Journal: Aging Cell

Article Title: Aging Triggers an Intestinal Energy Crisis and HDL3 Deficiency Disrupting Gut–Liver Axis Homeostasis

doi: 10.1111/acel.70445

Figure Lengend Snippet: Mitochondrial dysfunction‐mediated ATP deficiency suppresses HDL3 synthesis in aging intestinal cells. (a) Representative images (scale bar: 5 μm) of intestinal cell microstructure measured by TEM, n = 18 images from n = 3 independent experiments; (b) ileum ATP levels, n = 5; (c) representative images (scale bar: 5 μm) of IME microstructure measured by TEM, n = 18 images from n = 3 independent experiments; (d) IME ATP levels, n = 5; (e) glycolysis assay measured as cytoplasmic acidification, the fluorescence signal was enhanced with the increase of acidification degree, n = 4; (f) oxygen consumption, as mitochondrial respiration depletes the oxygen within the assay medium, quenching of the fluorescent dye is reduced, and the fluorescence signal increases proportionately, n = 4; (g, h) OXPHOS protein expression levels in the ileum, n = 3; and (i) exogenous ATPγS‐AM (50 μM) partially restored HDL3 synthesis in senescent IME cells, whereas native ATP (50 μM) had no significant effect, n = 5. Data are express as the mean ± SEM. * p < 0.05, ** p < 0.01. D‐Gal: D‐galactose; NC, normal control.

Article Snippet: In the normal control groups (10 replicates), the medium was replaced with complete DMEM, whereas in the model control groups (10 replicates), the medium was replaced with D‐galactose (200 mM, dissolved in complete DMEM) and cultured for 24 h. After successful establishment of the aging model, the medium was discarded, and the model control group (D‐Gal) (replaced with complete DMEM containing ApoA1 10 μg/mL), ATP intervention group (D‐Gal‐ATP) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATP 50 μM, ATPγS‐AM intervention group (D‐Gal‐ATPγS‐AM) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATPγS‐AM 50 μM, NMN intervention group (D‐Gal‐NMN) was replaced with complete DMEM containing ApoA1 10 μg/mL and NMN 5 μM, the agonist CS‐6253 (MedChem Express, Shanghai, China) intervention group (D‐GAL‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL and CS‐6253 1 μM; and the NMN and CS‐6253 synergistic group (D‐Gal‐NMN‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL, NMN 5 μM, and CS‐6253 1 μM.

Techniques: Fluorescence, Expressing, Control

Journal: Aging Cell

Article Title: Aging Triggers an Intestinal Energy Crisis and HDL3 Deficiency Disrupting Gut–Liver Axis Homeostasis

doi: 10.1111/acel.70445

Figure Lengend Snippet: ABCA1 downregulation limits HDL3 synthesis in aging. (a) Relative mRNA expression of ABCA1 , ApoA1 , LPL , and ANGPTL3 in ileum, n = 5; (b, c) representative images (scale bar: 50 μm) and quantitative analysis of ABCA1, ApoA1, LPL, and ANGPTL3, measured by IHC staining, n = 3; and (d) activation of ABCA1 expression combined with ATPγS‐AM supplementation enhances cellular HDL3 synthesis capacity n = 5. Data are express as the mean ± SEM. ** p < 0.01. ABCA1, ATP‐binding cassette transporter 1; ANGPTL3, angiopoietin‐like3; CS‐6253, ABCA1 activators; D‐Gal, D‐galactose; HDL3, high‐density lipoprotein 3; IHC, immunohistochemistry; IME, intestinal mucosa epithelial; LPL, lipoprotein lipase; NC, normal control.

Article Snippet: In the normal control groups (10 replicates), the medium was replaced with complete DMEM, whereas in the model control groups (10 replicates), the medium was replaced with D‐galactose (200 mM, dissolved in complete DMEM) and cultured for 24 h. After successful establishment of the aging model, the medium was discarded, and the model control group (D‐Gal) (replaced with complete DMEM containing ApoA1 10 μg/mL), ATP intervention group (D‐Gal‐ATP) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATP 50 μM, ATPγS‐AM intervention group (D‐Gal‐ATPγS‐AM) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATPγS‐AM 50 μM, NMN intervention group (D‐Gal‐NMN) was replaced with complete DMEM containing ApoA1 10 μg/mL and NMN 5 μM, the agonist CS‐6253 (MedChem Express, Shanghai, China) intervention group (D‐GAL‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL and CS‐6253 1 μM; and the NMN and CS‐6253 synergistic group (D‐Gal‐NMN‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL, NMN 5 μM, and CS‐6253 1 μM.

