BENHA VETERINARY MEDICAL JOURNAL, VOL. 35, NO. 2: 237-249, DECEMBER , 2018

BENHA UNIVERSITY FACULTY OF VETERINARY

Ameliorative Effects of Lycopene against 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 like boldenone (BOL) may increase lean muscle mass, strength, and the ability to train longer and harder, the serious side effects of 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 used as anabolic steroids in livestock hormone) that derived from , 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, 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 which therapy is , 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 (Tousson 2.1. Experimental animals and dosing et al; 2012a). Injection of the 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 (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, globulin and albumin/globulin compared to control and lycopene groups, but (A/G) ratio, non-significance change in was in co-treated group (Lycopene + Boldenone) detected in Lycopene group compared to the amelioration effect of lycopene is more control group. While, there was significant clear (Table 3). Regarding the results of LDH and CK increase in total protein and globulin and significant decrease in albumin and A/G ratio showed non-significant changes in lycopene 732

Sabra et al. (2018). BVMJ-35(2): 237-249 group compared to control group, while the experiment, this changes as follows:- In heart, results of boldenone group compared to sever dilatation in myocardial blood control and lycopene groups showed capillaries associated with focal haemorrages significant increase in LDH and CK. But, the in between myocardial bundles photo 2 (c, d). results of co- treated group (Lycopene + In skeletal muscles showed focal Boldenone) were ameliorated when compared hyalinization some of the bundles associated to others groups (Table 4). with corrugation in the outside of some There was no significant change in others. Photo 3 (c, d). In the liver focal total protein of skeletal muscles in Lycopene necrosis was detected in the hepatic group compared to control group. While, parenchyma associated with dilatation in the there was significant increase in total protein central vein as well as fatty change in the of skeletal muscles in Boldenone group surrounding adjacent hepatocytes , the portal compared to control and Lycopene groups. area showed inflammatory cells infiltration But, the results of treated group (Lycopene + and few fibroblastic cells proliferation photo Boldenone) were ameliorated when compared 1 (c, d, e and f). On other hand, there is a to others groups (Table 5). markedly ameliorative effect in the group Histopathological examination of received (Lycopene + Boldenone) compared liver, heart and skeletal muscles of rats in with Boldenone group as the control, Lycopene and (Lycopene + histopathological photo 1 (g), photo 2 (e) and Boldenone) groups, showed the normal photo 3 (e) showed no histopathological histopathological structure of examined alternations in liver and skeletal muscles organs and no histopathological alterations tissues, but in heart showed only a mild after 5 weeks of the experiment photos (1-3). dilatation in the myocardial blood capillaries. But in BOL group rats showed histopathological changes after 5 weeks of the

Table (1): Effect of Lycopene and/or Boldenone Undecylenate on body weight (gm) (Mean ± SE) (N=5). Before After treatment (week)(gm) Group treatment 1st 2nd 3rd 4th Control 127.8±2.91a 121.4±1.94c 119.2±2.92d 117.0±4.05c 115.4±2.77d Lycopene(LYCO) 130.8±2.75a 135.4±2.96b 145.6±3.56c 159.4±8.43b 173.2±15.40c Boldenone(BOL) 130.8±2.52a 162.4±7.58a 182.6±6.11a 184.4±6.81a 195.4±6.93a

BOL+LYCO 128.6±0.93a 160.8±2.71a 171.6±3.85b 179.6±4.23a 187.4±5.41b Mean with different superscript letters at the same row are significantly different at (P<0.05).

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Ameliorative Effects of Lycopene against Boldenone Undecylenate Toxicity in Albino Rats

Table (2): Effect of Lycopene and/or Boldenone Undecylenate on total protein, albumin, globulin and A/G ratio (mean±SE) (N=5). Total Group Albumin(g/dl) Globulin(g/dl) A/G ratio(%) protein(g/dl) Control 4.95±0.07c 4.03±0.22a 0.91±0.20b 4.42±1.49a Lycopene(LYCO) 5.69±0.13bc 3.78±0.15a 1.91±0.25b 1.98±0.50b Boldenone(BOL) 7.16±0.20a 3.13±0.41a 4.03±0.38a 0.77±0.24b BOL+LYCO 6.66±0.76ab 3.61±0.32a 3.05±0.50a 1.18±0.15b Mean with different superscript letters at the same row are significantly different at (P<0.05).

