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

  • Biomed Pharmacother. 2023 Jan 7;159:114228. doi: 10.1016/j.biopha.2023.114228.
Xiaoling Li  1 ,  Raphaela P Kerindongo  2 ,  Benedikt Preckel  3 ,  Jan-Ole Kalina  4 ,  Markus W Hollmann  5 ,  Coert J Zuurbier  6 ,  Nina C Weber  7
Affiliations
  • 1. Amsterdam, University Medical Centers, location AMC, Department of Anesthesiology - L.E.I.C.A, Cardiovascular Science, Meibergdreef 11, 1105 AZ Amsterdam, the Netherlands. Electronic address: [email protected].
  • 2. Amsterdam, University Medical Centers, location AMC, Department of Anesthesiology - L.E.I.C.A, Cardiovascular Science, Meibergdreef 11, 1105 AZ Amsterdam, the Netherlands. Electronic address: [email protected].
  • 3. Amsterdam, University Medical Centers, location AMC, Department of Anesthesiology - L.E.I.C.A, Cardiovascular Science, Meibergdreef 11, 1105 AZ Amsterdam, the Netherlands. Electronic address: [email protected].
  • 4. Amsterdam, University Medical Centers, location AMC, Department of Anesthesiology - L.E.I.C.A, Cardiovascular Science, Meibergdreef 11, 1105 AZ Amsterdam, the Netherlands. Electronic address: [email protected].
  • 5. Amsterdam, University Medical Centers, location AMC, Department of Anesthesiology - L.E.I.C.A, Cardiovascular Science, Meibergdreef 11, 1105 AZ Amsterdam, the Netherlands. Electronic address: [email protected].
  • 6. Amsterdam, University Medical Centers, location AMC, Department of Anesthesiology - L.E.I.C.A, Cardiovascular Science, Meibergdreef 11, 1105 AZ Amsterdam, the Netherlands. Electronic address: [email protected].
  • 7. Amsterdam, University Medical Centers, location AMC, Department of Anesthesiology - L.E.I.C.A, Cardiovascular Science, Meibergdreef 11, 1105 AZ Amsterdam, the Netherlands. Electronic address: [email protected].
PMID: 36623448 DOI: 10.1016/j.biopha.2023.114228
Abstract

Background: 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.

Methods: 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.

Results: 10 µM CANA significantly reduced TNF-α related ROS generation, IL-1β production and NLRP3 expression (P all <0.05), but NAC did not alter the inflammasome activation (P > 0.05). CANA and BAPTA both prevented intracellular CA2+ increase in cells exposed to TNF-α (P both <0.05). Moreover, BAPTA and PD 98059 significantly reduced the TNF-α triggered IL-1β production as well as NLRP3 and pro-caspase-1 expression (P all <0.05).

Conclusion: CANA suppresses inflammasome activation by inhibition of (1) intracellular CA2+ and (2) ERK1/2 phosphorylation, but not by ROS reduction.

Keywords
Canagliflozin (CANA); Extracellular signal regulated kinase (ERK) 1/2 phosphorylation; Interleukin (IL)− 1β; Intracellular calcium (Ca(2+)); Nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome; Reactive oxygen species (ROS).
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