pooled within groups (n = 6–7), which limits assessment of inter-individual variability. The study used only male C57BL/6J mice in a chemical colitis model, which may constrain generalizability.