Table (3): Effect of Lycopene and/or Boldenone Undecylenate on MDA and SOD (mean±SE) (N=5). Group MDA(nmol/ml) SOD(u/ml) Control 1.95±0.22b 156.25±22.45a Lycopene(LYCO) 2.62±0.60ab 178.40±25.53a Boldenone(BOL) 3.34±0.20a 73.63±10.29b BOL+LYCO 2.46±0.10ab 101.00±4.72b Mean with different superscript letters at the same row are significantly different at (P<0.05).

Table (4): Effect of Lycopene and/or Boldenone Undecylenate on LDH and CK (mean±SE) (N=5). Group LDH(u/l) CK(u/l) Control 837.40±82.50b 449.60±131.11b Lycopene(LYCO) 826.00±73.00b 356.00±62.86b Boldenone(BOL) 1055.80±7.00a 905.80±12.78a BOL+LYCO 919.20±69.18ab 756.80±123.20a Mean with different superscript letters at the same row are significantly different at (P<0.05).

Table (5): Effect of Lycopene and/or Boldenone Undecylenate on total protein in tissues (skeletal muscles) (mean±SE) (N=5). Group Total protein(g/dl) Control 1.20±0.06b Lycopene(LYCO) 1.32±0.29b Boldenone(BOL) 2.53±0.20a BOL+LYCO 1.53±0.10b Mean with different superscript letters at the same row are significantly different at (P<0.05).

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Sabra et al. (2018). BVMJ-35(2): 237-249 Photo (1) Liver

photo (a): Liver of rat in control group Photo (b): Liver of rat in lycopene group showing the normal histological structure of showing the normal histological structure the central vein and surrounding hepatocytes H&E (X 40). in the hepatic parenchyma H&E (X 40).

Photo (c): Liver of rat in boldenone group Photo (d): Liver of rat in boldenone group showing dilatation in central vein H&E (X showing focal necrosis in hepatic 40). parenchyma H&E (X 40).

Photo (e): Liver of rat in boldenone group Photo (f): Liver of rat in boldenone group showing mild fatty change in hepatocytes showing inflammatory cells infiltrations in surrounding the dilated central vein H&E (X the portal area H&E (X 80). 80).

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Ameliorative Effects of Lycopene against Boldenone Undecylenate Toxicity in Albino Rats

Photo (g): Liver of rat in lycopene plus boldenone group showing the normal histological structure H&E (X 40).

Photo (2) Heart

Photo (a): Heart of rat in control group Photo (b): Heart of rat in lycopene group showing normal histological showing normal histological structure of structure of myocardial bundles myocardium H&E (X 40). H&E (X 40).

Photo (c): Hearts of rat in boldenone group Photo (d): Heart of rat in boldenone group showing sever dilatation in cardiac blood showing hemorrhage in between myocardial vessels in myocardium H&E (X 40). bundles H&E (X 40).

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Sabra et al. (2018). BVMJ-35(2): 237-249

Photo (e): Heart of rat in lycopene plus boldenone group showing mild dilatation in myocardial blood capillaries H&E (X 40).

Photo (3) Skeletal muscle

Photo (a): Skeletal muscle of rat in control Photo (b): Skeletal muscle of rat in group showing normal histological structure lycopene group showing normal as normal striated muscle bundles H&E (X 40). histological structure H&E (X 40).

Photo (c): Skeletal muscle of boldenone Photo (d): Skeletal muscle of boldenone group showing focal hyalinization of some group showing corrugations in outer line skeletal muscle bundles H&E (X 40). of muscle bundles H&E (X 40).

