Smooth Muscle Cells of Dystrophic (mdx) Mice Are More Susceptible to Hypoxia; The Protective Effect of Reducing Ca 2+ Influx.
Arkady UryashAlfredo MijaresEric EstèveJose A AdamsJose R LopezPublished in: Biomedicines (2023)
Duchenne muscular dystrophy (DMD) is an inherited muscular disorder caused by mutations in the dystrophin gene. DMD patients have hypoxemic events due to sleep-disordered breathing. We reported an anomalous regulation of resting intracellular Ca 2+ ([Ca 2+ ] i ) in vascular smooth muscle cells (VSMCs) from a mouse (mdx) model of DMD. We investigated the effect of hypoxia on [Ca 2+ ] i in isolated and quiescent VSMCs from C57BL/10SnJ (WT) and C57BL/10ScSn-Dmd (mdx) male mice. [Ca 2+ ] i was measured using Ca 2+ -selective microelectrodes under normoxic conditions (95% air, 5% CO 2 ) and after hypoxia (glucose-free solution aerated with 95% N2-5% CO 2 for 30 min). [Ca 2+ ] i in mdx VSMCs was significantly elevated compared to WT under normoxia. Hypoxia-induced [Ca 2+ ] i overload, which was significantly greater in mdx than in WT VSMCs. A low Ca 2+ solution caused a reduction in [Ca 2+ ] i and prevented [Ca 2+ ] i overload secondary to hypoxia. Nifedipine (10 µM), a Ca 2+ channel blocker, did not modify resting [Ca 2+ ] i in VSMCs but partially prevented the hypoxia-induced elevation of [Ca 2+ ] i in both genotypes. SAR7334 (1 µM), an antagonist of TRPC3 and TRPC6, reduced the basal and [Ca 2+ ] i overload caused by hypoxia. Cell viability, assessed by tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, was significantly reduced in mdx compared to WT VSMCs. Pretreatment with SAR7341 increases cell viability in normoxic mdx ( p < 0.001) and during hypoxia in WT and mdx VSMCs. These results provide evidence that the lack of dystrophin makes VSMCs more susceptible to hypoxia-induced [Ca 2+ ] i overload, which appears to be mediated by increased Ca 2+ entry through L-type Ca 2+ and TRPC channels.
Keyphrases
- duchenne muscular dystrophy
- vascular smooth muscle cells
- protein kinase
- endothelial cells
- muscular dystrophy
- angiotensin ii
- type diabetes
- adipose tissue
- chronic kidney disease
- weight loss
- newly diagnosed
- single molecule
- copy number
- extracorporeal membrane oxygenation
- body composition
- high intensity
- prognostic factors
- atomic force microscopy