Techniques: Expressing, Immunohistochemistry, Activation Assay, Binding Assay, Control

Journal: Aging Cell

Article Title: Aging Triggers an Intestinal Energy Crisis and HDL3 Deficiency Disrupting Gut–Liver Axis Homeostasis

doi: 10.1111/acel.70445

Figure Lengend Snippet: Aging impairs ABCA1‐mediated cholesterol efflux and reduces HDL3 synthesis. (a, b) Representative images (scale bar: 100 μm) of ABCA1 measured by IF staining in ileum and IME, n = 6 images from n = 3 independent experiments; and (c, d) efficiency of cholesterol efflux to ApoA‐1 and HDL, n = 5. Data are expressed as the mean ± SEM. ** p < 0.01. ABCA1, ATP‐binding cassette transporter 1; CS‐6253, ABCA1 activators; D‐Gal, D‐galactose; IF, immunofluorescence; IME, intestinal mucosa epithelial; NC, normal control.

Article Snippet: In the normal control groups (10 replicates), the medium was replaced with complete DMEM, whereas in the model control groups (10 replicates), the medium was replaced with D‐galactose (200 mM, dissolved in complete DMEM) and cultured for 24 h. After successful establishment of the aging model, the medium was discarded, and the model control group (D‐Gal) (replaced with complete DMEM containing ApoA1 10 μg/mL), ATP intervention group (D‐Gal‐ATP) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATP 50 μM, ATPγS‐AM intervention group (D‐Gal‐ATPγS‐AM) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATPγS‐AM 50 μM, NMN intervention group (D‐Gal‐NMN) was replaced with complete DMEM containing ApoA1 10 μg/mL and NMN 5 μM, the agonist CS‐6253 (MedChem Express, Shanghai, China) intervention group (D‐GAL‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL and CS‐6253 1 μM; and the NMN and CS‐6253 synergistic group (D‐Gal‐NMN‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL, NMN 5 μM, and CS‐6253 1 μM.

Techniques: Staining, Binding Assay, Immunofluorescence, Control

Journal: Aging Cell

Article Title: Aging Triggers an Intestinal Energy Crisis and HDL3 Deficiency Disrupting Gut–Liver Axis Homeostasis

doi: 10.1111/acel.70445

Figure Lengend Snippet: NMN modulates mitochondrial function to boost ATP production in the aging intestine. (a, b) NADH levels and NAD + /NADH ratio in ileum, n = 3; (c) relative telomere length in ileum (T/S), n = 5; (d, e) DAO and D‐LA levels in serum, n = 5; (f) ileum relative mRNA expression of Occludin and Claudin‐1 , n = 6; (g, h) representative images (scale bar: 100 μm) and quantitative analysis of Occludin and Claudin‐1 measured by IF staining, n = 6 images from n = 3 independent experiments; (i, k) representative images (scale bar: 5 μm, scale bar: 1 μm) of ileum and IME cell structure measured by TEM, n = 18 images from n = 3 independent experiments; (j, l) ATP levels in ileum and IME cell, n = 5; (m) glycolysis assay measured as cytoplasmic acidification, the fluorescence signal was enhanced with the increase of acidification degree, n = 4; (n) oxygen consumption, as mitochondrial respiration depletes the oxygen within the assay medium, quenching of the fluorescent dye is reduced, and the fluorescence signal increases proportionately, n = 4; and (o, p) OXPHOS protein expression levels in the ileum, n = 3. Data are express as the mean ± SEM. * p < 0.05, ** p < 0.01. DAO, diamine oxidase; D‐Gal, D‐galactose; D‐LA, D‐lactic acid; IF, immunofluorescence; IME, intestinal mucosa epithelial; NC, normal control.

