Fig 2: ECM1-HF-MSCs inhibit the TGF-ß/Smad signaling pathway and activation of HSCs in vitro.A The quiescent murine hepatic stellate cell line JS1. B Activated JS1 cells. Scale bar = 50 µm. C Western blotting analysis of a-SMA, TGF-ß, and p-Smad2/3 in all groups. D–F Semiquantitative analysis of the pathway proteins and a-SMA. Data are shown as the means ± SDs (*p < 0.05, **p < 0.01, ***p < 0.001).

Fig 3: ECM1-HF-MSCs show a better effect in LC treatment through inhibiting the activation of HSCs and the TGF-ß/Smad signaling pathway in vivo.A, B HE and Masson staining in each group. Scale bar = 100 µm. C Area of collagen fibers in each group. D Ishak scores show the degree of LC in all groups. E–G Changes in the serological indices of ALT, AST and ALP. H–K Western blotting and semiquantitative analysis of a-SMA, p-Smad2/3, and TGF-ß in all groups. Data are shown as the means ± SDs (*p < 0.05, **p < 0.01, ***p < 0.001).

Fig 4: A maternal high-fat diet (HFD) is associated with thickening of the bronchial and vascular smooth muscle cell (SMC) layer as well as bronchial obstruction and pulmonary arterial hypertension beyond infancy.A Representative images of the lungs of offspring at postnatal day 70 (P70). Immunohistochemistry for a-smooth muscle actin (aSMA) was performed to assess the bronchial SMC layer in the offspring of dams fed a HFD or standard diet (SD). B Quantitative measurement of the bronchial SMC layer in bronchi with diameters <150 µm and 150–200 µm. C Assessment of respiratory airway resistance using body plethysmography at P70. D–F Representative images of the lungs of offspring at P70. Dual staining for aSMA and von Willebrand factor (vWF) (D) was used to assess the muscularization of microvessels (20–100 µm) (E) and the medial wall thickness of microvessels (diameters: 20–70 µm and 70–150 µm) (F). G–K Echocardiographic assessment of heart rate (G), cardiac output (CO, H), stroke volume (SV, I), right ventricular internal diameter (RVID, J) and tricuspid annular plane systolic excursion (TAPSE, K). L, M Hemodynamic measurement of right ventricular systolic pressure (RVSP, L), and total pulmonary vascular resistance index (TPVRI, M). N Echocardiographic assessment of the left ventricular (LV) ejection fraction (EF). O, P Measurement of the systolic blood pressure (SBP) and diastolic blood pressure using invasive catheterization. Q qRT-PCR assessment of Nppa expression in the heart at P70. R Immunoblot analysis of aSMA protein in total lung homogenates at P21; ß-actin was used as a loading control. A summary of the aSMA abundance relative to ß-actin is displayed next to the immunoblot. S qRT-PCR assessment of Acta2 expression in laser-microdissected bronchi and vessels at P21; 18 s rRNA served as a housekeeping gene. Data are shown as mean ± standard error of the mean. B, C, E–S SD n = 4–17, HFD n = 6–9. Statistical analyses were performed using the two-sided Mann–Whitney test or two-sided Student’s t test; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Gray = standard diet; white = high-fat diet. A detailed list of sample sizes and P value per graph is provided in the Supplemental Material. Source data are provided as a Source Data file.