Fluorescent probe 2 7 dichlorofluorescein diacetate dcfh da

THC was purchased from Shanghai Macklin Biochemical Co. Ltd. (Shanghai, China) with a purity of 98% tested by high-performance liquid chromatography (HPLC). STZ was purchased from Sigma-Aldrich (St. Louis, MO, USA). The fluorescent probe 2′,7′-dichlorofluorescein diacetate (DCFH-DA) used to detect intracellular ROS generation was purchased from the Beyotime Institute of Biotechnology (Shanghai, China). Dihydroethidium (DHE; for detecting ROS generation in cardiac tissues) were purchased from Invitrogen (Carlsbad, CA, USA). A primary antibody against SOD2 (sc-30080) was obtained from Santa Cruz Biotechnology (CA, USA). Primary antibodies against T-Smad3 (SC101154), p-Smad3 (8760), and β-actin (4970) were all purchased from Cell Signaling Technology (Boston, MA, USA). Primary antibodies against SIRT1 (ab110304), acetylated SOD2 (Ac-SOD2, ab137037), α-SMA (ab5694), TGFβ1 (ab92486), collagen І (ab90395), and collagen III (ab7778) were purchased from Abcam (Cambridge, MA, USA). The rabbit anti-goat and goat anti-mouse secondary antibodies were obtained from the Zhongshan Company (Beijing, China).

BAK (purity ≥ 98%) was purchased from Winherb Medical Co. (Shanghai, China). Streptozotocin (STZ) and EX527 were purchased from Sigma-Aldrich (St. Louis, MO, USA). The In Situ Cell Death Detection Kit was purchased from Roche Biochemicals (Mannheim, Germany) to detect the cell apoptosis. Kits for determining glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) activities, and malondialdehyde (MDA) content were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The fluorescent probe 2′,7′-dichlorofluorescein diacetate (DCFH-DA) was purchased from Beyotime Institute of Biotechnology (Shanghai, China) to detect intracellular ROS generation. Dihydroethidium (DHE) was purchased from Invitrogen (Carlsbad, CA, USA) for detecting ROS generation in the myocardium. Primary antibodies against p-Smad3, t-Smad3, and β-actin were purchased from Cell Signaling Technology (Boston, MA, USA). Primary antibodies against SIRT1, Nrf2, transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA), and Histone H3 were purchased from Abcam (Cambridge, MA, USA). The rabbit anti-goat and goat anti-mouse secondary antibodies were obtained from Zhongshan Company (Beijing, China).