e2 mce hy b0141  (MedChemExpress)

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Activation of the itaconate metabolic pathway in a mouse model of endometriosis (A) Schematic diagram of the mouse endometriosis model. (B) Representative intra-abdominal images. Top: normal uterus from control mice. Middle and bottom: recipient mice with endometriotic lesions (red circles); lower panels show magnified views. (C) Bubble plot of quantitative enrichment analysis showing altered metabolic pathways among ectopic lesions, eutopic endometrium, and normal endometrium in mice. (D) Heat map of selected metabolites across normal, eutopic, and ectopic tissues from the mouse endometriosis model. Row-wise z-scored intensities (no log transform); colors reflect relative abundance per metabolite, centered at 0 (≈[−1, 1]). (E) Quantification of itaconate in ectopic lesions (n = 10), eutopic endometrium (n = 5), and normal endometrium (n = 5). (F, G) Western blot analysis (F) and quantification (G) of ACOD1 protein in lesion tissues from endometriosis (EM) and control mice (n = 3/group). (H, I) Western blot analysis (H) and quantification (I) of ACOD1 in stromal cells from normal (nor-ESC), eutopic (eu-ESC), and ectopic (ec-ESC) endometrium (n = 3/group). (J) qRT-PCR analysis of Acod1 mRNA in nor-ESC, eu-ESC, and ec-ESC (n = 3/group); LPS-stimulated macrophages from control mice serve as positive control. (Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ns: not significant.) E2, estradiol benzoate; EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Activation Assay, Control, Western Blot, Quantitative RT-PCR, Positive Control

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Pathway Intervention Alters Lesion Burden and Fibrosis in Endometriosis Mice (A)Schematic of IRG1-IN-1 (0.5 mg/kg) or PBS administration in the mouse endometriosis (EM) model. (B) Representative images of endometriotic lesions in PBS- and IRG1-IN-1-treated mice. (C) Gross morphology of excised lesions from each group. (D) Quantification of lesion weight (n = 6/group). (E) Schematic of the experimental design for si-Irg1-LNP administration in EM mice. (F, G) Representative images (F) and gross morphology (G) of lesions in si-ctrl-LNP and si-Irg1-LNP groups. (H) Quantification of lesion weight (n = 6/group). (I) Western blot analysis of ACOD1 expression in lesions from si-ctrl-LNP and si-Irg1-LNP groups.(J) Schematic of the experimental design for 4-OI intervention in EM mice. (K) Representative images of endometriotic lesions after treatment with PBS or 4-OI (red arrows). (L) Gross morphology of lesions from each group. (M) Quantification of lesion weight (n = 6/group). (N) Masson's trichrome staining of lesions with quantification of fibrotic area (n = 6/group). (O) Measurement of itaconate content in peritoneal macrophages from EM and non-EM mice (n = 8/group). (P) Western blot analysis of ACOD1 in peritoneal macrophages from EM and control mice E2, estradiol benzoate; EM, endometriosis; LNP, lipid nanoparticle; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; PBS, phosphate-buffered saline; si- Irg1 , small interfering RNA targeting Irg1.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Western Blot, Expressing, Staining, Control, Saline, Small Interfering RNA

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate-mediated modulation of macrophage activity alters endometriotic lesion p rogression (A) mRNA expression of IL1B , IL6 , TNFA , and iNOS in peritoneal macrophages from endometriosis (EM) and non-EM patients, as well as non-EM macrophages co-cultured with either nor-ESC or ectopic ec-ESC for 12 h (n = 3/group).(B) Migration of si- Irg1 or NC-treated ectopic ESCs was assessed after co-culture with PBMCs for 48 h. Quantification of migrated cells in five random fields per group are shown (n = 3/group). (C-D) PKH67-labeled human ectopic ESCs pretreated with si- Irg1 or NC were co-cultured with PBMCs for 8 h. Phagocytosis of ESCs by macrophages was analyzed by flow cytometry with representative gating (C), PKH67 signal and quantification of PKH67-positive macrophages (D) (n = 6/group).(E,G) Flow cytometry analysis and quantification of iNOS + peritoneal macrophages from mouse model in E (E), A (G). (n = 6/group). (F,H) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in peritoneal macrophages from mouse model in E (E), A (G) (n = 6/group).(I)Quantification of itaconate in endometriotic lesion tissue by LC–MS from mice treated with IRG1-IN-1 or vehicle. (n = 3/group).(J) Flow cytometry analysis and quantification of iNOS + peritoneal macrophages from mouse model in J (n = 6/group).(K,L) Migration of mESCs induced by peritoneal macrophages from PBS- or 4-OI-treated mice was assessed by transwell assay; representative images and quantification of migrated cells are shown (n = 5/group). (M) Flow cytometry analysis and quantification of phagocytosis of PKH67-labeled mESCs by peritoneal macrophages (n = 6/group).(N) Schematic of the experimental design for clodronate liposome-mediated macrophage depletion and 4-OI intervention in EM mice. (O, P) Representative images (O) and gross morphology (P) of endometriotic lesions in control, clodronate, PBS, and 4-OI groups. (Q) Quantification of lesion weight in mice treated with clodronate liposomes or control liposomes, with or without 4-OI (n = 6/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) E2, estradiol benzoate; EM, endometriosis; LNP, lipid nanoparticle; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; Clod, clodronate liposome; PBS, phosphate-buffered saline; si- Irg1 , small interfering RNA targeting Irg1.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Activity Assay, Expressing, Cell Culture, Migration, Co-Culture Assay, Labeling, Flow Cytometry, Liquid Chromatography with Mass Spectroscopy, Transwell Assay, Control, Liposomes, Saline, Small Interfering RNA