Photo (e): Skeletal muscle of rat in lycopene plus boldenone group showing normal histological structure H&E (X 40). 744

Ameliorative Effects of Lycopene against Boldenone Undecylenate Toxicity in Albino Rats

4. DISCUSSION

Boldenone undecylenate one of AAS administration. These findings could be used to improvement of the growth and food attributed to the nitrogen-retention capacity of conversion in food producing animals. The the drug. Additionally, the increment of anabolic effects of AAS promote protein globulins may be attributed to the increased synthesis, muscle growth and erythropoiesis α- and β-fractions rather than the γ-fractions, (Soma, et al., 2007). Hence, AAS are used to including positive acute-phase proteins. enhance strength and durability of canine, Albumin, in contrast, is considered a negative equine and human athletes (Matinhomae, et acute-phase protein that is decreased in al., 2014). Our study aimed to investigate the inflammation, especially hepatic. Although role of lycopene as antioxidant in improving the calculated albumin/globulin ratio was the oxidative stress induced by boldenone in decreased, albumin concentrations showed male albino rats. insignificant changes in our study and this Our results revealed that injection of result was in agreement with (Abdelrazzag, et the anabolic steroid BOL to albino male rats al., 2016). Total protein significantly evoked a significant increase in growth rate as increased with the BOL injection due as indicated by a significant increase in total mentioned previously to the multifactorial weight gain (TWG), similar findings were effects of BOL on the protein metabolism reported by (Saleh and Waded, 2014) and (ElSayed and Kerdasy, 2007). Our data This effect could be attributed to the fact that confirmed by the histopthology of liver of BOL act upon the androgen receptors in BOL group showed that rats that received five anabolic-responsive tissues promoting the doses of boldenone revealed focal necrosis body tissue building processes due to was detected in the hepatic parenchyma increased protein synthesis. This is achieved associated with dilatation in the central vein by converting the negative nitrogen balance as well as fatty change in the surrounding into a positive one through improving the use adjacent hepatocytes, the portal area showed of ingested proteins and increasing nitrogen inflammatory cells infiltration and few retention and animal’s appetite (Falanga et al., fibroblastic cells proliferation in liver photo1 1998). Others reported that anabolic (c, d, e, f). hormones can increase the cellular protein Elevated levels of CK and LDH in biosynthesis indirectly via stimulation of this study indicated an adverse effect of growth hormone and insulin like growth boldenone on cardiac and/or skeletal muscle. factor secretion (Arnold et al., 1996). This confirmed by histopathological Furthermore, it was also assumed that AAS abnormalities in heart and skeketal muscles in reduce recovery time by blocking the effects BOL group. In heart, sever dilatation in of stress hormone cortisol on muscle tissue, myocardial blood capillaries associated with so that catabolism of muscle is greatly focal haemorrages in between myocardial reduced (Sauerwein et al; 1991). bundles photo2 (c- d). In skeletal muscles Although hypo- proteinemia is an showed focal hyalinization some of the established finding in liver damage (Larrey, bundles associated with corrugation in the 2002), given that the liver creates most outside of some others photo3 (c- d). Multiple plasma-protein fractions, here, the total investigations reported that boldenone abuse plasma protein and globulin concentrations was associated with occurrences of serious were significantly increased after boldenone cardiovascular events in young athletes,

