combination with e0703  (MedChemExpress)

Journal: mLife

Article Title: The synthetic estradiol analog E0703 enhances Akkermansia muciniphila growth for radiation‐induced intestinal damage repair

doi: 10.1002/mlf2.70071

Figure Lengend Snippet: E0703 enhances survival and intestinal repair following irradiation in mice. (A) Structure of E0703 and schematic diagram of animal intervention. E0703, a structural analog of estrogen, was administered to mice at 5 mg/kg 24 h prior to irradiation. Intestinal tissues and fecal samples were collected on Day 5 (D5) post irradiation. (B) Body weight analysis of different groups of mice 1 week after irradiation. One week after irradiation, body weight in the irradiated (IR) group declined significantly, whereas Amifostine (IRA) and E0703 (IRB) groups demonstrated weight recovery from Day 4 to Day 5. (C) Survival analysis of mice 1 week after irradiation. The survival rate in the IR group decreased markedly within 1‐week post irradiation, while IRA and IRB groups maintained approximately 80% survival. (D) Histopathological results of the small intestine on Day 5 after irradiation. Irradiation disrupted small intestine structure, reduced villus number and length, and decreased the fluorescence intensity of the mucin protein Muc2. Both Amifostine and E0703 preserved villus structure and Muc2 expression, with E0703 showing a more pronounced effect. Scale bars: 100 μm and 400 μm. (E, F) Quantitative analysis of small intestinal pathology (E) and Muc2 expression (F). A trend toward improved villus length with E0703 is shown, though this is not statistically significant. E0703 significantly enhanced Muc2 expression compared with the IR group ( p < 0.05), showing greater efficacy than Amifostine. (G) Sobs index analysis of intestinal microbiota in mice on Day 5 after irradiation. Radiation significantly reduced gut microbiota richness, with no significant improvement in drug‐treated groups. (H) PCA analysis of intestinal microbiota in mice on Day 5 after irradiation. Beta diversity analysis demonstrated distinct clustering among groups, with IRB and CON groups showing the closest distance. (I) Phylum‐level composition of intestinal microbiota in mice on Day 5 after irradiation. Radiation caused significant changes in gut microbiota composition, notably an increased abundance of Verrucomicrobiota in the IRB group. (J) Species‐level composition of intestinal microbiota in mice on Day 5 after irradiation. At the species level, Akkermansia muciniphila (AKK) abundance was significantly higher in the IRB group. (K) Comparison of intestinal AKK abundance in mice on Day 5 after irradiation. E0703 significantly increased AKK in irradiated mice ( p < 0.05). (L) LEfSe analysis of intestinal microbiota in mice on Day 5 after irradiation. Significant microbial groups was identified, with higher LDA scores indicating a greater effect of species abundance. CON, nonirradiated mice; LDA, linear discriminant analysis; LEfSe, LDA effect size; OTU, operational taxonomic unit; PCA, principal component analysis. Group differences were evaluated using one‐way analysis of variance (ANOVA). ns, not significant. * p < 0.05; ** p < 0.01; and *** p < 0.001.

Article Snippet: An additional group was treated with Esr2 antagonist 4‐[2‐Phenyl‐5,7‐bis(trifluoromethyl)pyrazolo[1,5‐a]pyrimidin‐3‐yl]phenol (PHTPP) (1 μM, MCE, HY‐103456) in combination with E0703 (60 ng/ml).

Techniques: Irradiation, Fluorescence, Expressing, Comparison

Journal: mLife

Article Title: The synthetic estradiol analog E0703 enhances Akkermansia muciniphila growth for radiation‐induced intestinal damage repair