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Inhibition of NRF2 Signaling Reverses the Anti-inflammatory and Anti-endometriotic Effects of Itaconate in Macrophages and a Mouse Model of End ometriosis (A,B) Western blot analysis (A) and quantification (B) of NRF2 protein levels in bone marrow-derived macrophages (BMDMs) pretreated with LPS (100 ng/mL), 4-octyl itaconate (4-OI, 250 μM), or both for 12 h (n = 3/group). (C,D) Western blot analysis (C) and quantification (D) of NRF2 in BMDMs treated with LPS, 4-OI, and the NRF2 inhibitor ML385 (2.5 μM) for 12 h (n = 3/group). (E) Flow cytometry analysis and quantification of iNOS + BMDMs after indicated treatments (n = 3/group). (F) mRNA expression of pro-inflammatory genes ( Il1b , Il6 , Nos2 , Tnf ) in BMDMs under different conditions (n = 3/group). (G) Flow cytometry analysis and quantification of iNOS + BMDMs following NRF2 knockdown (si Nrf2 ) with or without 4-OI, compared to negative control (NC) (n = 3/group).(H) Schematic of the experimental design for ML385 administration in a mouse model of endometriosis. (I) Representative images of endometriotic lesions in PBS- and ML385-treated mice (lesions marked by red circles). (J) Gross morphology of lesions in both groups(n = 6/group). (K) Quantification of lesion weight (n = 6/group). (L) Flow cytometry analysis and quantification of iNOS + peritoneal macrophages from EM mice treated with PBS or ML385 (n = 6/group). (M) mRNA expression of Il1b , Il6 , Nos2 , and Tnf in peritoneal macrophages from each group (n = 6/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) E2, estradiol benzoate; EM, endometriosis; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; ML385, NRF2 inhibitor.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Inhibition, Western Blot, Derivative Assay, Flow Cytometry, Expressing, Knockdown, Negative Control

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Suppresses NOX2-Derived ROS to Regulate Macrophage Function and Lesion Progression in End ometriosis (A-B) qPCR analysis of NOX2 mRNA in peritoneal macrophages (PMs) from human (A) and mouse (B) NC and EM groups (n = 3/group). (C-D) Western blot and quantification of NOX2 protein in PBMC-derived macrophages treated with LPS or LPS + 4-OI (n = 5/group). (E) NOX2 enzyme activity in BMDMs (n = 4/group). (F, G) Flow cytometry and quantification of ROS production in BMDMs after LPS or LPS + 4-OI (n = 3/group). (H, I) Flow cytometry and quantification of iNOS + BMDMs after LPS, LPS + 4-OI, or LPS + 4-OI + DPI treatment (n = 4/group). (J) Intracellular Ca 2+ dynamics in BMDMs measured by Fluo-4 after LPS, 4-OI, or DPI treatment. (K) Schematic of 4-OI and DPI intervention in the mouse endometriosis model. (L, M) Representative images (L) and gross morphology (M) of endometriotic lesions after PBS, 4-OI, or DPI treatment. (N) Quantification of lesion weight (n = 6/group). (O, P) Flow cytometry and quantification of iNOS + peritoneal macrophages (percentage and MFI) in peritoneal lavage (n = 6/group). (Q) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in peritoneal macrophages after treatments (n = 6/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001, ns: not significant.) E2, estradiol benzoate; EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; DPI, NOX2 inhibitor.

Article Snippet: Estrous cycles were synchronized with subcutaneous estradiol benzoate (E2) (3 μg/mouse) (MCE; Nanjing, China).

Techniques: Derivative Assay, Western Blot, Activity Assay, Flow Cytometry, Control