Journal: Frontiers in Microbiology

Article Title: Lily polysaccharides alleviate colitis through the microbiota–N8-acetylspermidine–cGAS–STING signaling axis

doi: 10.3389/fmicb.2025.1686902

Figure Lengend Snippet: The therapeutic effect of lily polysaccharide on colitis. (a) Changes of mice weight. (Day x/Day 0 *100%, n = 5). (b) Disease activity index. ( n = 4). (c,d) Quantified lengths of colonic tissues isolated from mice after treatment ( n = 5). (e) H&E-stained histological sections of colons. Representative digital photos. (f) The pathology score of colons from mice in the different treatment group ( n = 5). (g–i) The levels of pro-inflammatory cytokines IL-1 β , IL-6 and TNF- α in colon of mice were detected by ELISA ( n = 5). (j) The expression levels of MUC2 were determined by immunofluorescence staining in the mouse colon. Representative digital photos. Data are presented as the mean ± SEM. Statistically significant differences are indicated; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the samples were washed again with PBST and incubated overnight at 4 °C with a primary antibody against MUC2 (Rabbit polyclonal IgG, Proteintech, Catalog No. 27675-1-AP, unconjugated; 1:200 dilution).

Techniques: Activity Assay, Isolation, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence

Journal: Frontiers in Microbiology

Article Title: Lily polysaccharides alleviate colitis through the microbiota–N8-acetylspermidine–cGAS–STING signaling axis

doi: 10.3389/fmicb.2025.1686902

Figure Lengend Snippet: Fecal bacteria transplantation experiments showed that LP could positively regulate the gut microbiota. (a) Changes of mice weight. (Day x/Day 0 *100%, n = 5). (b) Disease activity index ( n = 5). (c,d) Quantified lengths of colonic tissues isolated from mice after treatment ( n = 5). (e) H&E-stained histological sections of colons. Representative digital photos. (f) The pathology score of colons from mice in the different treatment group ( n = 5). (g–i) The levels of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in colon of mice were detected by ELISA ( n = 5). (j) The expression levels of MUC2 were determined by immunofluorescence staining in the mouse colon. Representative digital photos. Data are presented as the mean ± SEM. Statistically significant differences are indicated; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the samples were washed again with PBST and incubated overnight at 4 °C with a primary antibody against MUC2 (Rabbit polyclonal IgG, Proteintech, Catalog No. 27675-1-AP, unconjugated; 1:200 dilution).

Techniques: Bacteria, Transplantation Assay, Activity Assay, Isolation, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence

Journal: Frontiers in Microbiology

Article Title: Lily polysaccharides alleviate colitis through the microbiota–N8-acetylspermidine–cGAS–STING signaling axis

doi: 10.3389/fmicb.2025.1686902

Figure Lengend Snippet: Replenishing N8AS can effectively alleviate enteritis. (a) Changes of mice weight.(Day x/Day 0 *100%, n = 5). (b) Disease activity index. ( n = 5). (c,d) Quantified lengths of colonic tissues isolated from mice after treatment ( n = 5). (e) H&E-stained histological sections of colons. Representative digital photos. (f) The pathology score of colons from mice in the different treatment group ( n = 5). (g–i) The levels of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α in colon of mice were detected by ELISA ( n = 5). (j) The expression levels of MUC2 were determined by immunofluorescence staining in the mouse colon. Representative digital photos. (k–o) The protein expression levels of cGAS, STING, p-TBK, p-IRF3 were assessed by western blot analysis. Data are presented as the mean ± SEM. Statistically significant differences are indicated; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the samples were washed again with PBST and incubated overnight at 4 °C with a primary antibody against MUC2 (Rabbit polyclonal IgG, Proteintech, Catalog No. 27675-1-AP, unconjugated; 1:200 dilution).

