Fig. 7: BDP-12 demonstrates diminished mitogenicity and strong anti-inflammatory effect via antagonizing TLR4.

A Mice treated with rBDNF or BDP-12 for 3 d underwent immunohistochemistry staining for PCNA in lung tissues. Quantification of PCNA⁺ cells was shown in the right. B MLE-12 cells were treated with 10 nM rBDNF or 2 μM BDP-12 for 0.5 h. Then the phosphorylated TrkB, total TrkB, and GAPDH were assessed by immunoblotting. C MLE-12 cells were treated with gradient rBDNF (0–20 nM) or BDP-12 (0–5 μM) for 24 h. Relative cell amounts were quantified using the CCK8 assay (n = 4). D Primary macrophages were pre-treated with 2, 4 μM BDP-12 and then exposed to LPS for 0.5 h. Then the TLR4 was immunoprecipitated and the associated MD2 level was detected using immunoblotting. E–G WT and Tlr4^-/- macrophages were pre-treated with BDP-12 (2, 4 μM) and then exposed to LPS for 6 h. Relative mRNA levels of Tnf (E), Il6 (F), and Il1b (G) were measured. H Schematic illustration of mice undergoing ALI modeling and BDP-12 injection. I Representative H&E-stained images of lung tissues. J Quantification of lung injury scores for (I). K Total protein levels in BALF samples. L Lung wet/dry weight ratio. M Total cell counts in BALF samples. N Survival curve of mice with or without BDP-12 injection in an E.coli-induced sepsis model. O The level of MD2-TLR4 complex formation in lung tissues was assessed by Co-IP. P Heatmap showing mRNA levels of pro-inflammatory factors Tnf, Il6, Il1b, Ccl2, Ifna1, Ifnb1, and Ifng in lung tissues. Q, R Serum (Q) and BALF (R) levels of TNF-α, IL-6, and IFN-α in the ALI mouse model. Data are represented as mean ± SEM by one-way ANOVA with Tukey’s test. n = 4 (C, E–G), or n = 6 (A, J–M, P–R) per group. Source data are provided as a Source Data file.