Canagliflozin inhibits inflammasome activation in diabetic endothelial cells – Revealing a novel calcium-dependent anti-inflammatory effect of canagliflozin on human diabetic endothelial cells

• Published in: Biomedicine & pharmacotherapy 2023-03, Vol.159, p.114228-114228, Article 114228
• Main Authors: Li, Xiaoling, Kerindongo, Raphaela P., Preckel, Benedikt, Kalina, Jan-Ole, Hollmann, Markus W., Zuurbier, Coert J., Weber, Nina C.
• Format: Article
• Language: English
• Subjects: Calcium; Canagliflozin (CANA); Canagliflozin - pharmacology; Caspase 1 - metabolism; Diabetes Mellitus; Endothelial Cells - metabolism; Extracellular signal regulated kinase (ERK) 1/2 phosphorylation; Humans; Inflammasomes - metabolism; Interleukin (IL)− 1β; Interleukin-1beta - metabolism; Intracellular calcium (Ca2+); NLR Family, Pyrin Domain-Containing 3 Protein - metabolism; Nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome; Reactive oxygen species (ROS); Reactive Oxygen Species - metabolism; Signal Transduction; Tumor Necrosis Factor-alpha
• ISSN: 0753-3322; 1950-6007
• DOI: 10.1016/j.biopha.2023.114228

Canagliflozin (CANA) shows anti-inflammatory and anti-oxidative effects on endothelial cells (ECs). In diabetes mellitus (DM), excessive reactive oxygen species (ROS) generation, increased intracellular calcium (Ca2+) and enhanced extracellular signal regulated kinase (ERK) 1/2 phosphorylation are crucial precursors for inflammasome activation. We hypothesized that: (1) CANA prevents the TNF-α triggered ROS generation in ECs from diabetic donors and in turn suppresses the inflammasome activation; and (2) the anti-inflammatory effect of CANA is mediated via intracellular Ca2+ and ERK1/2. Human coronary artery endothelial cells from donors with DM (D-HCAECs) were pre-incubated with either CANA or vehicle for 2 h before exposure to 50 ng/ml TNF-α for 2–48 h. NAC was applied to scavenge ROS, BAPTA-AM to chelate intracellular Ca2+, and PD 98059 to inhibit the activation of ERK1/2. Live cell imaging was performed at 6 h to measure ROS and intracellular Ca2+. At 48 h, ELISA and infra-red western blot were applied to detect IL-1β, NLRP3, pro-caspase-1 and ASC. 10 µM CANA significantly reduced TNF-α related ROS generation, IL-1β production and NLRP3 expression (P all  0.05). CANA and BAPTA both prevented intracellular Ca2+ increase in cells exposed to TNF-α (P both