Techniques: Activity Assay, Isolation, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Western Blot

Journal: Frontiers in Microbiology

Article Title: Lily polysaccharides alleviate colitis through the microbiota–N8-acetylspermidine–cGAS–STING signaling axis

doi: 10.3389/fmicb.2025.1686902

Figure Lengend Snippet: In vitro verification, lily polysaccharide can inhibit the cGAS-STING pathway and play an anti-inflammatory role. (a) IL-1β mRNA expression. (b) TNF-α mRNA expression level. (c) IL-6 mRNA expression level. (d) ZO-1 mRNA expression level. (e) occludin mRNA expression level. (f) MUC2 mRNA expression level. (g–k) The protein expression levels of cGAS, STING, p-TBK, p-IRF3 were assessed by western blot analysis. Data are presented as the mean ± SEM. Statistically significant differences are indicated; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Subsequently, the samples were washed again with PBST and incubated overnight at 4 °C with a primary antibody against MUC2 (Rabbit polyclonal IgG, Proteintech, Catalog No. 27675-1-AP, unconjugated; 1:200 dilution).

Techniques: In Vitro, Expressing, Western Blot

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Specific activation of hypoxia-inducible factor-2α by propionate metabolism via a β-oxidation-like pathway stimulates MUC2 production in intestinal goblet cells.

doi: 10.1016/j.biopha.2022.113672

Figure Lengend Snippet: Fig. 1. Propionate-mediated cytotoxicity and MUC2 production in LS 174 T cells. LS 174 T cells were treated with various concentrations of propionate at 37 ◦C for 24 h, and cytotoxicity was measured by MTT assay (A). Expression of MUC2 (B) was measured by quantitative real-time PCR. Total and intracellular MUC2 protein were detected by ELISA (C) and immunofluorescence (D) and quantified (E), respectively. Data are expressed as the mean ± SD of three independent experiments performed in triplicate. Student’s t-test was used to determine the significance of the differences.

Article Snippet: The Human MUC2 ELISA kit (E-EL-H0632, Elabscience, Houston, TX, USA) was used to measure total MUC2 levels.

Techniques: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Specific activation of hypoxia-inducible factor-2α by propionate metabolism via a β-oxidation-like pathway stimulates MUC2 production in intestinal goblet cells.

doi: 10.1016/j.biopha.2022.113672

Figure Lengend Snippet: Fig. 3. Hypoxic response of LS 174 T cells to propionate treatment. Cells were treated with propionate (1–4 mM) for 24 h, and changes in HIF expression were investigated. (A) The transcriptional level of the α subunits of HIFs at 24 h. (B) The results of a time-course assay comparing MUC2 levels with those of the HIFs using qPCR. (C and D) Cell lysates were fractionated into cytosolic and nuclear fractions, and activated HIF-α levels were detected by western blot analysis (C) and quantified (D). Data are expressed as the mean ± SD of three independent experiments performed in triplicate. Student’s t-test was used to determine the significance of the differences: ns, not significant. NC, negative control; Pro, propionate-treated. The full-length blots of (C) are shown in Fig. S4.

Article Snippet: The Human MUC2 ELISA kit (E-EL-H0632, Elabscience, Houston, TX, USA) was used to measure total MUC2 levels.

Techniques: Expressing, Western Blot, Negative Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Specific activation of hypoxia-inducible factor-2α by propionate metabolism via a β-oxidation-like pathway stimulates MUC2 production in intestinal goblet cells.

doi: 10.1016/j.biopha.2022.113672

Figure Lengend Snippet: Fig. 4. HIF-2α was activated by propionate-induced mitochondrial respiration. (A) Propionate (4 mM) and oligomycin A (1 μM) were added to LS 174 T cells and incubated for 12 h, and ATPase activity was measured. (B–D) The suppressive effects of oligomycin A on HIF-α were investigated at the transcriptional level (B) and nuclear protein levels were measured by western blotting (C) and quantified (D). (E) Expression level of MUC2 was measured by qPCR. (F) Total MUC2 level was measured by ELISA. (G and H) Intracellular MUC2 (magnification 600 ×; scale bars: 50 µm) was measured immunofluorescence (G) and quantified (H). Linear correlation of MUC2 and HIF-2α expression by propionate (I). Data are expressed as the mean ± SD of three independent experiments performed in triplicate. One- way ANOVA followed by Tukey’s HSD was used to determine significance: ns, not significant. NC, negative control; Pro, propionate-treated. The full-length blots of (C) are shown in Fig. S5.

