The use of AAS showed a dramatic increase in the recent years especially by young adults aiming to increase their power, their body mass and weight and to have a better appearance improving their self-esteem. Also, AAS have been used for decades by athletes and body builders for the same purpose. On the other hand, many side effects were reported with their illicit abuse (Tasgin et al., 2011).
Our work was performed to assess the toxic effects of AAS on the heart and the skeletal muscles and assessment of the ameliorative effects of silymarin and fenugreek seeds extract.
There was a significant difference between the six groups regarding serum troponin levels. Highest level of troponin was seen in group II (5.5 ± 0.5) while the lowest level was in group IV (0.5 ± 0.1).
The AS treated rats showed a significant elevation in troponin levels. Also, there were non-significant decrease in Silymarin and fenugreek seeds extract. When, silymarin and fenugreek seeds extract were given with AS the levels of serum troponin decreased with no significant differences.
Similarly, Kulaksiz and Sefa (2019) reported an increase in the serum levels of troponin in testosterone treated rats.
Cardiac troponin I (cTnI) is a cardiac enzyme which is released when ether is a damage in the myocytes and considered a sensitive factor of these damages. The levels of this enzyme showed no significant changes in research done on mice treated with silymarin alone (Taghiabadi et al., 2012).
Silymarin could keep the membrane integrity, limiting the leakage of enzymes revealing the cardio-protective effect (Alabdan, 2015)
The activity of cardiac enzyme biomarkers including troponin showed a significant decrease with fenugreek seeds by 27% indicating the cardio-protective effect of fenugreek seeds (Pradeep and Srinivasan, 2018).
Kamble and Bodhankar (2014) explained that the cardio-protective effect of fenugreek seed could be due to trigonelline, saponins, 4 hydroxyisoleucine presence, and high fiber contents.
Also, may be due to fenugreek scavenging properties of the free radical. These properties are attributed to the active hydrogen-donating ability of the hydroxyl substitutions (Murugesan et al., 2011).
As regards to the lipid profile, rats received AS showed a significant elevation in serum cholesterol, triglycerides and LDL levels and a significant decrease in serum HDL levels. Silymarin treated rats showed a significant increase in serum cholesterol and HDL levels. Also, there was elevation in serum triglycerides and LDL levels with no significant differences. In rate treated with fenugreek seeds extract, there was a significant decrease in the serum triglycerides and LDL levels. Also, the serum cholesterol levels decreased but with no significant differences. On the other hand, serum HDL levels increased with significant difference.
Similar to the results of this study was that reported by Silva et al. (2018) who demonstrated that AS could significantly raise the serum triglycerides, cholesterol and LDL levels. Unlike, their effect on HDL that showed a significant reduction in serum level.
The abuse of AS in supra-physiological doses is strongly associated with abnormal levels of plasma lipoproteins showing a decreased level of HDL and increased levels of LDL and cholesterol levels (Albano et al., 2021).
Treatment with fenugreek seeds extract showed improvement in lipid profiles, including decrease in serum triglycerides, total cholesterol, and LDL levels. Also, there was a significant elevation in HDL levels (Al-Chalabi et al., 2019).
This effect could be due to the effect of sapogenins agent that play a role in decreasing the synthesis of cholesterol and fatty acids. Also, they increase the excretion of cholesterol in bile lowering the serum cholesterol levels (Hannan et al., 2003).
Also, Roberts (2011) demonstrated that saponins could decrease the cholesterol levels by inhibiting the absorption of cholesterol and enhance its excretion in bile by forming large particles with bile salts. The excess soluble fibers in fenugreek extract may lead to delay in the absorption of fat and carbohydrate adding to the hypolipidemic effect. Also, the mannose presents in the extract reduce the synthesis of cholesterol.
Metwally et al. (2009) regarding the effect of silymarin on lipid profile, they reported increase in serum cholesterol, LDL, triglycerides and HDL levels.
Gobalakrishnan et al. (2016) explained the different mechanisms for silymarin induced hypo-cholesterolemic effect. Among these effects are the increased the production of LDL receptors on hepatic cells increasing the clearance of plasma LDL. Also, silymarin increases the conversion of cholesterol into bile acid. However, silymarin has no effect on the absorption of cholesterol.
By studying the anti-oxidant effects of AS on the heart and skeletal muscles, this study showed that, the tissue levels of SOD, Catalase and reduced glutathione decreased in AS treated rats compared to the control group. On the other hand, the tissue MDA levels were elevated.
The antioxidant effects of silymarin fenugreek seeds extract were evaluated on the heart, skeletal muscles and showed an increase in tissue levels of SOD, Catalase and reduced glutathione with a decrease in MDA levels.
Both silymarin and fenugreek seeds extract improved the levels of SOD, catalase, reduced glutathione and MDA in AS treated rats.