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Sabra et al. (2018). BVMJ-35(2): 237-249 including development of cardiomyopathy, inhibitory effect of boldenone on enzymes atrial fibrillation, infarction, disturbances of activities. Our results are in agreement with hemostatic system, and ventricular (Mayada, et al., 2015) who reported that thrombosis (Hartgens and Kuipers, 2004). On Intramuscular injection of BOL increased other hand, there is a markedly ameliorative malondialdehyde (MDA) level, but markedly effect in the group received lycopene plus lowered activities of superoxide dismutase BOL compared with BOL group as the (SOD) and the findings of (El-Moghazy et al. histopathological pictures showed no 2012) who reported that BOL injection in histopathological alternation in the skeletal rabbits caused hepatic oxidative stress. These muscles tissue photo3 (e), and showed a mild findings are also concordant with previous dilatation in the myocardial blood capillaries studies by (Sadowska-Krepa et al. 2011), photo2 (e). where they cited that AAS treatment induced Oxidative stress has been defined as a oxidative stress in rats. Our data confirmed disturbance in the balance between the histopthology of liver of BOL group antioxidants and pro-oxidants (free radicals showed that rats that received five doses of and other reactive species) with increased boldenone revealed focal necrosis was levels of pro-oxidants leading to potential detected in the hepatic parenchyma associated damage. This imbalance can be an effect of with dilatation in the central vein as well as depletion of endogenous antioxidants, low fatty change in the surrounding adjacent dietary intake of antioxidants and/or increased hepatocytes, the portal area showed formation of free radicals and other reactive inflammatory cells infiltration and few species. In human tissues, a condition of fibroblastic cells proliferation in liver photo1 oxidative stress can be revealed through (c, d, e,f). Interestingly, the ameliorative searching for specific biomarkers of oxidative effect of lycopene against effect of BOL on damage to lipids, proteins and nucleic acids MDA and SOD is clearly observed. MDA (Zhang, et al., 2009). BOL treatment can level was reduced and SOD was elevated, this cause an oxidative stress situation in liver as was in comparison between co- treated group indicated by enhanced MDA activity (LYCO plus BOL) and BOL group, this (increased MDA concentration). Interestingly, ameliorative effect in the group received reactive oxygen species (ROS) and MDA lycopene plus BOL confirmed by the have been implicated in the pathogenesis of histopathological Photo1 (g) showed no many types of liver injury and especially in histopathological alternation in liver. the hepatic damage induced by several toxic In our study, total protein in skeletal drugs (Uchida, et al; 1999). Our results are in muscles was significantly increased, this was agreement with (Lamiaa et al; 2015) who attributed to the mechanism of action of reported that a significant increase in the boldenone steroid hormones work by MDA, NO, GSH, total protein, total thiol and stimulation of receptors molecule in muscle catalase and significant decreased in co- cells, which activate specific genes to produce treated boldenone with propolis in rat liver proteins. They also affect the activation rate and kidney when compared to the boldenone of enzyme system involved in protein group. In the current study; decrease in the mechanism. Thus enhancing protein synthesis activities of antioxidant enzymes SOD can be and inhibiting protein degradation (Mottram explained either with their induction during and George, 2000). Also BOL increases the conversion of free radicals into inactive muscle size owing to promotion of positive metabolites or secondarily with the direct nitrogen balance by stimulating protein

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Ameliorative Effects of Lycopene against Boldenone Undecylenate Toxicity in Albino Rats production and reducing protein destruction, of testosterone. J Endocrinol.; as well as causing retention of body water, 150(3):391-9. nitrogen, sodium, and potassium and calcium Banchroft, J.D. Stevens, A. And Turner, ions (Tousson, et al., 2012b). Our result D.R. (1996): Theory And Practice Of confirmed by histopathological abnormalities Histological Techniques. Fourth Ed. in skeketal muscles photo 3 (c,d) in BOL Churchil Livingstone, New York, group showed focal hyalinization some of the London, San Francisco, Tokyo. bundles associated with corrugation in the Ben-Dor A, Nahum A, Danilenko M, Giat Y, outside of some others. But there is a Stahl W, Martin HD, Emmerich T, Noy markedly ameliorative effect in the group N, Levy J, Sharoni Y; (2001): Effects of received lycopene plus BOL compared with acyclo-retinoic acid and lycopene on BOL group as the histopathological pictures activation of the retinoic acid receptor showed no histopathological alternation in and proliferation of mammary cancer skeketal muscles photo3 (e). Our cells. Arch Biochem Biophys.; histopathology study was in agreement with 391(1):295–302. (Tousson, 2013) who reported that rabbits Britton G. (1995): Structure and properties of that received BOL showed hypertrophy, carotenoids in relation to function. nuclear pycnosis and cytoplasmic vacuoles in FASEB J; 9: 1551-8. both skeletal and cardiac muscles, Broody, S. (1945): Bioenergetis and Growth disturbances of the hepatocytes radially Reinhold pub Crop. New york. arranged cords with multifocal hepatocellular Burtis CA, Ashwood ER, Bruns DE, WB vacuolations in the liver. Saunders Co, (2005): Tietz Textbook of Clinical Chemistry and Molecular 5. CONCLUSION Diagnosis, 4th ed. This study concluded that the Cannizzo, T.F., Zancanaro, G., Spada, F., Boldenone is one of steroidal substance Mulasso, C. and Biolatti, B. (2007): induce increase of body weight, changes in Pathology of the testicle and sex haemopiotic system and hepatotoxicity also, it accessory glands following the concluded that lycopene has protective effect administration of boldenone and against Boldenone toxicity, as it showed as growth promoters in veal marked improvement in haematological calves. J. Vet. Med. Sci., 69: 1109- alterations and oxidative changes in liver, 1116. heart and skeletal muscle. Chitra, KC. Latchoumycandane, C. and Mathur, pp. (1999): chronic-effect of 6. REFERENCES Abdelrazzag Elmajdoub, Aboubaker Garbaj, on the testicular functions Said Abolghait, Abubakr El- of rat. Asian J Androle; 1:203-6. Mahmoudy, (2016): Evaluation of Davis, B. (1964): Disk electrophoresis. II boldenone as a growth promoter in Method and application to human broilers: safety and meat quality serum protein. Annual of New York aspects, science direct 24 284 e 292. Academy Science, 929: 404-427. Arnold AM, Peralta JM, Thonney ML. De Brabander, H.F., Poelmans, S., Schilt, R., (1996): Ontogeny of growth hormone, Stephany, R.W., Le Bizec, B.,Draisci, insulin-like growth factor-I, R., Sterk, S.S., van Ginkel, L.A., Courtheyn, D., Van Hoof, N., Macrì, and cortisol in the growing lamb: effect A.and De Wasch,K. (2004): Presence

