Tectorigenin

Kinase experiment:

HUVECs grown to confluence in 24-well plates are pretreated with Tectorigenin (0.1, 1, 10 μM), salicylate (5 mM) or GSH (1 mM) for 30 min, then stimulated with Palmitic acid (PA) (100 μM) for further 12 h in serum-free medium, and the medium is then collected on ice. The levels of TNF-α and IL-6 in the supernatant are assayed with commercial ELISA Kits[1].

Cell experiment:

Cell viability is assessed by MTT method. Briefly, cells are seeded in 96-well plate at a density of 1×104 cells/well. After 24 h incubation, Tectorigenin at different concentrations is added to the cells while only DMSO (solvent) is added as a negative control. After growing for 12, 24 and 48 h, cells are incubated with MTT (0.5 mg/mL) for 4 h at 37°C. During this incubation period, water-insoluble formazan crystals are formed, which are dissolved by the addition of 100 μL/well DMSO. The optical densities at 570 nm are measured using an enzyme-linked immunosorbent assay plate reader. Wells containing culture medium and MTT but no cells act as blanks[2].

References:

[1]. Wang Q, et al. Tectorigenin Attenuates Palmitate-Induced Endothelial Insulin Resistance via Targeting ROS-Associated Inflammation and IRS-1 Pathway. PLoS One. 2013 Jun 19;8(6):e66417.
[2]. Jiang CP, et al. Pro-apoptotic effects of tectorigenin on human hepatocellular carcinoma HepG2 cells. World J Gastroenterol. 2012 Apr 21;18(15):1753-64.