Corosolic acid ameliorates vascular remodeling in pulmonary arterial hypertension via the downregulation of STAT3 signaling

Up-regulated expression of two-pore domain K+ channels, KCNK1 and KCNK2, is involved in the proliferation and migration of pulmonary arterial smooth muscle cells in pulmonary arterial hypertension

“…Cell viability increases in a time-dependent manner in both normal-PASMCs (A 450 = 0.393 ± 0.028 at 0 h, 0.775 ± 0.020 at 24 h, 0.906 ± 0.052 at 48 h, and 1.004 ± 0.067 at 72 h, n = 4) and IPAH-PASMCs (0.441 ± 0.008 at 0 h, 0.909 ± 0.019 at 24 h, 1.184 ± 0.042 at 48 h, and 1.330 ± 0.067 at 72 h, n = 5) (Figure 4A). The growth of IPAH-PASMCs at 48 and 72 h was markedly greater than that of normal-PASMCs ( p = 0.020 and p = 0.004, respectively), which is consistent with previous findings (23,26,27).…”

“…Animal experiments were approved by the Ethics Committees of Nagoya City University (H30-P-1) and Aichi Medical University (2019–15). To generate MCT-PH rats ( 20 – 22 ), rats (Sprague-Dawley, male, 4 weeks old, 100–110 g; Japan SLC, Hamamatsu, Japan) were subcutaneously injected with vehicle (saline) or MCT (60 mg/kg; MilliporeSigma) and bred for 3 weeks ( 23 ). To obtain SuHx-PH rats ( 20 – 22 ), rats (Sprague-Dawley, male, 6 weeks old, 220–250 g; Japan SLC) were subcutaneously injected with vehicle (saline) or Sugen 5416 (20 mg/kg; MedChemExpress, Monmouth Junction, NJ, USA) and bred in a hypoxic chamber (10% O 2 ) with a ProOx110 O 2 controller (Biospherix, Parish, NY, USA) for 3 weeks and thereafter in a normoxic chamber for 2 weeks.…”

“…Protein fraction was extracted using RIPA buffer (for human PASMCs) and T-PER Tissue Protein Extraction Reagent (for rat/mouse pulmonary arterial smooth muscles (PASMs); Thermo Fisher Scientific) ( 23 ). Extracted protein (20 μg/lane) was applied to an acrylamide gel (8%) and transferred to an Immobilon-P PVDF membrane (MilliporeSigma).…”

“…Human PASMCs (3 × 10 3 cells/well) were seeded on a 96-well plate (Falcon #353075, Corning, Corning, NY, USA) and cultured at 37°C for 6 h ( 23 ). Thereafter, they were treated with culture medium containing FBS (10%) and the vehicle [dimethyl sulfoxide (DMSO)] or drug for 48 h. Cell viability was assessed using Cell Counting Kit-8 (Dojindo Laboratories) based on the WST-8 assay.…”

“…Human PASMCs (5 × 10 4 cells/well) were seeded on a 24-well Transwell insert with a membrane pore size of 8 μm (#3422, Corning) ( 23 ). Thereafter, they were treated with culture medium containing FBS (1%) in the upper chamber and that containing FBS (10%) and the vehicle (DMSO) or drug in the lower chamber for 24 h. Transwell inserts were fixed in paraformaldehyde (4%) and stained with crystal violet (1%; Fujifilm Wako Pure Chemical).…”