Journal: Materials Today Bio

Article Title: Naringin nanoparticles alleviate RA-ILD pulmonary fibrosis by targeting 14-3-3ζ to inhibit LYVE1 + macrophages TGF-β1 secretion

doi: 10.1016/j.mtbio.2026.102982

Figure Lengend Snippet: NAR promotes LYVE1 expression in lung macrophages. (A) Differentially expressed genes of LYVE1 were screened out from lung tissue of CIA-ILD mice and lung tissue of NAR + CIA-ILD mice. (B) The expression of LYVE1 protein levels in the human tissue was analyzed by western blotting (n = 4). (C) The expression of LYVE1 protein levels in the mice lung tissue was analyzed by western blotting (n = 3). (D) Representative images showing immunofluorescence staining for LYVE1 (RED) in mice lung tissue, scale bar: 100 μm, 50 μm. (E) BLM in alveolar lavage fluid of RA-ILD patients were sifted by flow cytometry. (F-G) The expression of LYVE1 mRNA and protein levels in BLM induced by NAR at different concentrations for 48 h were analyzed by RT-qPCR and western blotting (n = 3). (H) Representative confocal microscopy images showing immunofluorescence staining for LYVE1 (RED) and CD68 (GREEN) in BLM induce by 150 μM NAR for 48 h, scale bar: 10 μm ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To conduct the staining procedure, employ the following antibodies: CD68 (santa cruz biotechnology, inc) and LYVE1 (fitzgerald industries international, LYVE1 antibody).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Flow Cytometry, Quantitative RT-PCR, Confocal Microscopy

Journal: Materials Today Bio

Article Title: Naringin nanoparticles alleviate RA-ILD pulmonary fibrosis by targeting 14-3-3ζ to inhibit LYVE1 + macrophages TGF-β1 secretion

doi: 10.1016/j.mtbio.2026.102982

Figure Lengend Snippet: NAR targets 14-3-3ζ to promote LYVE1 expression in lung macrophages. (A) The target prediction process of NAR acting on BLM. (B) The expression of 14-3-3ζ protein levels in the human lung tissue was analyzed by western blotting (n = 4). (C) The expression of 14-3-3ζ protein levels in lung tissue of experimental mice were analyzed by western blotting (n = 3). (D) Representative images showing immunofluorescence staining for 14-3-3ζ (RED) in mice lung tissue, scale bar: 100 μm, 50 μm. (E) The predictive model of NAR binding to 14-3-3ζ. (F) Representative confocal microscopy images showing immunofluorescence staining for LYVE1 (RED) and 14-3-3ζ (GREEN) in BLM induce by 150 μM NAR for 48h, scale bar: 10 μm. (G) NAR binding affinity to 14-3-3ζ was determined by MST. (H-I) Binding ability of NAR to 14-3-3ζ at different temperatures (H) and different doses (I) was analyzed by CETSA. (J) BLM were transfected by OE-14-3-3ζ mRNA, and were induced with 150 μM NAR for 48 h. The expression of LYVE1/14-3-3ζ protein levels were analyzed by western blotting. (n = 3); (K) Luciferase activity assay was conducted on BLM transfected by OE-14-3-3ζ mRNA and treated with 150 μM NAR for 48 h (n = 3). (L-M) BLM were transfected by si-14-3-3ζ RNA, and were induced with 150 μM NAR for 48 h. The expression of LYVE1/14-3-3ζ mRNA and protein levels were analyzed by RT-qPCR and western blotting (n = 3). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To conduct the staining procedure, employ the following antibodies: CD68 (santa cruz biotechnology, inc) and LYVE1 (fitzgerald industries international, LYVE1 antibody).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Binding Assay, Confocal Microscopy, Transfection, Luciferase, Activity Assay, Quantitative RT-PCR

Journal: Materials Today Bio

Article Title: Naringin nanoparticles alleviate RA-ILD pulmonary fibrosis by targeting 14-3-3ζ to inhibit LYVE1 + macrophages TGF-β1 secretion

doi: 10.1016/j.mtbio.2026.102982

Figure Lengend Snippet: NAR-induced LYVE1 + macrophages suppress EMT in pulmonary epithelial cells and FMT in lung fibroblasts. (A) CD68 + LYVE1 - BLM and CD68 + LYVE1 + BLM were sorted by flow cytometry. (B-C) After LYVE1 + BLM and LYVE1 - BLM were induced by 150 μM NAR for 48 h, analyzed the expression of LYVE1 and 14-3-3ζ mRNA and protein levels by RT-qPCR and western blotting (n = 3). (D) The flowchart of co-culture of NAR-induced BLM and A549/HFL1 cells. (E-F) Scratch experiment, added LYVE1 - BLM and LYVE1 + BLM which were induced by NAR, and set up Ctrl. Scale bar: 750 μm (n = 3). (G-H) Transwell invasion experiment, LYVE1 - BLM and LYVE1 + BLM incubated with 150 μM NAR for 48h were added to the lower chamber of 24-well plate and Ctrl was set, and A549 cells were added to the upper chamber. scale bar: 750 μm (n = 3). (I-J) LYVE1 - BLM/LYVE1 + BLM and A549 cells were co-cultured with 150 μM NAR for 48 h, and analyzed the expression of EMT-related (E-cadherin, vimentin) mRNA and protein levels of A549 cells by RT-qPCR and western blotting (n = 3). (K-L) LYVE1 - BLM/LYVE1 + BLM and HFL1 cells were co-cultured with 150 μM NAR induction for 48h, and analyzed the expression of FMT-related (fibronectin, COL1A1, α-SMA) mRNA and protein levels of HFL1 cells by RT-qPCR and western blotting (n = 3). (M) Representative images showing immunofluorescence staining for E-cadherin, vimentin, fibronectin, COL1A1 and α-SMA (RED) in mice lung tissue, scale bar: 100 μm, 50 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To conduct the staining procedure, employ the following antibodies: CD68 (santa cruz biotechnology, inc) and LYVE1 (fitzgerald industries international, LYVE1 antibody).