doi: 10.1002/mlf2.70071

Figure Lengend Snippet: E0703 increases AKK abundance in the mouse intestine. (A) Schematic diagram of fecal microbiota sampling after E0703 intervention under nonirradiated conditions. Fecal samples from E0703‐treated normal mice were collected on Day 1, 5, and 10 for 16S rDNA sequencing. (B) Sobs index analysis of intestinal microbiota at different time points after E0703 intervention. A nonsignificant trend toward increased richness following E0703 treatment is shown. (C) PCA analysis of intestinal microbiota at different time points after E0703 intervention. E0703 altered gut microbiota structure over time. (D) Phylum‐level composition of intestinal microbiota at different time points after E0703 intervention. The relative abundance of Verrucomicrobiota increased significantly on Day 1, 5, and 10. (E) Species‐level composition of intestinal microbiota at different time points after E0703 intervention. E0703 significantly modified species‐level microbiota composition over time, increasing beneficial bacteria such as AKK, Bacteroides acidifaciens , Faecalibaculum rodentium , and Bifidobacterium pseudolongum . (F) Comparative analysis of the abundance of four probiotics at different time points after E0703 intervention. Single‐dose E0703 administration more effectively increased the abundance of these probiotics on Day 5 than on Day 10. Group differences were evaluated using one‐way analysis of variance (ANOVA). * p < 0.05; ** p < 0.01.

Article Snippet: An additional group was treated with Esr2 antagonist 4‐[2‐Phenyl‐5,7‐bis(trifluoromethyl)pyrazolo[1,5‐a]pyrimidin‐3‐yl]phenol (PHTPP) (1 μM, MCE, HY‐103456) in combination with E0703 (60 ng/ml).

Techniques: Sampling, Sequencing, Modification, Bacteria, Probiotics

Journal: mLife

Article Title: The synthetic estradiol analog E0703 enhances Akkermansia muciniphila growth for radiation‐induced intestinal damage repair

doi: 10.1002/mlf2.70071

Figure Lengend Snippet: AKK supplementation potentiates E0703‐mediated radiation resistance and intestinal protection. (A) Schematic diagram of AKK intervention in irradiated mice. Mice received AKK by gavage every 3 days for 10 days prior to irradiation. Intestinal tissues and fecal samples were collected on Day 5 post irradiation. (B) Body weight analysis of mice 1 week after irradiation under different drug interventions. AKK gavage did not affect body weight before irradiation but improved recovery in irradiated mice on Day 4 and 5. (C) Survival analysis of mice 1 week after irradiation under different drug interventions. AKK alone enhanced survival after irradiation, while its combination with E0703 maintained 90% survival. (D, E) Colon length and its quantification on Day 5 after irradiation under different drug interventions. Irradiation reduced rectal length, but AKK or E0703, alone or combined, improved rectal length ( p < 0.05). (F–K) Histopathology of the small intestine and quantitative analysis on Day 5 after irradiation under different drug interventions. Histological analysis showed that AKK, E0703, and their combination improved villus length, PAS‐positive area, and fluorescence area proportions of Muc2, Lgr5, and Ki67, with the combination group showing the best effect. Scale bar: 100 μm (F). (L, M) Representative images and quantification of intestinal organoid intervention with fecal supernatants. Fecal supernatants from treated mice promoted intestinal organoid growth, with the combination group demonstrating the most pronounced effect. IRAK, AKK‐treated; IRBAK, AKK‐, and E0703‐treated before irradiation. F, fecal supernatant; PAS, Periodic acid‐Schiff. Group differences were evaluated using one‐way analysis of variance (ANOVA). * p < 0.05; and ** p < 0.01. Scale bar: 100 μm (L).

Article Snippet: An additional group was treated with Esr2 antagonist 4‐[2‐Phenyl‐5,7‐bis(trifluoromethyl)pyrazolo[1,5‐a]pyrimidin‐3‐yl]phenol (PHTPP) (1 μM, MCE, HY‐103456) in combination with E0703 (60 ng/ml).

Techniques: Irradiation, Histopathology, Fluorescence

Journal: mLife

Article Title: The synthetic estradiol analog E0703 enhances Akkermansia muciniphila growth for radiation‐induced intestinal damage repair

