Cardiotonic Pill Reduces Myocardial Ischemia-Reperfusion Injury via Increasing EET Concentrations in Rats

Abstract

Accumulating data suggest that epoxyeicosatrienoic acids (EETs) and 20-hydroxyeicosatetraenoic acid, both cytochrome P450 (P450) enzyme metabolites of arachidonic acid (AA), play important roles in cardiovascular diseases. For many years, the cardiotonic pill (CP), an herbal preparation derived from Salviae Miltiorrhizae Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Borneolum Syntheticum, has been widely used in China for the treatment of coronary artery disease. However, its pharmacological mechanism has not been well elucidated. The purpose of this study was to investigate the chronic effects of the CP on myocardial ischemia-reperfusion injury (MIRI) and AA P450 enzyme metabolism in rats (in vivo) and H9c2 cells (in vitro). The results showed that CP dose dependently (10, 20, and 40 mg/kg/d; 7 days) mitigated MIRI in rats. The plasma concentrations of EETs in CP-treated ischemia-reperfusion (I/R) rats (40 mg/kg/d; 7 days) were significantly higher (P < 0.05) than those in controls. Cardiac Cyp1b1, Cyp2b1, Cyp2e1, Cyp2j3, and Cyp4f6 were significantly induced (P < 0.05); CYP2J and CYP2C11 proteins were upregulated (P < 0.05); and AA-epoxygenases activity was significantly increased (P < 0.05) after CP (40 mg/kg/d; 7 days) administration in rats. In H9c2 cells, the CP also increased (P < 0.05) the EET concentrations and showed protection in hypoxia-reoxygenation (H/R) cells. However, an antagonist of EETs, 14,15-epoxyeicosa-5(Z)-enoic acid, displayed a dose-dependent depression of the CP's protective effects in H/R cells. In conclusion, upregulation of cardiac epoxygenases after multiple doses of the CP-leading to elevated concentrations of cardioprotective EETs after myocardial I/R-may be the underlying mechanism, at least in part, for the CP's cardioprotective effect in rats.

Genetic Deletion or Pharmacological Inhibition of Soluble Epoxide Hydrolase Ameliorates Cardiac Ischemia/Reperfusion Injury by Attenuating NLRP3 Inflammasome Activation

“…It is recognized the current study was limited by not directly assessing changes in the levels of epoxylipids (EETs, EDPs, EpOMEs) as well as the diol metabolites (DiHOMEs) in response to IR injury. However, it has been shown that cardiac ischemic insults decrease levels of the cardioprotective epoxylipids EETs and EDPs and cardiotoxic DiHOMEs accumulate aggravating the injury [75,118]. Therefore, future characterization of how IR injury alters the levels of these metabolites will provide more insight into the complex responses and pathobiology.…”

Absolute protein assay for the simultaneous quantification of two epoxide hydrolases in rats by mass spectrometry–based targeted proteomics

“…To verify the results of protein quantification, an enzyme activity investigation was conducted. Various compounds, such as epoxyeicosatrienoic acid ethanolamides [5], trans ‐stilbene oxide [10], 4‐nitrophenyl‐trans‐2,3‐epoxy‐3‐phenyl propyl carbonate [9], and EETs [24], have been used as the substrate of the epoxide hydrolases. In this study, the metabolic activity of total epoxide hydrolase in heart, liver, and kidney S9 samples was determined by the ring‐open hydrolysis reaction of 14,15‐EET.…”

“…To obtain the total epoxide hydrolase activity of mEH and sEH in S9 samples, we performed an established in vitro study as described previously with a minor modification [24]. In brief, the S9 fraction was diluted to 0.012 mg/mL with PBS (0.1 M, pH 7.4).…”

“…The residue was dissolved in 120 µL of dehydrated alcohol and vortexed for 1 min. The tubes were then centrifuged at 12 000 × g for 10 min, and 5 µL of the supernatant was analyzed using an LC‐MS/MS method as described elsewhere [24].…”

MicroRNA-1 and Circulating Microvesicles Mediate the Protective Effects of Dantonic in Acute Myocardial Infarction Rat Models

“…The clinical value of Dantonic in the treatment of chronic stable angina pectoris was confirmed again by Phase III clinical trial of FDA. Previous studies showed that Dantonic treatment could significantly reduce the ischemia/reperfusion- (I/R-) and LAD induced myocardial damage and apoptosis, myocardial fibrosis, microcirculatory disturbance ( Zhao et al, 2010 ; Wei et al, 2013 ; Yang et al, 2013 ), oxidative stress, and inflammatory response ( Horie et al, 2005 ; Lee et al, 2013 ; Jun et al, 2014 ; Ren-an et al, 2014 ; Chen et al, 2016 ; Xu et al, 2016 ).…”