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Submitted on November 24, 2004
Accepted on March 15, 2005
Department of Internal Medicine University of Pisa, I-56100 Pisa, Italy; Department of Experimental and Diagnostic Medicine, Section of General Pathology, Interdisciplinary Center for the Study of Inflammation (ICSI) University of Ferrara, I-44100 Ferrara, Italy
* To whom correspondence should be addressed. E-mail: a.solini{at}med.unipi.it.
We investigated the presence of P2 receptors (P2Rs) in human thyrocytes and their possible involvement in the modulation of cytokine release. P2Rs expression was assessed by RT-PCR and, when possible, by immunoblotting. Human primary thyrocytes express the mRNA for the following P2X and P2Y subtypes: P2X3, P2X5, P2X6, P2X7 and P2Y1, P2Y2, P2Y4, and P2Y11. Stimulation with extracellular nucleotides of fura-2-loaded thyrocytes triggered an intracellular Ca2+ signal, suggesting expression of functional receptors. Thyrocytes spontaneously released the proinflammatory cytokine interleukin-6 (IL-6). The ATP-hydrolyzing enzyme apyrase reduced basal IL-6 release, while extracellular ATP dose-dependently increased IL-6 secretion. UTP was also an effective stimulus, while benzoyl-ATP was ineffective, suggesting a P2Y rather than P2X-modulated response. Finally, TSH reduced both the intracellular Ca2+ ([Ca2+]i) rise and IL-6 release triggered by P2Rs stimulation. In conclusion, we provide functional, pharmacological, and biochemical evidence that human primary thyrocytes express P2YR and P2XR subtypes, coupled to increases in ([Ca2+]i) and secretion of IL-6. P2R-dependent modulation of IL-6 release from human thyrocytes suggests a novel mechanism whereby an inflammatory and/or immune-mediated damage can be initiated and amplified in the thyroid.
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