Journal: Acta Pharmaceutica Sinica. B

Article Title: Inhalable songorine-integrated lipid nanomedicine for targeted ARDS therapy via repairing endothelial barrier and inactivating NLRP3 inflammasome

doi: 10.1016/j.apsb.2025.10.048

Figure Lengend Snippet: Effects of Son-lipo in repairing the endothelial barrier, inhibiting inflammation, and mitigating cell apoptosis in vitro . (A) Immunofluorescent staining of tight junction factor ZO-1 in HUVEC cells treated with different formulations. Red, ZO-1. Blue, Hoechst33342. Scale bar = 20 μm. Protein expression of ZO-1 by (B) Western blot and (C) statistical analysis in HUVEC cells ( n = 3). (D) The mRNA expression of tight junction factors, TJP1 , CDH5 , and CLDN5 , in HUVEC cells by RT-qPCR ( n = 3). (E) The mRNA expression of inflammatory factors, Il1b , Il6, and Tnfa , in RAW264.7 cells by RT-qPCR ( n = 4). Internal control, α -Tubulin. The apoptosis rate of endothelial cells was detected by (F) flow cytometry and (G) analyzed by FlowJo in HUVEC cells ( n = 3). Data are represented as mean ± SD. ∗ P < 0.05 vs Control. # P < 0.05 vs Model. † P < 0.05.

Article Snippet: The cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.5% Triton X-100 for 10 min, and finally blocked with 5% BSA for 1 h. Subsequently, the cells were incubated with tight junction factor ZO-1 antibody (1:1000, Proteintech, 21773-1-AP, Wuhan, China) overnight.

Techniques: In Vitro, Staining, Expressing, Western Blot, Quantitative RT-PCR, Control, Flow Cytometry

Journal: Acta Pharmaceutica Sinica. B

Article Title: Inhalable songorine-integrated lipid nanomedicine for targeted ARDS therapy via repairing endothelial barrier and inactivating NLRP3 inflammasome

doi: 10.1016/j.apsb.2025.10.048

Figure Lengend Snippet: Son-lipo ameliorated the phenotypes of LPS-induced ARDS in vivo . (A) Schedule of the construction and treatments of ARDS mouse model. (B‒E) The results of pulmonary function indexes (EF50, Penh, TV/body weight, and EEP) by non-invasive whole-body plethysmography ( n = 4‒5). (F) Immunohistochemistry staining of ZO-1 and CLAUDIN-5 protein (Scale bar = 100 μm), and immunofluorescent staining of VE-CADHERIN protein (red, VE-CADHERIN; blue, Hoechst33342; Scale bar = 20 μm). (G) Statistical analysis of ZO-1 and CLAUDIN-5 protein expression in F ( n = 3). (H) The mRNA expressions of tight junction factors, Tjp1 , Cldn5 , and, Cdh5 in lung tissues by RT-qPCR analysis ( n = 3). Internal control, α -Tubulin. (I) HE staining of lung tissues. Black arrows, the degree of inflammatory cell infiltration and destroyed alveoli structures. Red arrows, alveoli wall. Blue arrows, the degree of alveoli leakage. Scale bar = 1000 μm for the upper panels and 100 μm for the lower panels. (J) Pathological scores of HE staining results ( n = 5‒6). (K) Total cell number and (L) protein content in BALF ( n = 4‒5). (M) Lung index of mice ( n = 5). (N‒P) The change in folds of mRNA levels of inflammatory factors, Il1b , Tnfa , and Nos2 , in lung tissues ( n = 3). (Q, R) The ELISA analysis of IL-6 and TNF- α levels ( n = 4). (S) TUNEL staining and (T) statistics of positive cells in lung tissues ( n = 3). Blue, Hoechst33342. Scale bar = 50 μm. Data are represented as mean ± SD. ∗ P < 0.05 vs Control. # P < 0.05 vs Model. † P < 0.05.

Article Snippet: The cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.5% Triton X-100 for 10 min, and finally blocked with 5% BSA for 1 h. Subsequently, the cells were incubated with tight junction factor ZO-1 antibody (1:1000, Proteintech, 21773-1-AP, Wuhan, China) overnight.

Techniques: In Vivo, Immunohistochemistry, Staining, Expressing, Quantitative RT-PCR, Control, Enzyme-linked Immunosorbent Assay, TUNEL Assay