Effects of the (Pro)renin Receptor on Cardiac Remodeling and Function in a Rat Alcoholic Cardiomyopathy Model via the PRR-ERK1/2-NOX4 Pathway

Abstract

Alcoholic cardiomyopathy (ACM) caused by alcohol consumption manifests mainly as by maladaptive myocardial function, which eventually leads to heart failure and causes serious public health problems. The (pro)renin receptor (PRR) is an important member of the local tissue renin-angiotensin system and plays a vital role in many cardiovascular diseases. However, the mechanism responsible for the effects of PRR on ACM remains unclear. The purpose of this study was to determine the role of PRR in myocardial fibrosis and the deterioration of cardiac function in alcoholic cardiomyopathy. Wistar rats were fed a liquid diet containing 9% v/v alcohol to establish an alcoholic cardiomyopathy model. Eight weeks later, rats were injected with 1×109v.g./100 μl of recombinant adenovirus containing EGFP (scramble-shRNA), PRR, and PRR-shRNA via the tail vein. Cardiac function was assessed by echocardiography. Cardiac histopathology was measured by Masson’s trichrome staining, immunohistochemical staining, and dihydroethidium staining. In addition, cardiac fibroblasts (CFs) were cultured to evaluate the effects of alcohol stimulation on the production of the extracellular matrix and their underlying mechanisms. Our results indicated that overexpression of PRR in rats with alcoholic cardiomyopathy exacerbates myocardial oxidative stress and myocardial fibrosis. Silencing of PRR expression with short hairpin RNA (shRNA) technology reversed the myocardial damage mediated by PRR. Additionally, PRR activated phosphorylation of ERK1/2 and increased NOX4-derived reactive oxygen species and collagen expression in CFs with alcohol stimulation. Administration of the ERK kinase inhibitor (PD98059) significantly reduced NOX4 protein expression and collagen production, which indicated that PRR increases collagen production primarily through the PRR-ERK1/2-NOX4 pathway in CFs. In conclusion, our study demonstrated that PRR induces myocardial fibrosis and deteriorates cardiac function through ROS from the PRR-ERK1/2-NOX4 pathway during ACM development.

Activated ROCK/Akt/eNOS and ET-1/ERK pathways in 5-fluorouracil-induced cardiotoxicity: modulation by simvastatin

“…The exact cause of ROS overproduction was not clear; however, several pathways crosstalk increased ROS production. The 5-FU-associated ET-1 elevation activates Nox 71,72 , and in a feedforward signal, ROS stimulate ET-1 via the MAPK/ERK1/2-dependent pathway to further mediate a negative impact on ECs 73 . Moreover, ROS activate other molecules, such as COX isoforms that also contribute to endothelial dysfunction 73 ; all of which are facts that support the current findings.…”

Angiotensin II promotes podocyte injury by activating Arf6-Erk1/2-Nox4 signaling pathway

“…It has been reported that extracellular signal-regulated kinases (Erk1/2) play a critical role in regulation of Nox4 expression [19]. We then explored the activation level of Erk1/2 at Thr202 and Tyr204 following 1 μM of Ang II treatment.…”

(Pro)renin Receptor is Involved in Myocardial Damage in Alcoholic Cardiomyopathy

“…We constructed 3 short hairpin RNA (shRNA) fragments targeting PRR (Cao et al., ) which were encapsulated with recombinant adenovirus to form an adenovirus‐coated PRRshRNA fragment by Gene Pharma as follows:…”

“…Sirius Red staining was performed to evaluate myocardial interstitial fibrosis. Immunohistochemical staining was performed as previously described (Cao et al., ). Serial sections were incubated overnight at 4°C with primary antibodies against rat collagen I (1:250, Abcam, ab34710), fibronectin (1:200, Abcam, ab32419), nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 2 (NOX2, 1:200, Proteintech, China, 19013‐1‐AP), 3‐nitrotyrosine (3‐NT, 1:200, Abcam, ab61392), 4‐hydroxynonenal (4‐HNE, 1:100, Bioss, Beijing, China, bs‐6313R), IL‐1 β (1:500, Abcam, ab8320), and IL‐6 (1:500, Abcam, ab6672).…”