Article Snippet: The Human MUC2 ELISA kit (E-EL-H0632, Elabscience, Houston, TX, USA) was used to measure total MUC2 levels.

Techniques: Incubation, Activity Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Negative Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Specific activation of hypoxia-inducible factor-2α by propionate metabolism via a β-oxidation-like pathway stimulates MUC2 production in intestinal goblet cells.

doi: 10.1016/j.biopha.2022.113672

Figure Lengend Snippet: Fig. 5. HIF-2α is the primary regulator of MUC2 expression under hypoxia. LS 174 T cells were transfected with mock siRNA or HIF-2α siRNA. (A–C) The effect of 4 mM propionate on the HIF-α subunits was measured at the gene level (A) and nuclear protein levels of HIF-α subunits were measured by western blotting (B) and quantified (C). (D–G) The suppressive effect of HIF-2α siRNA on MUC2 expression was detected at the gene level (D), total protein level of MUC2 was measured by ELISA (E), and intracellular MUC2 protein level (magnification 600 ×; scale bars: 50 µm) was measured by immunofluorescence (F) and quantified (G). Data are expressed as the mean ± SD of three independent experiments performed in triplicate. One-way ANOVA followed by Tukey’s HSD was performed: ns, not significant. NC, negative control; Pro, propionate-treated. The full-length blots of (B) are shown in Fig. S6.

Article Snippet: The Human MUC2 ELISA kit (E-EL-H0632, Elabscience, Houston, TX, USA) was used to measure total MUC2 levels.

Techniques: Expressing, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Negative Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Specific activation of hypoxia-inducible factor-2α by propionate metabolism via a β-oxidation-like pathway stimulates MUC2 production in intestinal goblet cells.

doi: 10.1016/j.biopha.2022.113672

Figure Lengend Snippet: Fig. 7. Propionate metabolism via a β-oxidation-like pathway. LS 174 T cells were transfected with mock siRNA or ACADS siRNA. The cells were then treated with propionate (4 mM) for 24 h. (A) The expression level of ACADS was measured by qPCR. (B) ATPase activity. (C) Expression of the HIF genes was measured by qPCR. (D and E) Nuclear level of HIF proteins were analyzed by western blotting (D) and quantified (E). (F) Expression level of the MUC2 was measured by qPCR. (G) Total MUC2 protein level was measured by ELISA. (H and I) Intracellular MUC2 (magnification 600 ×; scale bars: 50 µm) was measured by immunofluorescence (H) and quantified (I). Data are expressed as the mean ± SD of three independent experiments performed in triplicate. One-way ANOVA followed by Tukey’s HSD was used to determine the significance of differences: ns, not significant. NC, negative control; Pro, propionate-treated; ACADS, short-chain specific acyl-CoA dehydrogenase. The full-length blots of (D) are shown in Fig. S7.

Article Snippet: The Human MUC2 ELISA kit (E-EL-H0632, Elabscience, Houston, TX, USA) was used to measure total MUC2 levels.

Techniques: Transfection, Expressing, Activity Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Negative Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Specific activation of hypoxia-inducible factor-2α by propionate metabolism via a β-oxidation-like pathway stimulates MUC2 production in intestinal goblet cells.

doi: 10.1016/j.biopha.2022.113672

Figure Lengend Snippet: Fig. 8. Hypoxia induced by propionate is not relevant to the carboxylation pathway in LS 174 T cells. LS 174 T cells were transfected with mock-siRNA or MUT siRNA. The cells were then treated with propionate (4 mM) for 24 h. (A) Expression level of MUT was measured by qPCR. (B) ATPase activity. (C) Expression levels of the HIF genes were measured by qPCR. (D and E) The nuclear levels of HIF proteins were analyzed by western blotting (D) and quantified (E). (F) Expression level of the MUC2 gene was measured by qPCR. (G) Total MUC2 level was measured by ELISA. (H and I) Intracellular MUC2 levels were measured by immunofluorescence (magnification 600 ×; scale bars: 50 µm) (H) and quantified (I). Data are expressed as the mean ± SD of three independent experiments performed in triplicate. One- way ANOVA followed by Tukey’s HSD was used to determine the significance of the differences: ns, not significant. NC, negative control; Pro, propionate-treated; MUT, methylmalonyl-CoA mutase. The full-length blots of (D) are shown in Fig. S8.

