Fig. 3.
Intratracheal-delivered rISM1 alleviates emphysema in Ism1−/− and CS-exposed mice. (A) Quantification of AMs from Ism1−/− lung under PBS, rISM1, and liposome-clodronate (CLO) treatments. (B) IF staining and quantifications for GRP78highCD68+ AMs, nuclei are stained by DAPI. (C) Representative H&E-stained lungs of 2-mo-old FVB/NTac Ism1−/− mice after vehicle (PBS), 1 μg rISM1, 5 μg rISM1, and CLO treatments. (D) Quantifications of MLI and (E) FEV100/FVC of untreated and treated mice groups in A. (F) Experimental design of 2-wk and (G) 8-wk CS-induced COPD model in WT BALB/cAnNTac (WT BALB/c) mice. Room air–exposed WT BALB/c mice (Sham) and CS-exposed WT BALB/c mice (CS) with vehicle (PBS) or rISM1 (10 µg rISM1) treatments at frequency and intervals indicated. (H) Quantifications of BALF cells from experimental groups in F. (I) H&E stained lungs of experimental groups in (G) depicting immune cell infiltration. n = 5 mice per group. (J) Confocal fluorescent microscopy demonstrating cell-surface expression of GRP78 in AMs in Sham and CS mouse lungs in G. (K) IF staining and quantifications of GRP78high AMs and (L) total AMs for experiment groups in G. (M) Active MMP-12 detected by Western blot and their quantifications for experiment groups in G. (N) MLI and (O) FEV100/FVC of experimental groups in G. Data are mean ± SD and were analyzed by two-group, two-tailed Student’s t test (B, D, and E: WT and Ism1−/− comparisons; and K) and one-way ANOVA with Tukey’s post hoc test (A, D, E, H, and L–O). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001, #: no significant difference compared to Sham group. n = 3 to 5 mice per group. (Scale bars: 20 μm for B and K, 50 μm for C and I.) Data from A–E are representatives of twice-repeated experiments with similar results. Data from H is representative of one independent experiment. Data from I–O are independent experiments using the same experimental groups of mice in L.