doi: 10.1002/mlf2.70071

Figure Lengend Snippet: E0703 modulates the transcriptome and metabolome of AKK. (A) OD 600 analysis of AKK after 48 h E0703 intervention in vitro . Optical density (OD) measurements showed no significant effect of E0703 on AKK growth. (B) OD 600 analysis of AKK from 12 to 96 h after Muc2 intervention in vitro . Recombinant human Muc2 protein enhanced AKK growth in a dose‐ and time‐dependent manner. (C) Schematic diagram of supernatant metabolomics and pellet transcriptomics after a 48 h E0703 intervention in AKK in vitro . Blank, normal medium; CON, AKK without intervention; DMSO, AKK with dimethyl sulfoxide; and Drug, AKK with E0703 in DMSO. (D) Volcano plot comparing gene expression between E0703 and DMSO groups ( p < 0.05, log 2 (fold change) >0.5). (E) Heatmap of differentially expressed genes clustered by M‐fuzz, with KEGG enrichment shown. (F) Heatmap of significantly different metabolites, highlighting those with higher abundance in the E0703 group. (G) Comparison of GlcN‐6‐P levels in mice feces across groups. (H, I) Representative images and quantitative analysis of intestinal organoids treated with GlcN‐6‐P. GlcN‐6‐P intervention improved the growth of irradiated intestinal organoids ( p < 0.05). Group differences were evaluated using one‐way analysis of variance (ANOVA). ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001. Scale bar: 200 μm (H).

Article Snippet: An additional group was treated with Esr2 antagonist 4‐[2‐Phenyl‐5,7‐bis(trifluoromethyl)pyrazolo[1,5‐a]pyrimidin‐3‐yl]phenol (PHTPP) (1 μM, MCE, HY‐103456) in combination with E0703 (60 ng/ml).

Techniques: In Vitro, Recombinant, Transcriptomics, Gene Expression, Comparison, Irradiation

Journal: mLife

Article Title: The synthetic estradiol analog E0703 enhances Akkermansia muciniphila growth for radiation‐induced intestinal damage repair

doi: 10.1002/mlf2.70071

Figure Lengend Snippet: Single‐cell RNA sequencing (scRNA‐seq) reveals the protective effect of E0703 in radiation‐induced intestinal injury. (A) Schematic of the scRNA‐seq experimental design. (B) Single‐cell atlas of mouse intestines following irradiation and E0703 treatment. (C) Dot plot of normalized mean expression of canonical marker genes across clusters, with dot size indicating the proportion of expression. Average expression denotes the mean expression level of a gene across diverse cell populations, with darker colors indicating higher expression levels. Expressed (%) illustrates the percentage of cells within a particular cell population that expresses the specified gene. (D) Alluvial chart showing cell type proportions in control, IR, and E0703‐treated groups. (E) Changes in cell proportions 5 days post irradiation and after E0703 treatment. ΔCell fraction (%) means the difference in cell fraction between IR_Drug and IR groups. (F) Dot plot of marker gene expression across clusters and conditions, with dot size indicating expression proportion.

Article Snippet: An additional group was treated with Esr2 antagonist 4‐[2‐Phenyl‐5,7‐bis(trifluoromethyl)pyrazolo[1,5‐a]pyrimidin‐3‐yl]phenol (PHTPP) (1 μM, MCE, HY‐103456) in combination with E0703 (60 ng/ml).

Techniques: Single Cell, RNA Sequencing, Irradiation, Expressing, Marker, Control, Gene Expression

Journal: mLife

Article Title: The synthetic estradiol analog E0703 enhances Akkermansia muciniphila growth for radiation‐induced intestinal damage repair

doi: 10.1002/mlf2.70071

Figure Lengend Snippet: E0703 enhances oxidative phosphorylation, increasing intestinal stem and goblet cell ratios after irradiation. (A) Heatmap illustrating normalized mean expression of marker genes in stem and goblet cells. (B, C) Dot plots of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment in stem and goblet cells across conditions. Darker colors indicate smaller p ‐values and larger dots indicate more genes. (D) Genes commonly enriched in the oxidative phosphorylation pathway in both cell types. (E) Gene expression comparison in the oxidative phosphorylation pathway across groups. (F–J) Both Amifostine and E0703 improved mitochondrial complex I–V activity in irradiated intestinal tissue, with E0703 showing greater effect. Group differences were evaluated using one‐way analysis of variance (ANOVA). ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001.

Article Snippet: An additional group was treated with Esr2 antagonist 4‐[2‐Phenyl‐5,7‐bis(trifluoromethyl)pyrazolo[1,5‐a]pyrimidin‐3‐yl]phenol (PHTPP) (1 μM, MCE, HY‐103456) in combination with E0703 (60 ng/ml).

Techniques: Phospho-proteomics, Irradiation, Expressing, Marker, Gene Expression, Comparison, Activity Assay