Techniques: Flow Cytometry, Expressing, Quantitative RT-PCR, Western Blot, Co-Culture Assay, Incubation, Cell Culture, Immunofluorescence, Staining

Journal: Materials Today Bio

Article Title: Naringin nanoparticles alleviate RA-ILD pulmonary fibrosis by targeting 14-3-3ζ to inhibit LYVE1 + macrophages TGF-β1 secretion

doi: 10.1016/j.mtbio.2026.102982

Figure Lengend Snippet: NAR reduces TGF-β1 secretion by enhancing LYVE1 expression in macrophages. (A) The expression of macrophage-related pulmonary fibrosis cytokines TGF-β1, TGF-β2, CXCL2, and CXCL4-related mRNA in LYVE1 - BLM and LYVE1 + BLM were analyzed by RT-qPCR (n = 3). (B-C) The expression of TGF-β1 protein levels in human lung tissue (n = 4) and the expression of TGF-β1 protein levels in the mice lung tissue was analyzed by western blotting (n = 3). (D) Representative images showing immunofluorescence staining for TGF-β1 (RED) in mice lung tissue, scale bar: 100 μm, 50 μm. (E) The expression of TGF-β1 in serum of Ctrl, RA, and RA-ILD patients by ELISA (n = 20). (F) The expression of TGF-β1 in BLM supernatant by ELISA (n = 3). (G-H) The expression of TGF-β1 mRNA and protein levels BLM treated with various concentrations of NAR for 48 h by RT-qPCR and western blotting (n = 3), (I) Representative confocal microscopy images showing immunofluorescence staining for LYVE1 (RED) and TGF-β1 (GREEN) in BLM induce by 150 μM NAR for 48 h, scale bar: 10 μm. (J) The expression of LYVE1, 14-3-3ζ and TGF-β1 protein levels in LYVE1 - BLM and si-LYVE1 RNA interfered LYVE1 + BLM induced by 150 μM NAR for 48h were analyzed by western blotting. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To conduct the staining procedure, employ the following antibodies: CD68 (santa cruz biotechnology, inc) and LYVE1 (fitzgerald industries international, LYVE1 antibody).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Confocal Microscopy

Journal: Materials Today Bio

Article Title: Naringin nanoparticles alleviate RA-ILD pulmonary fibrosis by targeting 14-3-3ζ to inhibit LYVE1 + macrophages TGF-β1 secretion

doi: 10.1016/j.mtbio.2026.102982

Figure Lengend Snippet: NAR alleviates pulmonary fibrosis by reducing TGF-β1 secretion from LYVE1 + macrophages. (A-B) Scratch experiment, added LYVE1 + BLM which incubated with 150 μM NAR for 48 h in the upper chamber, TGF-β1 5 ng/ml was added to the lower chamber. scale bar: 750 μm (n = 3). (C-D) Transwell invasion experiment, LYVE1 + BLM which incubated with 150 μM NAR for 48 h was added to the lower chamber of a 24-well plate. A549 cells were added to the upper chamber, with the addition of TGF-β1 5 ng/ml scale bar: 750 μm (n = 3). (E-H) LYVE1 + BLM which were induced with 150 μM NAR for 48h were co-cultured with A549/HFL1 cells for 48 h, with the addition of TGF-β1 5 ng/ml. The expression of EMT/FMT-related mRNA and proteins levels in A549/HFL1 cells were analyzed by RT-qPCR and western blotting (n = 3). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: To conduct the staining procedure, employ the following antibodies: CD68 (santa cruz biotechnology, inc) and LYVE1 (fitzgerald industries international, LYVE1 antibody).

Techniques: Incubation, Cell Culture, Expressing, Quantitative RT-PCR, Western Blot

Journal: Materials Today Bio

Article Title: Naringin nanoparticles alleviate RA-ILD pulmonary fibrosis by targeting 14-3-3ζ to inhibit LYVE1 + macrophages TGF-β1 secretion

doi: 10.1016/j.mtbio.2026.102982

Figure Lengend Snippet: NAR promotes the expression of LYVE1 to inhibit TGF-β1 secretion by targeting 14-3-3ζ in macrophages, which can alleviate pulmonary fibrosis in patients with RA-ILD.

Article Snippet: To conduct the staining procedure, employ the following antibodies: CD68 (santa cruz biotechnology, inc) and LYVE1 (fitzgerald industries international, LYVE1 antibody).

Techniques: Expressing