Article Snippet: In the normal control groups (10 replicates), the medium was replaced with complete DMEM, whereas in the model control groups (10 replicates), the medium was replaced with D‐galactose (200 mM, dissolved in complete DMEM) and cultured for 24 h. After successful establishment of the aging model, the medium was discarded, and the model control group (D‐Gal) (replaced with complete DMEM containing ApoA1 10 μg/mL), ATP intervention group (D‐Gal‐ATP) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATP 50 μM, ATPγS‐AM intervention group (D‐Gal‐ATPγS‐AM) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATPγS‐AM 50 μM, NMN intervention group (D‐Gal‐NMN) was replaced with complete DMEM containing ApoA1 10 μg/mL and NMN 5 μM, the agonist CS‐6253 (MedChem Express, Shanghai, China) intervention group (D‐GAL‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL and CS‐6253 1 μM; and the NMN and CS‐6253 synergistic group (D‐Gal‐NMN‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL, NMN 5 μM, and CS‐6253 1 μM.

Techniques: Expressing, Staining, Fluorescence, Immunofluorescence, Control

Journal: Aging Cell

Article Title: Aging Triggers an Intestinal Energy Crisis and HDL3 Deficiency Disrupting Gut–Liver Axis Homeostasis

doi: 10.1111/acel.70445

Figure Lengend Snippet: NMN enhances intestinal HDL3 synthesis in the aging intestine. (a, b) NMN enhanced HDL3 synthesis capacity in the ileum and IME cells, n = 5; (c–e) NMN increased the relative expression of ABCA1 mRNA and protein in the ileum. n = 3; (f, h) representative images (scale bar: 100 μm) of ABCA1 localization to the cell membrane measured by IF staining, n = 6 images from n = 3 independent experiments; (g) NMN increased the relative expression of ABCA1 mRNA in the IME cells. n = 3; and (i, j) NMN enhanced cholesterol efflux to ApoA‐1 and HDL in aging cells, n = 5. Data are expressed as the mean ± SEM. ** p < 0.01. ABCA1, ATP‐binding cassette transporter 1; D‐Gal, D‐galactose; HDL, high‐density lipoprotein; IF, immunofluorescence; IME, intestinal mucosa epithelial; NC, normal control.

Article Snippet: In the normal control groups (10 replicates), the medium was replaced with complete DMEM, whereas in the model control groups (10 replicates), the medium was replaced with D‐galactose (200 mM, dissolved in complete DMEM) and cultured for 24 h. After successful establishment of the aging model, the medium was discarded, and the model control group (D‐Gal) (replaced with complete DMEM containing ApoA1 10 μg/mL), ATP intervention group (D‐Gal‐ATP) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATP 50 μM, ATPγS‐AM intervention group (D‐Gal‐ATPγS‐AM) was replaced with complete DMEM containing ApoA1 10 μg/mL and ATPγS‐AM 50 μM, NMN intervention group (D‐Gal‐NMN) was replaced with complete DMEM containing ApoA1 10 μg/mL and NMN 5 μM, the agonist CS‐6253 (MedChem Express, Shanghai, China) intervention group (D‐GAL‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL and CS‐6253 1 μM; and the NMN and CS‐6253 synergistic group (D‐Gal‐NMN‐CS‐6253) was replaced with complete DMEM containing ApoA1 10 μg/mL, NMN 5 μM, and CS‐6253 1 μM.

Techniques: Expressing, Membrane, Staining, Binding Assay, Immunofluorescence, Control

Journal: Stem Cell Research & Therapy

Article Title: HMGB1-mediated enhancement of glycolysis activates hepatic stellate cells by inhibiting ferroptosis in alcoholic hepatic fibrosis

doi: 10.1186/s13287-026-04970-1

Figure Lengend Snippet: HMGB1 regulated ferroptosis to affect HSC activation and ECM deposition. A IHC assay was used to observe the expression levels of GPX4 and FTH1 in liver tissues of AHF patients. B – E The levels of Fe 2+ /Fe 3+ , ROS, GSH content and MDA in LX-2 cells treated with alcohol at different concentrations were evaluated by kit detections. ( F – G ) qRT-PCR and WB assays were employed to assess the effect of elevated alcohol concentration on the mRNA and protein levels of NRF2, GPX4, FTH1, SLC7A11 in alcohol-stimulated LX-2 cells. H – J CCK-8, EdU staining and IF assays were respectively utilized to detect the effect of Erastin on the viability, proliferation, activation and ECM deposition of alcohol-induced LX-2 cells. K The effect of Erastin on lipid peroxidation was assessed by flow cytometry using the C11-BODIPY 581/591 probe in ethanol-induced LX-2 cells. L – P The effects of Ferrostatin 1 on si-HMGB1 regulating the levels of lipid peroxidation, Fe 2+ /Fe 3+ , ROS, GSH content and MDA in alcohol-treated LX-2 cells were measured by kit detections. Q – R qRT-PCR and WB assays were used to evaluate the effects of Ferrostatin 1 on si-HMGB1 regulating the mRNA and protein levels of NRF2, GPX4, FTH1 and SLC7A11 in alcohol-stimulated LX-2 cells. S – T The effect of Ferrostatin 1 on si-HMGB1 regulating the expression of GPX4, FTH1 and HMGB1 in LX-2 cells treated with alcohol was observed by double-labeled IF assay. * p ˂ 0.05, ** p ˂ 0.01, *** p ˂ 0.001 vs. 0 µg/mL Ethanol/ Ethanol/ Ethanol + si-NC/ Ethanol + si-HMGB1

