Figure 6.

Therapeutic STIMATE⁺ ADE in the BLM-induced injurious pulmonary fibrosis model. (A) The protocol of rmSP-A/mtDNA-coated agarose beads stimulate type II alveolar epithelial MLE-12 cells to secrete sufficient STIMATE⁺ ADE for translational treatment of BLM-induced pulmonary fibrosis. (B) Confocal images of apoptosis-labeled TUNEL and mouse AEC-IIs-labeled SFTPC of BLM-induced mouse lung sections after 14 days of induction with 0.125 U/kg BLM. (C) ELISA detected the level of STIMATE⁺-ADE in the BALF of mice after 14 days of induction with 0.125 U/kg BLM (n = 8). (D) Mouse lung immunofluorescence sections fluorescent merge of Siglec F-labeled AMs and STIMATE-GFP ADE. Administration of inhaled atomized particles to mice (twice a week, 10⁹ to 10¹⁰ particles per mouse) and mouse AEC-IIs tracheal colonization (2×10⁶ cells per mouse) after intraperitoneal administration of 0.125 U/kg bleomycin. (E) Flow cytometry to detect the ratio of M1-like and M2-like TRAMs in mouse BALF after intraperitoneal administration of 0.125 U/kg bleomycin, preadministration of inhaled atomized particles to mice or mouse AEC-IIs tracheal colonization (n = 6). (F and G) Seahorse monitoring of mitochondrial respiration and glycolysis capacity in BALF TRAMs after 7 days and 14 days of BLM intraperitoneal administration in the presence or absence of inhaled atomized particles to mice or mouse AEC-IIs tracheal colonization (n = 6). (H and I) H&E staining and a pathological score of paraffin-embedded lung sections after 7 days and 14 days of BLM intraperitoneal injection in the presence or absence of inhaled atomized particles to mice or mouse AEC-IIs tracheal colonization. 400× microscope (n = 6), whole lung. (J and K) Hydroxyproline content (right lung) and respiratory system compliance of the lung after 7 days and 14 days of BLM intraperitoneal injection in 4-week-old mice in the presence or absence of inhaled atomized particles to mice or mouse AEC-IIs tracheal colonization (n = 6). (L) BLM induces long-term survival in mice with chronic pulmonary fibrosis in the presence or absence of inhaled atomized particles or mouse AEC-IIs tracheal colonization (n = 14). Data are shown as the mean ± SD, *p < 0.05 and **p < 0.01 by paired Student's t test and Gehan-Breslow-Wilcoxon test (FIGURE W) compared to WT mice or untreated mice.