2-oxothiazolidine-4-carboxylic acid inhibits vascular calcification via induction of glutathione synthesis

Jessal Patel, Lucie E Bourne, Shori Thakur, Kenneth Farrington, Diana Gorog, Isabel R Orriss, Anwar Baydoun

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Abstract

Arterial medial calcification (AMC), the deposition of hydroxyapatite in the medial layer of the arteries, is a known risk factor for cardiovascular events. Oxidative stress is a known inducer of AMC and endogenous antioxidants, such as glutathione (GSH), may prevent calcification. GSH synthesis, however, can be limited by cysteine levels. Therefore, we assessed the effects of the cysteine prodrug 2‐oxothiazolidine‐4‐carboxylic acid (OTC), on vascular smooth muscle cell (VSMC) calcification to ascertain its therapeutic potential. Human aortic VSMCs were cultured in basal or mineralising medium (1 mM calcium chloride/sodium phosphate) and treated with OTC (1–5 mM) for 7 days. Cell‐based assays and western blot analysis were performed to assess cell differentiation and function. OTC inhibited calcification ≤90%, which was associated with increased ectonucleotide pyrophosphatase/phosphodiesterase activity, and reduced apoptosis. In calcifying cells, OTC downregulated protein expression of osteoblast markers (Runt‐related transcription factor 2 and osteopontin), while maintaining expression of VSMC markers (smooth muscle protein 22α and α‐smooth muscle actin). GSH levels were significantly reduced by 90% in VSMCs cultured in calcifying conditions, which was associated with declines in expression of gamma‐glutamylcysteine synthetase and GSH synthetase. Treatment of calcifying cells with OTC blocked the reduction in expression of both enzymes and prevented the decline in GSH. This study shows OTC to be a potent and effective inhibitor of in vitro VSMC calcification. It appears to maintain GSH synthesis which may, in turn, prevent apoptosis and VSMCs gaining osteoblast‐like characteristics. These findings may be of clinical relevance and raise the possibility that treatment with OTC could benefit patients susceptible to AMC.
Original languageEnglish
Article number30036
JournalJournal of Cellular Physiology
Volume2020
Early online date12 Sept 2020
DOIs
Publication statusE-pub ahead of print - 12 Sept 2020

Keywords

  • 2-oxothiazolidine-4-carboxylic acid
  • arterial medial calcification
  • glutathione
  • reactive oxygen species
  • vascular smooth muscle cells

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