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Ameliorative Effects of Lycopene against Boldenone Undecylenate Toxicity in Albino Rats

Testosterone Propionate Treatment testis of rabbits after injection with the Reverses the Effects of Endurance growth promoter boldenone. Reprod. Training on Myocardial Antioxidant Sci., 19: 253-259. Defenses in Adolescent Male Rats. Tousson, E., El-Moghazy, M., Massoud, A. Cardiovasc. Toxicol, 11: 118-127. and Sakeran, M. (2012b): Changes in Saleh N. and Waded E. (2014): Immune hepatic and renal structure and function response following the administration after a growth promoter boldenone of the anabolic steroid Boldenone injection in rabbits. 62: 171-180. Undecylenate in rabbits. Stem Cell; Uchida, M., Shiraishi, K., Naito, Y., Torii, 5:80 e 7. Y., Nakamura, Y. and Osawa, T. Sauerwein H, Diirsch I, Meyer H. (1991): (1999): Activation of stress signaling Quantitation of glucocorticoid receptors pathways by the end product of lipid in bovine skeletal muscle: topographical peroxidation. 4-Hydroxy-2-nonenal is a distribution, sex effect and breed potential inducer of intracellular comparisons. J Steroid Biochem.; peroxide production, J. Biol. Chem. 39:941-45. 274, 2234– 2242. Shahidi NT. (2001): A review of the Zedan NS, Tousson E, Massoud A and El- chemistry, biological action and clinical Saeed A. (2014): Biochemical and applications of anabolic-androgenic histopathological alterations in rattestes steroids. Clin. Therapeutics. 23:1355– after injection with the growth promoter 1390. Equigan with reference to the Soma, L.R., C.E. Uboh, F. Guan, S. MC- ameliorating role of Proplis. 2nd Donnell and J. Pack, (2007): International Conference for Nutrition of boldenone and and Growth, Barcelona, Spain. stanozolol and the results of Zhang, X.L., Yang, Y.S., Ping, D., Hui, J., quantification of anabolic and Zhou, M Gong, Y. and Huang, J. androgenic steroids in race horses and (2009): Comparative Study on non-race horses. J. of Veterinary Overexpression of HER2/neu and Pharmacology and Therapeutics. 30: HER3 in Gastric Cancer. World Journal 101-108. of Surgery. 33: 2112-2118. Sönmex M, Turk G, Cerbasi AO, Sakin F, Atessahin A. (2011): Attenuating effect Of lycopene And ellagic Acid On 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin- Induced spermiotoxicity And testicular apoptosis. Drug Chem Toxicol; 34: 347-356. Tousson E. (2013): Histopathological alterations after a growth promoter boldenone injection in rabbits. Toxicology and Health; 32:299 e 305. Tousson, E., El-Moghazy, M., Massoud, A. and Akela M., (2012a): Histopathological and immunohistochemical changes in the

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