Protective role of the novel hybrid 3,5-dipalmitoyl-nifedipine in a cardiomyoblast culture subjected to simulated ischemia/reperfusion Eduarda Santa-Helena a,b , Stefanie Teixeira b , Micheli Rosa de Castro a , Diego da Costa Cabrera c , Caroline Da Ros Montes D’Oca c , Marcelo G. Montes D’Oca c , Ana Paula S. Votto a,b , Luiz Eduardo Maia Nery a,b, * , 1 , Carla Amorim Neves Gonçalves a,b a Graduate Program in Physiological Sciences, Comparative Animal Physiology, Institute of Biological Sciences, Universidade Federal do Rio Grande, FURG, Rio Grande, RS, Brazil b Institute of Biological Sciences, Universidade Federal do Rio Grande, FURG, Rio Grande, RS, Brazil c Kolbe Organic Synthesis Laboratory, School of Chemistry and Food, FURG, Rio Grande, RS, Brazil A R T I C L E I N F O Article history: Received 20 December 2016 Received in revised form 22 May 2017 Accepted 22 May 2017 Keywords: Calcium blocker H9c2 Hypoxia Infarction Palmitic acid A B S T R A C T This work investigated the acute effects of the calcium channel blocker nifedipine and its new fatty hybrid derived from palmitic acid, 3,5-dipalmitoyl-nifedipine, compared to endocannabinoid ananda- mide during the process of inducing ischemia and reperfusion in cardiomyoblast H9c2 heart cells. The cardiomyoblasts were treated in 24 or 96-well plates (according to the test being performed) and maintaining the treatment until the end of hypoxia induction. The molecules were tested at concentrations of 10 and 100 mM, cells were treated 24 h after assembling the experimental plates and immediately before the I/R. Cell viability, apoptosis and necrosis, and generation of reactive oxygen species were evaluated. Nifedipine and 3,5-dipalmitoyl-nifedipine were used to assess radical scavenging potential and metal chelation. All tested molecules managed to reduce the levels of reactive oxygen species compared to the starvation + vehicle group. In in vitro assays, 3,5-dipalmitoyl-nifedipine showed more antioxidant activity than nifedipine. These results indicate the ability of this molecule to act as a powerful ROS scavenger. Cell viability was highest when cells were induced to I/R by both concentrations of anandamide and the higher concentration of DPN. These treatments also reduced cell death. Therefore, it was demonstrated that the process of hybridization of nifedipine with two palmitic acid chains assigns a greater cardioprotective effect to this molecule, thereby reducing the damage caused by hypoxia and reoxygenation in cardiomyoblast cultures. © 2017 Elsevier Masson SAS. All rights reserved. 1. Introduction Cardiovascular disease is a global public health problem. Damage after myocardial ischemia and reperfusion (I/R) is the leading cause of morbidity and mortality globally [1,2]. Hyperten- sive patients are most affected by I/R [3], stressing the importance of studies investigating drugs that attenuate blood pressure while concurrently preventing or mitigating I/R myocardial damage. Evidence indicates that several interrelated factors, such as the decrease in cellular ATP levels, the production of reactive oxygen species (ROS), the accumulation of hydrogen ions and the generation of reactive nitrogen species, contribute to the damage caused by I/R [4]. Oxidative stress contributes to the cascade of events leading to cell death: increased ROS production may modify the expression of various inflammatory mediators during cardiac injury [5]. During reoxygenation, the membrane integrity is compromised by the oxidation of the phospholipids which leads to uncontrolled ion permeability [6]. The ROS can still compromise the function of cardiac proteins, such as ion channels, calcium pumps and contractile proteins involved in the excitation mechanism of heart contraction [7]. Thus, the restoration of blood flow and the return of the oxygen supply generate ROS that trigger cell death through apoptosis and necrosis [6]. Substances that have anti-hypertensive and antioxidant action can bring benefits for the treatment of hypertensive patients prone to ischemia and reperfusion injury. According to Benzie and Tomlinson [8], antihypertensive drugs such as captopril, fosinopril, * Corresponding author at: Institute of Biological Sciences, Universidade Federal do Rio Grande, FURG, Av. Itália Km 8, CEP 96201-900, Rio Grande, RS, Brazil. E-mail address: famnery@terra.com.br (L.E.M. Nery). 1 Institute of Biological Sciences, Universidade Federal do Rio Grande, FURG, Rio Grande, RS, 96203-900, Brazil. http://dx.doi.org/10.1016/j.biopha.2017.05.091 0753-3322/© 2017 Elsevier Masson SAS. All rights reserved. Biomedicine & Pharmacotherapy 92 (2017) 356–364 Available online at ScienceDirect www.sciencedirect.com