Evaluation of antioxidant drugs for the treatment of ochronotic alkaptonuria in an in vitro human cell model.

Abstract

Alkaptonuria (AKU) is a rare autosomal recessive disease, associated with deficiency of homogentisate 1,2-dioxygenase activity in the liver. This leads to an accumulation of homogentisic acid (HGA) and its oxidized derivatives in polymerized form in connective tissues especially in joints. Currently, AKU lacks an appropriate therapy. Hence, we propose a new treatment for AKU using the antioxidant N-acetylcysteine (NAC) administered in combinations with ascorbic acid (ASC) since it has been proven that NAC counteracts the side-effects of ASC. We established an in vitro cell model using human articular primary chondrocytes challenged with an excess of HGA (0.33 mM). We used this experimental model to undertake pre-clinical testing of potential antioxidative therapies for AKU, evaluating apoptosis, viability, proliferation, and metabolism of chondrocytes exposed to HGA and treated with NAC and ASC administered alone or in combination addition of both. NAC decreased apoptosis induced in chondrocytes by HGA, increased chondrocyte growth reduced by HGA, and partially restored proteoglycan release inhibited by HGA. A significantly improvement in efficacy was found with combined addition of the two antioxidants in comparison with NAC and ASC alone. Our novel in vitro AKU model allowed us to demonstrate the efficacy of the co-administration of NAC and ASC to counteract the negative effects of HGA for the treatment of ochronotic arthropathy.

• Authors: Laura Tinti, Adriano Spreafico, Daniela Braconi, Lia Millucci, Giulia Bernardini, Federico Chellini, Giovanni Cavallo, Enrico Selvi, Mauro Galeazzi, Roberto Marcolongo, James A Gallagher, Annalisa Santucci
• Journal: Journal of cellular physiology (J Cell Physiol) Vol. 225 Issue 1 Pg. 84-91 (Oct 2010) ISSN: 1097-4652 [Electronic] United States
• PMID: 20648626 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: (c) 2010 Wiley-Liss, Inc.

Chemical References

• Antioxidants
• Homogentisate 1,2-Dioxygenase
• Homogentisic Acid
• Ascorbic Acid
• Acetylcysteine

Topics

• Acetylcysteine (pharmacology, therapeutic use)
• Alkaptonuria (drug therapy, enzymology, pathology)
• Antioxidants (pharmacology, therapeutic use)
• Apoptosis (drug effects)
• Ascorbic Acid (pharmacology, therapeutic use)
• Cartilage, Articular (cytology)
• Cell Proliferation (drug effects)
• Cell Survival (drug effects)
• Cells, Cultured
• Chondrocytes (drug effects, metabolism)
• Homogentisate 1,2-Dioxygenase (genetics, metabolism)
• Homogentisic Acid (metabolism)
• Humans
• Ochronosis (drug therapy, enzymology, pathology)
• Protein Carbonylation