Article Snippet: Mice in the Model + sh-HMGB1 + Ferrostatin-1 group received an intraperitoneal injection of Ferrostatin-1 (1 mg/kg/d) (HY-100579, MedChemExpress, New Jersey, USA) from weeks 6 to 8, while otherwise following the same modeling protocol as described above.

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Concentration Assay, CCK-8 Assay, Staining, Flow Cytometry, Labeling

Journal: Stem Cell Research & Therapy

Article Title: HMGB1-mediated enhancement of glycolysis activates hepatic stellate cells by inhibiting ferroptosis in alcoholic hepatic fibrosis

doi: 10.1186/s13287-026-04970-1

Figure Lengend Snippet: sh-Hmgb1 modulated glycolytic metabolism, ferroptosis, and liver fibrosis in AHF mice. A – B H&E staining and Masson staining were used to observe the effect of sh-Hmgb1 on hepatic fibrosis in AHF mice. C – D The effects of sh-Hmgb1 on Hmgb1‌ mRNA and protein levels in liver tissue of AHF mice were assessed by qRT-PCR and WB assays. E – H Kit and IHC assays were utilized to evaluate the effect of sh-Hmgb1 on the markers related to glycolysis, ferroptosis and HSC activation. * p ˂ 0.05, ** p ˂ 0.01, *** p ˂ 0.001 vs. Normal/ Model + AAV-sh-NC

Article Snippet: Mice in the Model + sh-HMGB1 + Ferrostatin-1 group received an intraperitoneal injection of Ferrostatin-1 (1 mg/kg/d) (HY-100579, MedChemExpress, New Jersey, USA) from weeks 6 to 8, while otherwise following the same modeling protocol as described above.

Techniques: Staining, Quantitative RT-PCR, Activation Assay

Journal: Stem Cell Research & Therapy

Article Title: HMGB1-mediated enhancement of glycolysis activates hepatic stellate cells by inhibiting ferroptosis in alcoholic hepatic fibrosis

doi: 10.1186/s13287-026-04970-1

Figure Lengend Snippet: Hmgb1 influenced liver fibrosis in AHF mice by regulating ferroptosis. A – B H&E staining and Masson staining were utilized to observe the effect of Ferrostatin-1 on sh-Hmgb1 mediating liver fibrosis in AHF mice. C – E The effects of Ferrostatin-1 on sh-Hmgb1 regulating the levels of ROS, GSH content and MDA in liver tissues of AHF mice were assessed by kit detections. F – G IHC assay was employed to measure the effect of Ferrostatin-1 on sh-Hmgb1 regulating the markers related to ferroptosis and HSC activation. * p ˂ 0.05, ** p ˂ 0.01, *** p ˂ 0.001 vs. Model + AAV-sh-NC/ Model + AAV-sh-Hmgb1

Article Snippet: Mice in the Model + sh-HMGB1 + Ferrostatin-1 group received an intraperitoneal injection of Ferrostatin-1 (1 mg/kg/d) (HY-100579, MedChemExpress, New Jersey, USA) from weeks 6 to 8, while otherwise following the same modeling protocol as described above.

