International Journal of Scientific and Research Publications, Volume 6, Issue 1, January 2016 408 ISSN 2250-3153 Captopril and telmisartan ameliorate cisplatin-induced testicular damage in rats via anti-inflammatory and antioxidant pathways Ahmed H. Eid*, Noha F. Abdelkader**, Ola M. Abd El-Raouf*, Hala M. Fawzy*, Ezz-El-Din S. El-Denshary** *Department of Pharmacology, National Organization for Drug Control and Research (NODCAR), Giza, Egypt **Department of Pharmacology and Toxicology, Faculty of Pharmacy, Cairo University, Cairo, Egypt. Abstract- Despite the prevalent clinical applications of Male reproductive toxicity rises from targeting rapidly cisplatin, serious toxic side effects comprising reproductive growing testicular cell types such as leydig, sertoli, and germ toxicity confine its therapeutic efficacy. Thus, the current study cells (Boekelheide, 2005). Cisplatin adversely impairs non target explored the possible protective effect of captopril and tissues via increased free radical generation and depletion of telmisartan against cisplatin-induced testicular damage in rats. antioxidants; which results in inhibition of protein synthesis and Captopril (100 mg/kg) and telmisartan (10 mg/kg) were orally DNA damage (Fung and Vaughn, 2011, Ilbey et al., 2009a). administered for 15 days, whereas cisplatin (10 mg/kg; i.p.) was Growing evidence verifies that activation of the renin angiotensin injected as a single dose at the 12th day to induce testicular system (RAS) is largely implicated in cisplatin-induced tissue damage in adult male Sprague-Dawley rats. Cisplatin injury (El-Sayed et al., 2008, Saleh et al., 2009) where cisplatin prominently decreased reproductive organs weights, sperm amplifies RAS components level both experimentally (El-Sayed count, sperm motility, and increased sperm abnormalities, along et al., 2008, Okui et al., 2012) and clinically (Kurt et al., 1999). with histopathological damage of testicular tissues. In addition, it Indeed, angiotensin II, the chief component of RAS, induces resulted in a significant decline in serum testosterone as well as tissue oxidative stress and apoptosis (Cai et al., 2003, Wolf, testicular enzymatic and non-enzymatic antioxidants levels 2000). Besides, it triggers inflammation via upregulation of pro- (superoxide dismutase, catalase, glutathione peroxides, and inflammatory transcription factors, which results in a vast array reduced glutathione), parallel to a remarkable elevation in of proinflammatory cytokines that further injures the tissues testicular content of malondialdehyde, nitric oxide, tumor (Hayashi et al., 2010). necrosis factor-α, and nuclear factor-kappa B. Treatment with Therefore, the current study was conducted to investigate the captopril or telmisartan markedly attenuated cisplatin-induced efficacy of RAS blockade against cisplatin-induced testicular injury by suppression of oxidative/nitrosative stress and damage. Captopril is an angiotensin converting enzyme inhibitor inflammation, amendment of antioxidant defenses, as well as that reduces circulating and tissue levels of angiotensin II. improvement of steroidogenesis, spermatogenesis and testicular Moreover, it is a powerful thiol-containing antioxidant (Liu et histological features. This study suggests a novel therapeutic al., 2007) that possesses anti-inflammatory potential application for captopril and telmisartan as protective agents (Amirshahrokhi et al., 2010). Captopril is widely used for the against cisplatin-induced testicular toxicity through their treatment of hypertension, congestive heart failure, and promising anti-inflammatory and antioxidant capacities. diabetic nephropathy (Cohen et al., 1996). In context, telmisartan is a highly selective angiotensin II type-1 receptor Index Terms- Captopril, cisplatin, oxidative stress, blocker which is approved for the treatment of hypertension. It is telmisartan, testicular damage. endowed with potent anti-inflammatory, antioxidant and anti- apoptotic effects (Fouad et al., 2010). I. INTRODUCTION Interestingly, recent studies revealed that both drugs isplatin is an extremely effective anticancer drug for the ameliorated chemotherapy-induced cardiotoxicity and C treatment of diverse types of solid tumors (Amin and nephrotoxicity (Abd-Ellah, 2012, Cadeddu et al., 2010, Buratovich, 2009). Despite the improvement in quality of life of Ibrahim et al., 2009) as well as cadmium-induced testicular cancer patients; the use of cisplatin was restricted clinically by its toxicity (Fouad and Jresat, 2013). Herein, the potential major side effects including testicular toxicity (Beytur et al., protective effects of captopril and telmisartan against testicular 2012, Fung and Vaughn, 2011). In adult men, fertility can be toxicity induced by cisplatin