BENHA VETERINARY MEDICAL JOURNAL, VOL. 35, NO. 2: 237-249, DECEMBER , 2018 BENHA UNIVERSITY FACULTY OF VETERINARY MEDICINE Ameliorative Effects of Lycopene against Boldenone Undecylenate Toxicity in Albino Rats Sabra H.M., El-Shawarby R.M., Abosalem M., Nabila M. A., Ibrahim S.S. Department of Forensic Medicine & Toxicology. Faculty of Veterinary. Medicine. Benha University. A B S T R A C T Although anabolic androgenic steroids like boldenone (BOL) may increase lean muscle mass, strength, and the ability to train longer and harder, the serious side effects of steroid are many and may not be reversible. So, the present study was designed to investigate the role of lycopene in improving the oxidative stress of boldenone in male albino rats. Twenty eight male adult rats were equally divided into four groups.1st group (control) that received olive oil once a week (5 mg/kg intramuscularly). 2nd group includes rats received lycopene daily (orally, 60mg/kg body weight). 3rd group includes animals that received intramuscular injections of boldenone once a week (5 mg/Kg body weight). 4th group was co-treated with boldenone and lycopene with the same previous doses of both, the experiment extended for 5 weeks. Intramuscular injection of rats with caused significant increase in total weight gain (TWG), serum total protein and globulin, malandialdehyde (MDA), Lactate dehydrogenase (LDH), creatinine kinase (CK), total protein in skeletal muscle, when compared with the control group. On the other hand, a significant decrease in albumin, Albumin/Globulin (A\G) ratio and superoxide dismutase (SOD) in boldenone group when compared with the control group. Also, the result of rats administered lycopene plus Boldenone (group4) showed marked improvement in all tested parameters, these findings may be attributed to that Lycopene has an effect on regulation of intracellular gap- junction communication, genes function regulation, modulation of various metabolic pathways of xenobiotic, hormone and immune modulation. These findings suggested that misuse of boldenone may contribute to a continuous tissue damage, so young people -especially- should be careful if they want to use boldenone to enhance their strength and endurance or if they want to use it in the farm animal as growth promoter. This showed that the desired dose of boldenone can safely be used with lycopene in improving the oxidative stress of boldenone. Key words: Boldenone toxicity, Lycopene, Oxidative stress, Antioxidants, Liver, Heart. (http://www.bvmj.bu.edu.eg) (BVMJ-35(2): 237-249, 2018) 1. INTRODUCTION Boldenone (1, 4-androstadiene-17beta-ol- (Cannizzo et al; 2007). BOL is one of 3-one; BOL) and its precursor boldenone are anabolic steroids (synthetic androgen used as anabolic steroids in livestock hormone) that derived from testosterone, 732 Sabra et al. (2018). BVMJ-35(2): 237-249 differs from testosterone by only one double lycopene is transported in the blood by bond at the 1-position (Zedan et al; 2014). various lipoproteins and accumulates in the BOL has a very long half-life and can show liver, adrenal glands, prostate and testes up on a steroid test for up to 1.5 years. BOL (Nahum et al; 2001). Of all the carotenoids, has been detected in untreated animals of lycopene is one of the most potent several animal species (De Brabender et al antioxidants; particularly able to quench 2004). BOL has been developed for singlet oxygen and peroxyl radicals, both of veterinary use. BOL improves growth and which are thought to be responsible for feed conversion in veal calves and therefore damaging DNA in a process which can lead might be used illegally to achieve more to the initiation of cancer. Because of this efficient meat production (Soma et al; 2007). ability, lycopene is being investigated as a Boldenone has dual effects on humans, both potential agent for the prevention of some directly and indirectly; directly as injection to types of cancers, particularly prostate cancer build muscles and indirectly as through (Di Mascio, et al; 1989). Lycopene protects consuming meat of animals that where treated against oxidative damage to lipids, proteins, with boldenone. In addition to the growth and DNA. Lycopene is a potent quencher of promoting effects, anabolic steroids have singlet oxygen (a reactive form of oxygen), been shown to adversely affect the which suggests that it may have cardiovascular, hepatic, kidney and endocrine comparatively stronger antioxidant properties systems (Tousson et al; 2012b). A major side than other major plasma carotenoids. effect of anabolic- androgenic steroids Lycopene induces Phase II enzymes which therapy is hepatotoxicity, including elevated help to eliminate carcinogens and toxins from levels of liver enzymes, cholestatic jaundice, the