From the, Department of Cardiology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.
The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital of Shandong University, Jinan, China.
Alcohol Clin Exp Res. 2019 Nov;43(11):2344-2353. doi: 10.1111/acer.14188. Epub 2019 Oct 8.
(Pro)renin receptor (PRR), a novel member of the renin-angiotensin system, participates in various cardiovascular diseases. However, the role of PRR in alcoholic cardiomyopathy (ACM), which is caused by alcohol intake and manifests as myocardial damage and cardiac dysfunction, remains unclear.
PRR gene silencing was achieved by transfecting recombinant adenovirus expressing anti-PRR short hairpin RNA (PRR-shRNA). In vitro, primary rat cardiac fibroblasts (CFs) were cultured with the stimulation of alcohol (200 mM), with or without PRR-shRNA and PD98059. Immunofluorescence, RT-PCR, and Western blot were used to measure the protein and messenger (mRNA) expression of PRR, fibrotic factors, and members of related signaling pathways. In vivo, Wistar rats were fed a diet containing 9% (v/v) alcohol or a normal diet for 3 months, with or without PRR-shRNA. Sirius Red staining, immunohistochemical staining, and toluidine blue staining were used to evaluate myocardial fibrosis, oxidative stress, and inflammation response.
Alcohol markedly increased PRR mRNA and protein expression in a time- and concentration-dependent manner in CFs. The increased expression of fibrotic factors induced by alcohol was prevented by PRR-shRNA and PD98059. Moreover, PRR-shRNA decreased the phosphorylation of extracellular regulated protein kinases (ERK) 1/2 in CFs. Furthermore, PRR-shRNA decreased cardiac fibrosis, reduced oxidative stress, and alleviated inflammation response in the myocardial tissue.
Our results show that PRR-ERK1/2 signaling was involved in the development of ACM and that PRR could be a new target for the treatment of ACM.
（前）肾素受体（PRR）是肾素-血管紧张素系统的一个新成员，参与多种心血管疾病。然而，PRR 在由酒精摄入引起的心肌损伤和心功能障碍的酒精性心肌病（ACM）中的作用尚不清楚。
通过转染表达抗 PRR 短发夹 RNA（PRR-shRNA）的重组腺病毒实现 PRR 基因沉默。在体外，用酒精（200mM）刺激原代大鼠心肌成纤维细胞（CFs），同时用或不用 PRR-shRNA 和 PD98059。免疫荧光、RT-PCR 和 Western blot 用于测量 PRR、纤维化因子和相关信号通路成员的蛋白和信使（mRNA）表达。在体内，用含 9%（v/v）酒精的饮食或正常饮食喂养 Wistar 大鼠 3 个月，同时用或不用 PRR-shRNA。天狼猩红染色、免疫组织化学染色和甲苯胺蓝染色用于评估心肌纤维化、氧化应激和炎症反应。
酒精在 CFs 中以时间和浓度依赖的方式显著增加 PRR mRNA 和蛋白的表达。PRR-shRNA 和 PD98059 可阻止酒精诱导的纤维化因子表达增加。此外，PRR-shRNA 降低了 CFs 中细胞外调节蛋白激酶（ERK）1/2 的磷酸化。此外，PRR-shRNA 减少了心肌组织中的心脏纤维化、降低了氧化应激并减轻了炎症反应。
我们的结果表明，PRR-ERK1/2 信号通路参与了 ACM 的发生，PRR 可能成为 ACM 治疗的新靶点。
