Journal: Acta Pharmaceutica Sinica. B

Article Title: Palmitoylation of Tfr1 enhances platelet ferroptosis and liver injury in heat stroke

doi: 10.1016/j.apsb.2025.10.027

Figure Lengend Snippet: Platelets participate in liver injury in the HS murine model. (A) Thermoregulatory graphical description of heat stress profile aspects within 28 h in mice (shown as mean only, total n = 25). (B) Platelet counts along the time axis at sham heat rest (SHR) and 0, 6, 24 h post-HS ( n = 6 mice per group). (C) Cell cytotoxicity of mouse platelets was measured by LDH activity under heat stress in vitro ( n = 6 mice per group). (D–F) Plasma ALT (D) and AST (E) and IL-1 β (F) levels in SHR mice and mice at 0, 6, 24 h post-HS ( n = 6 mice per group). (G–I) Correlation between platelet counts and Tc max at HS onset ( n = 16), ALT ( n = 24), and IL-1 β ( n = 24) levels. (J–L) Blood platelet counts (J), plasma ALT levels (K), and representative HE staining images of liver tissue (L) were assessed in SHR mice and mice at 0, 6, 24 h post-HS, with or without pretreatment with the platelet-depleting antibody α CD42b or the platelet stimulator thrombopoietin (TPO) ( n = 6 mice per group, scale bars: 100 μm). Mean ± SEM are shown, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: Coagulative factors, interleukin-1 β (IL-1 β ) and lipid peroxidation metabolites were measured by enzyme-linked immunosorbent assay in accordance with the manufacturer's instructions, respectively: IL-1 β (CSB-E08054m/E08053h, CUSABIO, Wuhan, China), IL-6 (CSB-E04639m, CUSABIO), IL-10 (CSB-E04594m-IS, CUSABIO), TNF- α (CSB-E04741m, CUSABIO) and 4-hydroxynonenal (4-HNE, CSB-E13412m/E16214h, CUSABIO).

Techniques: Activity Assay, In Vitro, Clinical Proteomics, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: Palmitoylation of Tfr1 enhances platelet ferroptosis and liver injury in heat stroke

doi: 10.1016/j.apsb.2025.10.027

Figure Lengend Snippet: Platelet-derived IL-1 β contributes to liver injury in the HS murine model. (A) GO (BP) analysis of the physiological and pathological states of platelets in SHR mice and mice at 0 h post-HS (HS0). (B) Representative images of immunofluorescence in liver tissue from SHR and HS mice ( n = 3 independent experiments, scale bars, 50 μm). DAPI (blue), CD42b for platelets (green), Clec4f for Kupffer cells (red), Glutamine Synthetase (GS) for central vein zone (purple). (C) Flow cytometric analysis of the percentage of P-selectin expression on platelets in SHR mice and mice at 0, 6, 24 h post-HS ( n = 6 mice per group). (D) GO (BP) analysis of cytokine-related pathways in platelets from SHR mice and at HS onset (HS0). (E, F) qPCR analysis of Il1b (E) and Casp1 (F) mRNA expression of platelets under heat stress in vitro ( n = 6 mice per group). (G) Flow cytometry analysis of caspase-1 levels in platelets from Tfr1 flox/flox mice, CKO- Tfr1 mice and CKO- Tfr1 mice infected with AAV9-DIO- Tfr1 , with or without ferroptosis inhibitor fer1 and DFO ( n = 6 mice per group). (H) ELISA analysis of IL-1 β levels in the supernatant of platelets under heat stress in vitro ( n = 6 mice per group). (I) ELISA analysis of IL-1 β levels in plasma from Tfr1 flox/flox mice, CKO- Tfr1 mice, and CKO- Tfr1 mice infected with AAV9-DIO- Tfr1 , with or without ferroptosis inhibitor fer1 and DFO ( n = 6 mice per group). (J) ELISA analysis of IL-1 β levels in PLP supernatant from sh- Zdhhc2, oe- Zdhhc2 , oe- Tfr1 , oe- Tfr1 -sh- Zdhhc2 MEG-01 cells under heat stress in vitro , with or without ferroptosis inhibitor fer1 ( n = 6 independent experiments). Mean ± SEM are shown, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: Coagulative factors, interleukin-1 β (IL-1 β ) and lipid peroxidation metabolites were measured by enzyme-linked immunosorbent assay in accordance with the manufacturer's instructions, respectively: IL-1 β (CSB-E08054m/E08053h, CUSABIO, Wuhan, China), IL-6 (CSB-E04639m, CUSABIO), IL-10 (CSB-E04594m-IS, CUSABIO), TNF- α (CSB-E04741m, CUSABIO) and 4-hydroxynonenal (4-HNE, CSB-E13412m/E16214h, CUSABIO).

