Alagebrium chloride (ALT711)

Cell experiment:

Human aortic endothelial cells (HAECs) are seeded on decellularized matrices derived from the abdominal aorta (AAo) of Zucker lean (ZL), obese (ZO), and diabetic (ZD) rats with or without Alagebrium (ALT-711) (20 μg/mL in Dulbecco's PBS with 1× antibiotic-antimycotic). Experiments are performed when cells reach 80 to 90% confluence. Flow chambers are sealed to the HAEC monolayers via a vacuum network. Flow is driven by a Masterflex L/S peristaltic pump in a humidified chamber heated to 37°C for 4 h. Leibovitz-15 medium, supplemented with 10% FBS, endothelial BulletKit, and 1× antibiotic-antimycotic solution, is used as the flow medium to maintain pH in the absence of CO2[2].

Animal experiment:

RAGE apoE mice are randomized to be treated with Alagebrium (1 mg/kg/day by gavage), or no treatment (n=20/group). After 20 weeks of diabetes, mice are placed into individual metabolic cages for 24 h and urine is collected. Body weight as well as fluid and food intake are recorded. Urinary albumin excretion is estimated in urine samples by a mouse albumin enzyme-linked immunosorbent assay (ELISA) kit according to the kit protocol. Urinary and serum creatinine concentrations are measured by high-performance liquid chromatography (HPLC). Systolic blood pressure is assessed by a noninvasive tail cuff method in conscious mice at the end of the study[1].

References:

[1]. Watson AM, et al. Alagebrium reduces glomerular fibrogenesis and inflammation beyond preventing RAGEactivation in diabetic apolipoprotein E knockout mice. Diabetes. 2012 Aug;61(8):2105-13.
[2]. Wang H, et al. Alagebrium inhibits neointimal hyperplasia and restores distributions of wall shear stress by reducing downstream vascular resistance in obese and diabetic rats. Am J Physiol Heart Circ Physiol. 2015 Oct;309(7):H1130-40.