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Cypagen 200

The performance-enhancing agents are commonly abused by skilled athletes. Among these agents, many have no proven merits and are linked to serious adverse effects . Androgenic anabolic steroids (AASs), such as Boldenone, are abused to enhance muscle mass, strength, and growth as well as to enhance athletic performance. Many countries have forbidden the use of AAS due to their adverse effects.

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The effect of anabolic androgenic steroids on the cardiovascular system is poorly understood. Increased production of free radicals is coupled to the pathophysiology of many alterations within the circulatory system. The only function of the enzyme family NADPH oxidases (NOXs) is the generation of reactive oxygen species (ROS). Therefore, this study investigated the beneficial role of grape seed proanthocyanidin extract (GSPE) in ameliorating cardiac toxicity induced by the anabolic steroid Boldenone in male rats through NOX inhibition and reduction in the expression of NOX2 and NOX4. This study was conducted on forty male rats which are divided into four groups (normal control, positive control or GSPE, Boldenone, and posttreatment Boldenone with GSPE). A significant increase in relative body weight, relative heart weight, and hemodynamic parameters, as well as serum concentrations of lactate dehydrogenase, creatine kinase, creatine kinase-muscle brain, myoglobin, cholesterol, low-density lipoprotein cholesterol, risk factor 1/2, K+, and Cl−, in treated rats with Boldenone when compared with control. We also noted a significant increase in the levels of cardiac malondialdehyde, H2O2 generation in heart tissues, mRNA expression of NOX2 and NOX4, and immunoreactivity to proliferating cell nuclear antigen (PCNA). Posttreated rats with Boldenone and GSPE ameliorated cardiac toxicity via inhibition of NOX and a reduction in alteration of the expression of NOX2, NOX4, and PCNA induced by Boldenone. These novel insights into the antioxidative activity of GSPE should serve as a basis for the development of improved chemopreventive or therapeutic strategies for cardiac toxicity.

1. Introduction
The performance-enhancing agents are commonly abused by skilled athletes. Among these agents, many have no proven merits and are linked to serious adverse effects . Androgenic anabolic steroids (AASs), such as Boldenone, are abused to enhance muscle mass, strength, and growth as well as to enhance athletic performance. Many countries have forbidden the use of AAS due to their adverse effects.

The indiscriminate use of Boldenone for enhanced physical performance and muscular appearance in young people is associated with several harmful side effects. Therefore, Boldenone has been classified as “class 2A” (growth promoter and steroid; probable human carcinogen with a high carcinogenic index) by the International Agency for Research on Cancer [3]. The anabolic steroids adverse effects in men include enlarged breast, inhibition of endogenous testosterone, decreased production of sperms, and atrophy of testes [4, 5]. However, a relatively small number of studies have investigated the effects of anabolic steroids on the circulatory system. Cardiovascular diseases are the leading cause of disability and death worldwide and impose a huge burden on affected individuals and society. In young athletes abusing anabolic steroids, acute myocardial infarction may occur without any past history of heart disease.

Heart pathophysiology is characterized by the alteration in the redox signaling xanthine oxidase, cytochrome P-450, or the mitochondrial electron transport chain as a byproduct, or directly by the NADPH oxidase (NOX) family of enzymes plays a role in the generation of reactive oxygen species (ROS) [6].

NOX1–NOX5 and DUOX1/2 (NOX family) are expressed differentially between tissues. These enzymes participate in many cellular procedures including the proliferation of cells, the release of calcium, and biosynthesis of hormone; however, their overexpression is linked to the pathophysiology of several diseases [7]. Further, the role of NOXs as generators of ROS is noteworthy as these are the only enzymes in which ROS generation is the primary and only known function. An increasing amount of data has demonstrated clearly that the expression and activity of NOXs correlate with the development and progression of cardiovascular diseases [8]. Antioxidant systems react with intracellular ROS to produce less reactive compounds. Glutathione peroxidase (GPx) and catalase are indulged in hydrogen peroxide detoxification to produce water or in a glutathione- (GSH-) dependent reaction. Superoxide dismutase (SOD) catalyzes the transformation of superoxide to hydrogen peroxide [9].

Recently, several polyphenolic antioxidants derived from grape seeds have been implicated in protection of cell [10]. Extract of grape seed proanthocyanidin (GSPE) is a rich source of proanthocyanidins. The latter are natural antioxidants composed of various polyphenolic compounds with protective effects against ROS-mediated myocardial ischemia-reperfusion injury and apoptosis [11]. Therefore, the biological activities of proanthocyanidins (antioxidant, anti-inflammatory, and anticarcinogenic) and their protective effects (reduction of mitochondrial damage and apoptosis inhibition) [12] have garnered considerable interest.

The important goals of this work were (i) to demonstrate the involvement of NOX2 and NOX4 in oxidative stress in response to Boldenone administration; (ii) to elucidate the role of NOX2 and NOX4 in mediating pathologic hypertrophy in response to Boldenone administration; and (iii) to establish that GSPE exerts ameliorative effects on the endogenous NOX2 and NOX4 expression in the heart, with roles in the regulation of the redox system.

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