Similarly, previous studies (Krępa et al., 2011 and Tothova et al., 2013) reported that an increase of the lipid peroxidation and decrease of the enzymatic activity of glutathione reductase and SOD of different tissues in adult male rats because of exogenous testosterone intake.
Frankenfeld et al. (2014) showed a decrease in the activity of catalase enzyme after AS (nandrolone decanoate) intake.
High doses of nandrolone decanoate are metabolized by cytochrome P450 mono-oxygenases. This results in production of reactive oxygen species leading to the upregulation followed by exhaustion of the antioxidants enzymes activity (Pey et al., 2003).
Furthermore, anabolic steroids enhance the activity of lipase enzyme which in turn increases the rate of lipolysis (Langfort et al., 2010). which in turn increases the availability of long chain fatty acids for mitochondrial oxidation and production of ATP leading to development of the lipid peroxidation and generation of ROS (Niki et al., 2005).
As regards to silymarin, our results was in consistence with the study performed by Avci et al. (2017) about the effect of silymarin on MDA and SOD levels in the heart of rats. They demonstrated an increase in the activity of SOD and decrease in MDA levels denoting a protective effect against lipid peroxidation and oxidative damage.
Similarly, Aktas and Ozgocmen (2020) revealed in their study an increase in the activity of glutathione enzyme and decrease in MDA levels in cardiac tissue of rats treated by silymarin.
Surai (2015) reported that silymarin is a potent antioxidant inhibiting the lipid peroxidation and prevents the reduction of glutathione enhancing the activity of antioxidants enzymes.
This effect could be clarified by its effect on maintaining the integrity of cell membranes against the oxidative damage of ROS (Nencini et al., 2007).
This antioxidant properties probably due to the presence of silydianin, silybin, silychristin and flavolignans which are chemical bioactive compounds present in silymarin. (Yaman et al., 2018).
Regarding fenugreek seeds extract, Bafadam et al. (2021) revealed an increase in the activity of SOD and catalase in heart tissue of rats and decrease in lipid peroxidation augmented by the decreased MDA levels in hearts tissues of rats treated with germinated fenugreek seed. Also, Arshadi et al. (2015) showed an increase in the activity of cardiac antioxidant enzymes like reduced glutathione, catalase and SOD.
Fenugreek seeds extract antioxidant properties are mainly because of the presence of the biologically active compounds flavonoids and polyphenols. Its major active antioxidant compounds are isovitexin and flavones of vitexin (Khole et al., 2014).
Histopathological examination of cardiac muscle in our study showed that, the most commonly observed lesions were degenerative changes and cardiac muscles necrosis (hyalinosis) which was severe in rats treated with AS. On the other hand, normal histological structure of cardiac muscles could be found in rats of the control group and those received silymarin and fenugreek seeds extract. Improvement of the toxic effects induced by AS was observed with treatment of silymarin and fenugreek seeds extract with mild degenerative changes were seen with treatment by fenugreek seeds extract and moderate degenerative changes with treatment by silymarin.
Histopathological examination of the skeletal muscle in control group rats and rats treated with silymarin and fenugreek seeds extract were normal. Severe degenerative changes associated with nuclear pyknosis of the sarcoplasm were seen in rats treated with nandrolone decanoate. The toxic effect of AS could be improved with treatment by silymarin and fenugreek seeds extract.
Also, Kahal and Allem (2018) showed in their study elongation, severe degeneration and may be rupture of the cardiac muscle. Also, these results coincide with Hassan et al. (2009) “ AS sustanon induces severe ischemic necrosis and degeneration of the cardiac muscle fibers in male albino rats.”
Abdelhafez (2014) showed highly degenerated muscle fibers with areas of hemorrhage and widened endomysium. Also, she demonstrated a numerous pyknotic and karyolytic nuclei.
Also, Elgendy et al. (2018) reported hypertrophy and degeneration of both cardiac and skeletal muscles and explained this by its effect on the androgen receptors that are widely distributed in different types of muscles.
The defensive effect of silymarin against cardiac damage was studied by Avci et al. (2017) who reported that silymarin improve the cardiotoxic histopathological effects induced by cyclophosphamide.
Other study performed by Aktas and Ozgocmen (2020) showed that silymarin preserves the histological structure of the heart and effectively promotes the antioxidant defense system against valproic acid induced injuries.
Also, El-Shitany et al. (2008) demonstrated an improvement in the histopathological effects in Adriamycin induced cardiotoxicity when combined with silymarin and explained this by the antioxidant activity of silymarin inhibiting the lipid peroxidation and promoting antioxidant enzymes activity.
Also, Mendoza et al. (2020) demonstrated a protective effect of silymarin on cardiac and skeletal muscle injuries due to its potent antioxidant effects.