Mmp 9 inhibitor 1

Details of reagents administration are presented (Table 1). MMP-9 inhibitor I (sc-311437, Santa Cruz Biotechnology) was dissolved in DMSO to obtain a 5 mM stock solution, and applied to cells 12 h before addition of H₂O₂ (final concentration, 5 µM). SB431542 [TGF-β RI inhibitor, (HY-10431, MedChemExpress, New Jersey, USA)] and LY2109761 [TGF-β RI/II inhibitor, (HY-12075, MedChemExpress)] were dissolved in DMSO to prepare a 20 mM and 10 mM stock solution, respectively. SB431542 and LY2109761 were applied to cells (final concentration, 10 µM and 5 µM, respectively) which had been exposed to 400 µM H₂O₂ for 24 h. TGF-β1 RP (APA124Hu01, USCN Business Co., Ltd., Wuhan, China) was dissolved in PBS to prepare a 10 µg/ml stock solution and applied to cells at 15 ng/ml for 96 h. SB203580 (S8307-1MG, Sigma Aldrich; Merck KGaA, Darmstadt, Germany), a cytokine-suppressive anti-inflammatory drug, was used as a p38 MAPK inhibitor in this study. It was also dissolved in DMSO at 30 mM to prepare a stock solution. The cells were pretreated with SB203580 at 30 μM for 4 h before the addition of H₂O₂. R)-(+)-Limonene with a purity of 97% was purchased from Sigma Aldrich and diluted using culture medium prior to use. Cells were pretreated with R)-(+)-Limonene at 1000 µM for 4 h prior to addition of H₂O₂.