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: To obtain M1 phenotype macrophages, RAW264.7 was treated with LPS (100 ng/mL) (Invitrogen, CA, USA) for 24 h. L929 mouse fibroblast cells were purchased from the ATCC and grown at 37 °C with 5% CO 2 for 7 days in DMEM supplemented with 10% FBS.
Techniques: In Vitro, Labeling, Staining, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Control
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