Blockade of the natriuretic peptide receptor guanylyl cyclase-A inhibits NF-κB activation and alleviates myocardial ischemia/reperfusion injury
Takehiko Izumi, … , Seibu Mochizuki, Kazuwa Nakao
Takehiko Izumi, … , Seibu Mochizuki, Kazuwa Nakao
Published July 15, 2001
Citation Information: J Clin Invest. 2001;108(2):203-213. https://doi.org/10.1172/JCI12088.
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Article

Blockade of the natriuretic peptide receptor guanylyl cyclase-A inhibits NF-κB activation and alleviates myocardial ischemia/reperfusion injury

Abstract

Acute myocardial infarction (AMI) remains the leading cause of death in developed countries. Although reperfusion of coronary arteries reduces mortality, it is associated with tissue injury. Endothelial P-selectin–mediated infiltration of neutrophils plays a key role in reperfusion injury. However, the mechanism of the P-selectin induction is not known. Here we show that infarct size after ischemia/reperfusion was significantly smaller in mice lacking guanylyl cyclase-A (GC-A), a natriuretic peptide receptor. The decrease was accompanied by decreases in neutrophil infiltration in coronary endothelial P-selectin expression. Pretreatment with HS-142-1, a GC-A antagonist, also decreased infarct size and P-selectin induction in wild-type mice. In cultured endothelial cells, activation of GC-A augmented H2O2-induced P-selectin expression. Furthermore, ischemia/reperfusion–induced activation of NF-κB, a transcription factor that is known to promote P-selectin expression, is suppressed in GC-A–deficient mice. These results suggest that inhibition of GC-A alleviates ischemia/reperfusion injury through suppression of NF-κB–mediated P-selectin induction. This novel, GC-A–mediated mechanism of ischemia/reperfusion injury may provide the basis for applying GC-A blockade in the clinical treatment of reperfusion injury.

Authors

Takehiko Izumi, Yoshihiko Saito, Ichiro Kishimoto, Masaki Harada, Koichiro Kuwahara, Ichiro Hamanaka, Nobuki Takahashi, Rika Kawakami, Yuhao Li, Genzo Takemura, Hisayoshi Fujiwara, David L. Garbers, Seibu Mochizuki, Kazuwa Nakao

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Figure 7

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NF-κB activation, IκBα phosphorylation, and IκBα degradation in ANP- and...
NF-κB activation, IκBα phosphorylation, and IκBα degradation in ANP- and/or H2O2-treated HUVECs. (a) EMSA of NF-κB in control cells (lane 1), ANP-treated cells (lane 2), H2O2-treated cells (lane 3), ANP with H2O2-treated cells (lane 4), ANP and HS with H2O2-treated cells (lane 5). An arrow in right side indicates shifted band. (b) Semiquantitative analysis of the binding of activated NF-κB to DNA in HUVECs. Level of NF-κB activation was significantly higher in H2O2-treated cells (H)compared with control cells (N) and was further increased in ANP with H2O2-treated cells (H+A) compared with H2O2-treated cells. This effect was abolished by treatment with HS (H+A+HS). *P < 0.01 vs. control cells. #P < 0.01 vs. H2O2-treated cells. Western blot analysis of IκBα phosphorylation (c) and total IκBα protein expression (d) in HUVECs. Lane 1, control cells; lane 2, ANP-treated cells; lane 3, H2O2-treated cells; lane 4, H2O2 with ANP-treated cells; lane 5, H2O2 with ANP- and HS-treated cells. IκBα was not phosphorylated by ANP alone, but by H2O2, and IκBα phosphorylation was further strengthened by ANP with H2O2. This effect was abolished by HS (c). IκBα protein expression did not change among all groups (d). Densitometric scanning of IκBα phosphorylation (e) and IκBα protein expression (f). Phosphorylation of IκBα in H2O2-treated cells is significantly increased compared with control cells. It further increased significantly in H2O2 with ANP-treated cells compared with H2O2-treated cells (e). There is no significant change in IκBα protein expression among all groups (f). *P < 0.05 vs. control cells. #P < 0.01 vs. H2O2-treated cells.