Paeonol Pretreatment Attenuates Anoxia-Reoxygenation Induced Injury in Cardiac Myocytes <i>via</i> a BRCA1 Dependent Pathway

Abstract

Breast cancer type 1 sensitive protein (BRCA1) is a well-known tumor suppressor and its role in oxidative stress has been confirmed. The purpose of this study is to evaluate whether paeonol has a protective effect on myocardial hypoxia-reoxygenation (A/R) injury, and to explore H9C2 cells through a mechanismdependent pathway mediated by BRCA1. H9C2 cells were pretreated with paeonol (10 µM) for 18 h before hypoxia was induced to establish a cell model of myocardial ischemia/reperfusion (I/R) injury. Use commercial kits to detect antioxidant indicators, including relative oxygen content (ROS) levels, total antioxidant capacity (T-AOC), superoxide dismutase (SOD), lactate dehydrogenase (LDH) activity, and creatine kinase (CK-MB) and nuclear factor-kappaB (NF-κB) activity. The cell viability was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction method. Real-time fluorescent quantitative PCR was used to detect BRCA1 mRNA and protein levels. The expression levels of BRCA1, NLRP3 and ACS were determined by Western blotting. In addition, the release of interleukin (IL)-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) was also evaluated by an enzyme-linked immunosorbent assay (ELISA) kit. The results showed that paeonol (10 µM) can significantly improve the hypoxic A/R damage of H9C2 cells, and the BRCA1 expression of H9C2 cells pretreated with paeonol was significantly increased before A/R damage was induced. BRCA1 is widely known in breast and ovarian cancer. Our data proves that the down-regulation of BRCA1 participates in the decrease of cell viability and the decrease of CK-MB and LDH activities, and protects cells by inhibiting the production of ROS and the activation of Nod-like receptor protein 3 (NLRP3) inflammasomes and NF-κB. In conclusion, paeonol significantly improved the A/R damage of H9C2 cells induced by hypoxia through the BRCA1/ROS-regulated NLRP3 inflammasome/IL-1β and NF-κB/TNF-α/IL-6 pathways. It may be a potential drug against myocardial I/R injury.

Active Ingredient Paeonol of Jijiu Huiyang Decoction Alleviates Isoproterenol-Induced Chronic Heart Failure via the GSK3A/PPARα Pathway

“…After that, the total RNA underwent reverse transcription and quantification on a Real-Time PCR system (7500, ThermoFisher, USA) using the One-Step SYBR Green qRT-PCR Kit (BL643B, Biosharp, China). Under the adoption of the 2 − ΔΔ CT method, B-type natriuretic peptide (BNP), atrial natriuretic peptide (ANP), major histocompatibility complex-B (B-MHC), GSK3A, CD36, PDK4 (Pyruvate dehydrogenase kinase 4), and Cpt1b (carnitine palmitoyltransferase 1B) expressions were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) [ 12 ]. Supplementary Table 3 lists the primer sequences.…”

Promising Therapeutic Candidate for Myocardial Ischemia/Reperfusion Injury: What Are the Possible Mechanisms and Roles of Phytochemicals?

“…NF-κB interacts with the nucleus by regulating more than 200 genes, among which some produce inflammatory cytokines, which ultimately lead to excessive inflammation. H9C2 cells, which are damaged by hypoxia through BRCA1/ROS-regulated NLRP3 inflammasome/IL-1β and NF-κB/TNF-α/IL-6 pathways, can be improved by Paeonol ( 44 ). When intercellular macromolecular proteins aggregate, they are called inflammasomes, which promote the maturation of inflammatory cytokines ( 45 ).…”

“…Pretreating with paeonol can significantly improve the hypoxia-reoxygenation (H/R) damage and the BRCA1 expression of H9C2 cells through the BRCA1/ROS-regulated NLRP3 inflammasome/IL-1β and NF-κB/TNF-α/IL-6 pathways. It may be a candidate drug for treating myocardial I/R injury ( 44 ).…”

“…Activated NF-κB prevents ischemic injury and inhibits both inflammation and apoptosis ( 304 ). Paeonol significantly alleviates hypoxia and attenuates I/R injury in H9C2 cells through the BRCA1/ROS-regulated NF-κB/TNF-α/IL-6 pathways and NLRP3 inflammasome ( 44 ). Puerarin exerts a similar effect by suppressing NF-κB and upregulating VEGFA/Ang-1 in diabetic rats with myocardial I/R injury ( 94 ).…”

Cardioprotective Effect of Paeonol on Chronic Heart Failure Induced by Doxorubicin via Regulating the miR-21-5p/S-Phase Kinase-Associated Protein 2 Axis

“…Paeonol has currently been proven to have a cardioprotective effect on epirubicin-induced heart injury ( 18 ). Additionally, recent studies further demonstrated that paeonol pretreatment attenuates hypoxia-reoxygenation-induced injury of cardiomyocytes via the breast cancer type 1 sensitive protein (BRCA1)-dependent pathway, and paeonol attenuates transverse aortic constriction-induced HF through the ERK1/2 signaling pathway ( 19 , 20 ). Also, paeonol suppresses vasculogenesis via regulating vascular smooth muscle phenotypic switching, and it promotes Opa1-mediated mitochondrial fusion by activating the CK2α-Stat3 pathway in diabetic cardiomyopathy ( 21 , 22 ).…”