Inorganic phosphate (Pi) modulates the expression of key regulatory proteins of the inorganic pyrophosphate (PPi) metabolism in TGF-β1-stimulated chondrocytes
Affiliations: Laboratoire de Physiopathologie, Pharmacologie et Ingénierie Articulaires, CNRS-Nancy University, Vandoeuvre-lès-Nancy, France
Note: [] Address for correspondence: Arnaud Bianchi and Frédéric Cailotto, Laboratoire de Physiopathologie, Pharmacologie et Ingénierie Articulaires (LPPIA), UMR 7561 CNRS-Nancy University, Vandoeuvre-lès-Nancy, France. Tel.: +33 3 83 68 39 50; Fax: +33 3 83 68 39 59; E-mails: frederic.cailotto@hotmail.fr, arnaud.bianchi@medecine.uhp-nancy.fr.
Abstract: The balance between extracellular inorganic phosphate (ePi) and extracellular inorganic pyrophosphate (ePPi) is controlled by four membrane proteins: the transporters ANK (exporting PPi outside the cells) and PiT-1 (importing ePi into the cells), and the enzymes PC-1 (generating ePPi from nucleotides) and Tissue Non-specific Alkaline Phosphatase (TNAP, hydrolyzing ePPi into ePi). TGF-β1 was shown to stimulate ANK and PC-1 expression in articular chondrocytes, and subsequent ePPi level, as well as to increase ePi uptake by inducing PiT-1 expression in a chondrogenic cell line. Thus, we investigated the ability of ePi to modulate the effect of TGF-β1 on the regulatory proteins of the ePi/ePPi balance in chondrocytes. In the pathophysiological range of 0.01–1 mM, ePi was inactive by itself but potentiated the stimulatory effects of TGF-β1 on ANK, PC-1 or PiT-1 mRNA (RT-qPCR) and protein (Western blot) levels. PC-1 activity was also increased by TGF-β1 and further potentiated by ePi supplementation. TNAP mRNA and activity became undetectable in response to TGF-β1. These data suggest that ePi could increase ePPi level by changing the control of ANK and PC-1 expression by TGF-β1, further highlighting an adaptative regulation of the Pi/PPi balance to prevent basic calcium phosphate deposition into the joints.
