Induction of MicroRNA-21 with Exogenous Hydrogen Sulfide Attenuates Myocardial Ischemic and Inflammatory Injury in Mice
Background—Maintaining physiological levels of hydrogen sulfide (H2S) during ischemia is necessary to limit injury to the heart. Due to the anti-inflammatory effects of H2S, we proposed that the H2S donor, Na2S, would attenuate myocardial injury through upregulation of 'protective' microRNA (miR)-21 and suppression of the inflammasome, a macromolecular structure that amplifies inflammation and mediates further injury.
Methods and Results—Na2S-induced miR-21 expression was measured by qPCR in adult primary rat cardiomyocytes and in the mouse heart. We measured inflammasome formation and activity in cardiomyocytes challenged with lipopolysaccharide (LPS) and adenosine-tri-phosphate (ATP) or simulated ischemia/reoxygenation; and in the heart following regional myocardial ischemia/reperfusion (I/R), in the presence or absence of Na2S. To assess the direct anti-inflammatory effects of H2S in vivo, we utilized a peritonitis model by way of intraperitoneal injection of zymosan A. Na2S attenuated inflammasome formation and activity - measured by counting cytoplasmic aggregates of the scaffold protein Apoptosis Speck-like protein containing a Caspase-recruitment domain (ASC; -57%) and caspase-1 activity (-50%) in isolated cardiomyocytes and in the mouse heart (all P<0.05). Na2S also inhibited apoptosis (-38%) and necrosis (-43%) in cardiomyocytes in vitro and reduced myocardial infarct size (-63%) following I/R injury in vivo (all P<0.05). These protective effects were absent in cells treated with antagomiR-21 and in miR-21 KO mice. Na2S also limited the severity of inflammasome-dependent inflammation in the model of peritonitis (P<0.05) in wild-type but not in miR-21 KO mice.
Conclusions—Na2S induces cardioprotective effects through miR-21-dependent attenuation of ischemic and inflammatory injury in cardiomyocytes.
- Received September 6, 2013.
- Revision received March 14, 2014.
- Accepted April 3, 2014.