body (Ben-Dor et al 2001). peliosishepatis and various neoplastic lesions Aim of work: (Shahidi 2001). Boldenone increases muscle Our study aimed to investigate the role of size due to promotion of positive nitrogen lycopene as antioxidant in improving the balance by stimulating protein production and oxidative stress induced by boldenone in male reducing protein destruction, moreover it albino rats. produces retention of body water, nitrogen, 2. MATERIALS AND METHODS sodium, potassium and calcium ions (Tousson 2.1. Experimental animals and dosing et al; 2012a). Injection of the anabolic steroid Twenty eight apparently healthy male boldenone induced changes in oxidative stress Wister albino rats weighed 125-160 gm biomarker levels and antioxidant defense obtained from Veterinary National Research systems in rabbit liver and kidney (Mottram Institute, Cairo, Egypt, were randomly and George, 2000). divided into 4 groups each one contains 7 rats. Lycopene is a bright red carotenoid pigment 1st group (control) includes animals that found in tomatoes and other red fruits injected intramuscularly with olive oil once a (Britton, 1995). Lycopene is an important week. 2nd group was rats received lycopene intermediate in the biosynthesis of many daily (orally, 60mg/kg body weight)-obtained carotenoids, including beta-carotene, which is from Al Debeiky pharmaceuitical company- responsible for pigmentation, photosynthesis, according to (Sönmex et al., 2011). 3rd group and photo-protection. Lycopene's eleven includes animals that received intramuscular conjugated double bonds give it its deep red injections of boldenone once a week (5 color and are responsible for its antioxidant mg/Kg body weight) - obtained from Swiss activity. When absorbed from the stomach, labs- according to (Gabr, et al., 2009). 4th 732 Ameliorative Effects of Lycopene against Boldenone Undecylenate Toxicity in Albino Rats group was co-treated with boldenone as (albumin ÷ globulin), Antioxidant enzyme (intramuscular injections, 5 mg/Kg body activity: MDA was determined by the weight) plus lycopene (orally, 60mg/kg body reaction described by ohkawa et al; (1979), weight). The experiment was extended for 5 SOD was performed according to Oyanagui, weeks. (1984). CK and LDH enzymes concentrations 2.2. Performance and growth measurement are determined and measured according to Mean body weight was estimated Burtis, et al; (2005). every week, while the body gain was 2.5. Determination of total protein skeletal calculated according to (Broody; 1945). muscle by electrophoresis: 2.3. Sampling The method used for electrophoresis Serum obtained by blood collection was that of Davis, (1964). after the end of experiment by 24 hours from The electropherogram calculated the inner canth of eye in non-heparinized dry according to SynGene S. No.17292*14518 clean centrifuge tube then centrifugation, the sme*mpcs. serum was kept at -20oC till used in the 2.6. Histopathological examination: evaluation of biochemical studies. Tissue Autopsy samples were taken from the samples (liver, heart and skeletal muscle) liver, heart and skeletal muscle in different were collected after the end of experiment by groups of rats and used for histopathological 24 hours and fixed in 10% formalin for examination using H& E stain according to histopathological studies. part of skeletal Banchroft et al., (1996). muscle kept in -80o for total protein 2.7. Statistical analysis: electrophoresis. Data were statically analyzed by using 2.4. serum Biochemical parameters: the statistical software package SPSS for Total protein, was determined windows (Version 16). The significance of calorimetrically according to Gornall, et al; differences between more than two groups (1949), albumin, was determined was evaluated by one way analysis of calorimetrically according to Doumas, et al., variance (ANOVA) using DUNCAN as post- (1971), globulin is calculated as (total protein hoc test according to spss1nc. Chicago, USA. – albumin= globulin), A/G ratio is calculated in Boldenone group compared to control 3. RESULTS Body weight showed a significant group and Lycopene group. The result of co- increase from 1thweek till 4thweek in the treated group (Lycopene + Boldenone) Boldenone group when compared with showed amelioration in in comparison with control group and Lycopene group. While other groups (Table 2). there was significant decrease in the body The result of MDA and SOD showed weight of treated group which received no significant changes in lycopene group Lycopene plus Boldenone in compared with compared to control group. In Boldenone the boldenone group (Table 1) . group, the results showed significant increase Regarding the result of total protein, in MDA and significant decrease in SOD in albumin,