Hy 10256

Migration assays were performed using 24-well transwell chamber with 8 μm pores (Costar, Bodenheim, Germany) to analyze the effect of CXCL10 on BMDM migration. Amounts of 2 × 105 BMDMs in 200 μl serum-free DMEM high glucose medium were seeded in the upper wells, and 600 μl serum-free DMEM medium with or without CXCL10 (10, 50, 100 ng/ml) was added to the lower wells. In inhibiting experiments, BMDMs were pretreated with AMG487 (1 μM), Erk1/2 inhibitor (PD98059, HY-12028, MedChem Express), or p38 MAPK inhibitor (SB203580, HY-10256, MedChem Express) for 1 h. After 24 h of incubation, cells attached on the lower surface of the membrane were fixed with 100% methanol for 15 min and stained with 0.05% crystal violet staining.

A total of 48 SD rats were randomly divided into the sham group (n = 12) and the PH-LHD group (n = 36). Corresponding to different inhibitors and mechanical strength used to treat pulmonary venous rings, the sham group was further divided into two subgroups: Sham 1 (S1, mechanical strength: 0 g) and Sham 2 (S2, mechanical strength: 2.0 g); and the PH-LHD group was also further divided into 2 subgroups: Model 1 (M1, banding+0 g), Model 2 (M2, banding+2.0 g). In addition, a broad spectrum inhibitor of MEK1 and MEK2 (U0126, HY-12031 MedChem Express, USA), p38 MAPK inhibitor (SB203580, HY-10256 MedChem Express, USA), SAC inhibitor (Streptomycin, HY-B0472 MedChem Express, USA) and broad-spectrum inhibitor of JNK1, JNK2, and JNK3 (SP600125, HY-12041 MedChem Express, USA) were dissolved in dimethyl sulfoxide (DMSO, HY-10999 MedChem Express, USA), and diluted with K-H solution to 250 μmol/L, 200 μmol/L, 1000 μmol/L, and 250 μmol/L, respectively [12 (link)]. Specifically, the pulmonary veins were immersed in the K-H solution containing the corresponding inhibitors (SB203580, SP600125, U0126 or Streptomycin) at 37°C under the condition of continuous oxygen infusion for 60 mins, and then was stretched with 2.0 g mechanical strength for 60 mins.