Abstract
Cardioprotective potential of anthocyanin rich red cabbage extract (ARCE) was assessed in H2O2 treated rat neonatal cardiomyoblasts (H9c2 cells) and isoproterenol (ISO) induced rodent model of myocardial infarction. H2O2 treated H9c2 cells recorded cytotoxicity (48–50%) and apoptosis (57.3%), the same were reduced in presence of ARCE (7–10% & 12.3% respectively). Rats pretreated with ARCE for 30 days followed by ISO treatment recorded favourable heart: body weight ratio as compared to ISO treated group. Also, the mRNA levels of enzymatic antioxidants (sod and catalase) and apoptotic genes (bax and bcl-2) in ARCE+ISO treated group were similar to the control group suggesting that ARCE pretreatment prevents ISO induced depletion of enzymatic antioxidants and apoptosis. Histoarchitecture of ventricular tissue of ISO treated group was marked by infracted areas (10%) and derangement of myocardium whereas, ARCE+ISO treated group (4.5%) recorded results comparable to control (0%). ARCE+ISO treated group accounted for upregulation of caveolin-3 and SERCA2a expression as compared to the ISO treated group implying towards ARCE mediated reduction in membrane damage and calcium imbalance. Molecular docking scores and LigPlot analysis of cyanidin-3-glucoside (-8.7 Kcal/mol) and delphinidin-3-glucoside (-8.5 Kcal/mol) showed stable hydrophobic and electrostatic interactions with β1 adrenergic receptor. Overall this study elucidates the mechanism of ARCE mediated prevention of experimentally induced myocardial damage.
Highlights
Cardiovascular diseases (CVDs) cause significant morbidity and continue to remain the leading cause of death globally
Annexin V-Alexa 488, Propidium Iodide (PI), TRIzol reagent, DreamTaq Green master mix and SYBR select master mix were procured from Invitrogen (CA, USA). iScript cDNA synthesis kit was purchased from Bio-Rad (CA, USA)
Thin Layer Chromatography (TLC) of anthocyanin rich red cabbage extract (ARCE) revealed two bands with Rf value 0.26 and 0.31 respectively. These values were in the Rf value range of 0.2–0.35 and which indicate presence of monoglucosides delphinidin-3-glucoside and cyanidin-3-glucoside [28] (Fig 1A)
Summary
Cardiovascular diseases (CVDs) cause significant morbidity and continue to remain the leading cause of death globally. Myocardial infarction (MI) is the most prevalent type of CVD wherein loss of cardiomyocytes due to apoptosis is at the epicentre of its pathogenesis [1]. Reactive oxygen species (ROS) mediated depletion of intracellular antioxidants, lipid peroxidation, modification of structural proteins and DNA damage precede apoptosis. Cardio-protective potential of Brassica oleracea L. reduction of intracellular ROS is one of the key targets of research to regulate apoptotic cascade in cardiomyocytes [2]. Rat cardiomyocytes when treated with H2O2, undergo cellular damage due to production of free radicals wherein, the sequences of events are similar to an oxidatively stressed myocardium. H2O2 induced oxidative stress and ISO induced myocardial infarction models are used to assess cardioprotective potential of the test compounds [5, 6]
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