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Articles in PresS, published online ahead of print November 20, 2001
Am J Physiol Renal Physiol, 10.1152/ajprenal.00195.2001
Submitted on June 26, 2001
Accepted on November 16, 2001
1 Anesthesiology, Columbia University, New York, NY, USA
* To whom correspondence should be addressed. E-mail: tl128{at}columbia.edu.
We have recently demonstrated protection against renal ischemic-reperfusion (IR) injury in vivo by A1 and A2a adenosine receptor (AR) modulations. To further elucidate the signaling cascades of AR induced cytoprotection against reperfusion/oxidant-mediated injury, immortalized human proximal tubule (HK-2) cells were treated with hydrogen peroxide (H2O2). H2O2 caused dose and time dependent HK-2 cell death measured by LDH release and trypan blue dye uptake. Adenosine protected against H2O2-induced HK-2 cell death via A1 and A2a AR activation. A1 AR mediated protection involves pertussis toxin sensitive G-proteins and PKC, whereas A2a AR mediated protection involves PKA activation via cAMP and activation of CREB. Moreover, PKA activators (forskolin and Sp-CAMPS) also protected HK-2 cells against H2O2 injury. De novo gene transcription and protein synthesis are required for both A1 and A2a AR mediated cytoprotection as actinomycin D and cycloheximide, respectively, blocked cytoprotection. Chronic treatments with a non-selective AR agonist abolished the protection by adenosine. Moreover, chronic treatments with a non-selective AR antagonist increased the endogenous tolerance of HK-2 cells against H2O2. We conclude that A1 and A2a AR activation protect HK-2 cells against H2O2 induced injury via distinct signaling pathways requiring new gene transcription and new protein synthesis.
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