Techniques: Derivative Assay, Immunofluorescence, Expressing, In Vitro, Flow Cytometry, Infection, Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Acta Pharmaceutica Sinica. B

Article Title: Palmitoylation of Tfr1 enhances platelet ferroptosis and liver injury in heat stroke

doi: 10.1016/j.apsb.2025.10.027

Figure Lengend Snippet: Tfr1 palmitoylation-mediated ferroptosis in platelets, accompanied by IL-1 β secretion, exacerbates liver injury in HS. The findings provide novel insights into thrombocytopenia and liver injury in HS, offering a potential therapeutic strategy to mitigate HS-induced thrombocytopenia, liver injury, and poor prognosis.

Article Snippet: Coagulative factors, interleukin-1 β (IL-1 β ) and lipid peroxidation metabolites were measured by enzyme-linked immunosorbent assay in accordance with the manufacturer's instructions, respectively: IL-1 β (CSB-E08054m/E08053h, CUSABIO, Wuhan, China), IL-6 (CSB-E04639m, CUSABIO), IL-10 (CSB-E04594m-IS, CUSABIO), TNF- α (CSB-E04741m, CUSABIO) and 4-hydroxynonenal (4-HNE, CSB-E13412m/E16214h, CUSABIO).

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Dual metabolic-inflammation modulation in MicroRNA@neutrophil-derived microvesicles achieve robust osteoarthritis therapy

doi: 10.1016/j.apsb.2025.09.020

Figure Lengend Snippet: miR140@MVs inhibit the pro-inflammatory effect of LPS-induced BMDM in vitro . (A) The intracellular location of Cy3-miR140@FITC-MVs in BMDM cells stimulated with LPS (100 ng/mL) for 24 h (M1 macrophages). miR140 is labeled with Cy3 (Red) and MVs are stained by FITC-Annexin V (Green). The nuclei were stained by Hoechst 33342 (Blue). Scale bar: 5 μm. (B) Representative flow cytometric analysis of BMDM treated with different formulations. The (C) M1 (CD86 + CD206 – ) and (D) M2 (CD86 − CD206 + ) phenotypes were analyzed by Flowjo. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, compared with M1 group. n = 3. No significance, ns; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗∗ P < 0.0001. (E) Western blotting of TLR4 in LPS-induced BMDM treated with different formulations. (F) The relative expression of the protein band was analyzed by Image J and normalized to its respective baseline controls. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. ∗ P < 0.05. The relative mRNA levels of (G) TNFα , (H) IL6 , (I) IL1β , (J) IL10 , (K) TGFβ in LPS-induced BMDM receiving different formulations. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, compared with the M1 group, n = 3. No significance, ns; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (L) The TNF α level in LPS-induced BMDM supernatant was detected by ELISA assay. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. No significance, ns; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. (F–I) Values are normalized to the M1 (baseline control) group.

Article Snippet: Cytokine concentration in the supernatant was quantified by mouse TNF α or mouse IL1 β enzyme-linked immunosorbent assay (ELISA) kits (Elabscience, USA).

Techniques: In Vitro, Labeling, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Acta Pharmaceutica Sinica. B

Article Title: Dual metabolic-inflammation modulation in MicroRNA@neutrophil-derived microvesicles achieve robust osteoarthritis therapy

doi: 10.1016/j.apsb.2025.09.020

Figure Lengend Snippet: Single miR140@MVs for prolonged OA treatment. (A) Schematic illustration of the administration regimen. (B) The relative miR140 level in the whole-knee joint of DMM mice treated with different formulations for 28 days. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. The normal group served as the control. No significance, ns. (C) Typical images of imageological examinations, histological analysis, TUNEL staining and immunohistochemical analysis. Red arrows: synovial inflammation; Yellow arrows: cartilage defect; Blue arrows: bone marrow edema; Red circles: periarticular osteophytes. M: meniscus; S: synovial; C: cartilage. Scale bars in H&E and safranine O-fast green staining: 100 μm; in TUNEL staining and immunohistochemical analysis: 50 μm. (D) Heatmap of variables of histological scoring in each group. (E) OARSI grades of the mice joints in each group. The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. The normal group served as the control. No significance, ns; ∗∗∗∗ P < 0.0001. (F–J) The relative mRNA levels of inflammatory factors in OA joints from mice after different treatments, including (F) TNFα , (G) IL6 , (H) IL1β , (I) IL10 and (J) TGFβ . The data are shown as mean ± SEM and analyzed by one-way ANOVA test with Tukey's correction, n = 3. The normal group served as control. No significance, ns; ∗∗ P < 0.01; ∗∗∗∗ P < 0.0001.

Article Snippet: Cytokine concentration in the supernatant was quantified by mouse TNF α or mouse IL1 β enzyme-linked immunosorbent assay (ELISA) kits (Elabscience, USA).

Techniques: Control, TUNEL Assay, Staining, Immunohistochemical staining