Article Snippet: The Human MUC2 ELISA kit (E-EL-H0632, Elabscience, Houston, TX, USA) was used to measure total MUC2 levels.

Techniques: Transfection, Expressing, Activity Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Negative Control

Journal: Marine Drugs

Article Title: Chitosan Oligosaccharides Alleviate Colitis by Regulating Intestinal Microbiota and PPARγ/SIRT1-Mediated NF-κB Pathway

doi: 10.3390/md20020096

Figure Lengend Snippet: Oral COS administration ameliorated intestinal mucosal damage and reduced the inflammatory response in DSS-induced colitis mice. ( A ) Images of H&E-stained colon and ileum sections from DSS-treated mice with or without COS administration. The scale bar is 200 μm or 100 μm, as indicated in the respective images. Pathological scores of these tissues were calculated. ( B ) The levels of IL-6, IL-1β and TNF-α in serum were detected by ELISA. ( C ) The level of MUC2 in colonic tissue was detected by ELISA. *, p < 0.05, **, p < 0.01, n = 6.

Article Snippet: Mouse Mucin2 (MUC2) ELISA kit was purchased from CUSABIO (Wuhan, China).

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Journal of Functional Foods

Article Title: Lactobacillus plantarum ZJUIDS04 alleviates DSS-induced colitis via modulating gut microbiota

doi: 10.1016/j.jff.2023.105794

Figure Lengend Snippet: Fig. 4. Effect of L. plantarum ZJUIDS04 on gut-barrier barrier function in DSS-induced colitis mice. (A) Expression of ZO-1, claudin-1, occludin, and in colon tissue evaluated by immunofluorescence staining. Scale bars, 100 μm. (B) Quantitative analysis of ZO-1, claudin-1, and occludin. (C) MUC2. (D) Relative expression of MUC2. All data are expressed as means ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001, vs the DSS group.

Article Snippet: Western blotting was performed as described (Ding et al., 2022) using the following antibodies: MUC2 (Affnity Biosciences, USA), and anti-β-actin (Boster C. Yu et al. Journal of Functional Foods 109 (2023) 105794 Biological Technology, Wuhan, China).

Techniques: Expressing, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: Dietary polyamines promote intestinal adaptation in an experimental model of short bowel syndrome

doi: 10.1038/s41598-024-55258-4

Figure Lengend Snippet: Polyamines enhance mucosal defense factors in rats with massive intestinal resection. (A-B) Fecal and serum secretory IgA was measured using ELISA. n = 5–7/group. (C) IgA in ileum tissue was assessed using western blotting. n = 4–6/group. (D) Fecal mucin was measured using a fluorometric assay. n = 4–6/group. (E) Representative images of ileal villus showing immunostaining by Muc2 (original magnification, ×400; scale bars = 200 μm). Left graphs show the number of goblet cells per unit villous area and the size of goblet cell secretion granule. (F) The expression of Claudin-3 in the ileum tissue was measured by western blot analysis. Representative images of ileal villus showing immunostaining by Claudin-3 (original magnification, ×200; scale bars = 100 μm). (G) Serum DAO was measured by ELISA. n = 5–7/group. (H) Serum GLP-2 was measured by ELISA. n = 5–7/group. (I) Fecal short-chain fatty acid (SCFA) content was measured using high-performance liquid chromatography. n = 3–6/group. Data are presented as mean ± SD. Results of one-way ANOVA are represented as follows: * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Nonspecific antibody binding was blocked using 15% goat serum for 30 min. For staining of Ki-67, Muc2, and Claudin-3, the slides were then incubated with anti-Ki67 antibody (ab16667; Abcam, Cambridge, UK), anti-Muc2 rabbit monoclonal antibody (M01212-1; Boster Biological Technology, Pleasanton, CA, USA), and Claudin-3 polyclonal antibody (#PA5-32353; Invitrogen, Waltham, MA, USA) at 4°C overnight, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Immunostaining, Expressing, High Performance Liquid Chromatography