Techniques: Staining, Activation Assay

Journal: Bioactive Materials

Article Title: Sustainability of functional hair follicle activity: Impact of spatially anchored multifunctional tetrahedral framework nucleic acids

doi: 10.1016/j.bioactmat.2025.11.006

Figure Lengend Snippet: Establishment of AGA model and treatments. (A) Schematic diagram of the establishment of the AGA model and treatments. (B) Immunofluorescence staining, section scanning, and whole-mount imaging of clarified tissues demonstrating the transdermal efficacy of TQC and tFNAs. Blue: DAPI; Red: Cy5-TQC/Cy5-tFNAs. (C) Time-course observations of hair growth of samples at 1, 4, 7, 10, 14, 15, 18, 21, 24, and 28 days (n = 5). (D) Whole-mount imaging and magnified views of treated samples shot with a Light Microscopy: Control, AGA, Minoxidil, Que, tFNAs, and TQC (n = 5). (D) (E) Longitudinal-sectional and cross-sectional HE staining of treatment groups: Control, AGA, Minoxidil, tFNAs, Que, and TQC. Scale bars: 3 mm (overview), 300 μm/500 μm (magnified regions). Black triangle: Sebaceous gland; black arrowheads: Hair shaft; yellow arrowheads: Hair bulb; green arrowheads: Hair papilla; blue arrowheads: Melanocyte (n = 5). (F) Quantitative image analysis of whole mount hair follicles density (number/view) in figure D. Data are expressed as mean ± SD (n = 5). Statistical significance was determined by one-way ANOVA with Tukey's post hoc test. (G) Quantitative image analysis of hair follicle density (number/mm) in cross-sectional HE staining (figure E). Data are expressed as mean ± SD (n = 5). Statistical significance was determined by one-way ANOVA with Tukey's post hoc test. (H) Ratio of anagen to telogen follicles in longitudinal-sectional HE staining (figure E). Data represent mean ± SD (n = 5) and are analyzed using one-way ANOVA with Tukey's test. The following symbols denote differences compared to each group: (∗) control, (#) AGA, (&) AGA + Minoxidil, (@) AGA + tFNAs, ($) AGA + Que. Significance levels are: ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After 14 days’ model establishment, the mice were assigned to the following groups: AGA model group, Minoxidil ( U10858 , MCE, USA, 5 %)-treated group, Que (20 μM)-treated group, tFNAs (250 nM)-treated group, and TQC (tFNAs/Que: 250 nM/20 μM)-treated group.

Techniques: Immunofluorescence, Staining, Imaging, Light Microscopy, Control

Journal: Bioactive Materials

Article Title: Sustainability of functional hair follicle activity: Impact of spatially anchored multifunctional tetrahedral framework nucleic acids

doi: 10.1016/j.bioactmat.2025.11.006

Figure Lengend Snippet: The role of TQC in mediating Wnt/β-catenin signaling and hair follicle regeneration in AGA mice. (A) Western blot analysis of β-catenin, GSK3β, CK-1α, and APC protein expression in different treatment groups. GAPDH was used as an internal control (n = 5). (B) Quantitative analysis of protein expression levels relative to the control group. Data are expressed as mean ± SD (n = 5). Statistical significance was determined by one-way ANOVA with Tukey's post hoc test. (C) Immunohistochemical staining of GSK3β, CK-1α, and APC in skin tissue sections. Magnified images show representative areas of staining. Scale bars: 3 mm (original magnification) and 500 μm (magnified), (n = 5). (D) Immunofluorescent staining of Ki67 (green) and KRT14 (red) in skin sections. DAPI (blue) was used for nuclear staining. Merged images show the co-localization of Ki67 and KRT14. Scale bars: 2 mm (original magnification) and 200 μm (magnified). (E) Quantification of β-catenin, GSK3β, and CK-1α protein expression on immunofluorescent staining sections calculated by ImageJ (n = 5). Data are expressed as mean ± SD (n = 5). Statistical significance was determined by one-way ANOVA with Tukey's post hoc test. (F) Quantification of Ki67 and KRT14 positive area. Data are expressed as mean ± SD (n = 5). Statistical significance was determined by one-way ANOVA with Tukey's post hoc test. The following symbols denote significant differences compared to each group: (∗) control, (#) AGA, (&) AGA + Minoxidil, (@) AGA + tFNAs, ($) AGA + Que. Significance levels are: ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After 14 days’ model establishment, the mice were assigned to the following groups: AGA model group, Minoxidil ( U10858 , MCE, USA, 5 %)-treated group, Que (20 μM)-treated group, tFNAs (250 nM)-treated group, and TQC (tFNAs/Que: 250 nM/20 μM)-treated group.

Techniques: Western Blot, Expressing, Control, Immunohistochemical staining, Staining