Survival-Related Autophagic Activity Versus Procalcific Death in Cultured Aortic Valve Interstitial Cells Treated With Critical Normophosphatemic-Like Phosphate Concentrations

Overview

Journal J Histochem Cytochem

Publisher Sage Publications

Specialty Biochemistry

Date 2017 Jan 24

PMID 28112549

Citations 9

Authors

Antonella Bonetti

Alberto Della Mora

Magali Contin

Giorgia Gregoraci

Franco Tubaro

Maurizio Marchini

Fulvia Ortolani

Affiliations

Soon will be listed here.

Abstract

Valve dystrophic calcification is a common disorder affecting normophosphatemic subjects. Here, cultured aortic valve interstitial cells (AVICs) were treated 3 to 28 days with phosphate (Pi) concentrations spanning the normal range in humans (0.8, 1.3, and 2.0 mM) alone or supplemented with proinflammatory stimuli to assess possible priming of dystrophic-like calcification. Compared with controls, spectrophotometric analyses revealed marked increases in calcium amounts and alkaline phosphatase activity for 2.0-mM-Pi-containing cultures, with enhancing by proinflammatory mediators. Ultrastructurally, AVICs treated with low/middle Pi concentrations showed an enormous endoplasmic reticulum (ER) enclosing organelle debris, so apparently executing a survival-related atypical macroautophagocytosis, consistently with ultracytochemical demonstration of ER-associated acid phosphatase activity and decreases in autophagosomes and immunodetectable MAP1LC3. In contrast, AVICs cultured at 2.0-mM Pi underwent mineralization due to intracellular release and peripheral layering of phospholipid-rich material acting as hydroxyapatite nucleator, as revealed by Cuprolinic Blue and von Kossa ultracytochemical reactions. Lack of immunoblotted caspase-3 cleaved form indicated apoptosis absence for all cultures. In conclusion, fates of cultured AVICs were crucially driven by Pi concentration, suggesting that serum Pi levels just below the upper limit of normophosphatemia in humans may represent a critical watershed between macroautophagy-associated cell restoring and procalcific cell death.

Citing Articles

Ultrastructural and Immunohistochemical Detection of Hydroxyapatite Nucleating Role by rRNA and Nuclear Chromatin Derivatives in Aortic Valve Calcification: In Vitro and In Vivo Pro-Calcific Animal Models and Actual Calcific Disease in Humans.

Bonetti A, Contin M, Marchini M, Ortolani F Int J Mol Sci. 2023; 24(3).

PMID: 36768988 PMC: 9916520. DOI: 10.3390/ijms24032667.

Calcium-Dependent Cytosolic Phospholipase A2α as Key Factor in Calcification of Subdermally Implanted Aortic Valve Leaflets.

Bonetti A, Contin M, Tonon F, Marchini M, Ortolani F Int J Mol Sci. 2022; 23(4).

PMID: 35216105 PMC: 8877272. DOI: 10.3390/ijms23041988.

Self-eating and Heart: The Emerging Roles of Autophagy in Calcific Aortic Valve Disease.

Fan Y, Shao J, Wei S, Song C, Li Y, Jiang S Aging Dis. 2021; 12(5):1287-1303.

PMID: 34341709 PMC: 8279526. DOI: 10.14336/AD.2021.0101.

The Protective Effects of the Autophagic and Lysosomal Machinery in Vascular and Valvular Calcification: A Systematic Review.

Neutel C, Hendrickx J, Martinet W, De Meyer G, Guns P Int J Mol Sci. 2020; 21(23).

PMID: 33255685 PMC: 7728070. DOI: 10.3390/ijms21238933.

COX-2 Is Downregulated in Human Stenotic Aortic Valves and Its Inhibition Promotes Dystrophic Calcification.

Vieceli Dalla Sega F, Fortini F, Cimaglia P, Marracino L, Tonet E, Antonucci A Int J Mol Sci. 2020; 21(23).

PMID: 33255450 PMC: 7727817. DOI: 10.3390/ijms21238917.

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