in male rats were investigated by the preserved by spermatozoa cryopreservation and intracytoplasmic assessment of hormonal and spermatological changes, oxidative sperm injection, however, these methods are not feasible options stress parameters, inflammatory markers, and the testicular for pre-pubertal patients (Rezvanfar et al., 2013). Besides, in histopathological alterations. adults, the freezing and thawing of semen can reduce the sperm quality (Maines et al., 1990). Interestingly, growing evidences II. MATERIALS AND METHODS revealed that combination therapy can be beneficial to overcome Animals this special reproductive toxicity (Narayana et al., 2012, Adult male Sprague-Dawley rats; weighing 240-280 g, were Ahmed et al., 2011). obtained from the breeding colony of the National Organization for Drug Control and Research (NODCAR), Giza, Egypt. www.ijsrp.org International Journal of Scientific and Research Publications, Volume 6, Issue 1, January 2016 409 ISSN 2250-3153 Animals were accommodated under controlled environmental oxide (NO) according to manufacturer’s prescripts using the conditions (23±2°C temperature, 60±10% humidity, 12/12 h corresponding Biodiagnostic colorimetric kits (Cairo, Egypt), light/dark cycle) and were allowed standard chow diet and water while tumor necrosis factor (TNF)-α was assayed using rat TNF- ad libitum. The investigation complies with the Guide for the α Elisa Kit (Enzo life sciences, San Diego, CA, USA). Care and Use of Laboratory Animals (NIH Publication No. 85- Histological examination 23, revised 1996) and was approved by the Ethics Committee of Light microscopic evaluation Faculty of Pharmacy, Cairo University, Cairo, Egypt (Permit The fixed testis was processed for paraffin embedding and 4 Number: PT 922). μm sections were prepared. Testicular sections were stained with Chemicals and drugs haematoxylin and eosin (H&E) and examined using a light Cisplatin was purchased from Mylan (Saint-Priest, France). microscope. Qualitative histopathological damage in the Captopril and telmisartan were obtained from Multipharma seminiferous tubules was graded according to the severity of pharmaceutical and Sigma pharmaceutical industries; degenerative findings: (−) no obvious damage, (+) fewer than respectively (Cairo, Egypt). All other chemicals were of the 25% of seminiferous tubules affected (mild), (++) 25–50% of highest purity and analytical grade. seminiferous tubules affected (moderate), and (+++) over 50% of Experimental design seminiferous tubules affected (severe) (Rezvanfar et al., 2013). Seventy rats were randomly divided into seven groups (10 Histopathological examination of testes was interpreted by an rats each). Group I served as a control. Rats of group II received experienced observer who was blind to the sample identity to orally 0.5 % carboxymethyl cellulose (10 ml/kg) for 15 days. avoid any bias. Animals of groups III & IV received orally captopril dissolved in Immunohistochemical analysis of testicular nuclear factor- water (100 mg/kg) (Fasihi et al., 2012) and telmisartan kappa B (NF-κB) suspended in 0.5 % carboxymethyl cellulose (10 mg/kg) (Fouad Paraffinized testicular sections were rehydrated in xylene and Jresat, 2013) for 15 days; respectively. Those of groups V, and graded ethanol solutions. Samples were heated in citrate VI, and VII received a single intraperitoneal injection of cisplatin buffer (pH 6) for 20 minutes, cooled, and thereafter incubated (10 mg/kg) on the 12th day to induce testicular toxicity (Longo et with primary polyclonal rabbit anti-NF-κB antibody (1:200; al., 2011); whereas, rats of groups VI and VII were treated with Invitrogen, Carlsbad, CA, USA) overnight at 4°C. Sections were captopril and telmisartan; respectively, as previously described. washed with phosphate buffered saline, and incubated for 30 min Four days after cisplatin injection, rats were weighed, at 37°C with biotinylated secondary antibody, and then with anesthetized, and blood samples were collected and centrifuged Avidin DH and biotinylated horseradish peroxidase H complex at 3000 xg for 10 minutes for serum separation to estimate according to Elite ABC kit instructions (Vector Laboratories Inc., testosterone level. Thereafter, animals were euthanized and the Burlingame, CA, USA). After another wash with phosphate reproductive organs were immediately removed, cleaned from buffered saline, the reaction was revealed by diaminobenzidine the adhering tissue, and weighed. The epididymal content for tetrahydrochloride (DAB Substrate Kit, Vector Laboratories Inc., each rat was instantly collected to examine the spermatological Burlingame, CA, USA) and the sections were counterstained parameters. Furthermore, the left testis was fixed in 10% with hematoxylin, dehydrated,