الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
Pharmacovigilance was defined by WHO as “the science and activities relating to the detection, assessment, understanding and prevention of adverse effects or any other possible drug-related problems. The American urological association recommended to review the medication history regarding full evaluation for male infertility.Aminoglycoside antibiotics are one of the main bactericidal antibiotic classes commonly used for different types of infection e.g. urinary tract infection, infective endocarditis and blood stream infection.The aim of the present study was to investigate the possible adverse effects of the antibacterial aminoglycoside agents, gentamicin and amikacin, on the male albino rat reproductive system indicated by looking at their effects on reproductive organ weights, sperm parameters and plasma testosterone, FSH, LH, FSH, T3 and T4 levels. Additionally, histopathological examination of the testis was performed. Moreover, the possible explanatory scenarios for the drugs’ adverse effects were examined, including assessment of testicular oxidative stress biomarkers, testicular gene expression of the antioxidant enzymes.To fulfill this aim, the present study was conducted on fifty adult male albino rats aging 8-10 weeks of approximate weights 210-230 g. After acclimatization, animals were randomly divided into five groups; the first and second groups were treated with the therapeutic and double doses of gentamicin respectively (18.25 and 36.5 mg/kg respectively), the third and fourth groups were treated with the therapeutic and double doses of amikacin (54.75 and 109.5 mg/kg, respectively); while the fifth group served as control and was treated with only the vehicles of the drug preparations. All drugs were administered intraperitoneally once daily for 14 days.Just before sacrificing, blood samples were collected from the retro-orbital venous plexus by means of heparinized capillary tubes. The collected blood was received into heparinized sampling tubes and centrifuged at 1500 xg for 15 minutes, then the clear plasma samples were collected in sterile Eppendorf tubes. Plasma samples were kept frozen (-20 °C) till used for measuring the reproduction related hormones using ELISA. Sacrifice by decapitation was made at two time points for 5 rats in each group; the first sacrifice was on the 15th day and second sacrifice was on the 60th day from the start of the treatment to determine the possible short- and long-term adverse effects on reproductive system. Additionally, to assess the reversibility of the drug related adverse effects.Total body weight and male reproductive organ weights were measured immediately after sacrifice using digital balance. Sperm motility and count were examined microscopically with light microscope at 400x magnification while sperm viability and abnormalities were determined microscopically under magnification of 400x and 1000x. Hemocytometer was used for sperm counting. One testis was dissected out, part of which was frozen at – 80oC for the purpose of gene expression assay using real-time PCR. The other part was homogenized with phosphate buffer solution at PH 7.4 and centrifuged at 10000 rpm for 10 minutes at 4oC. The supernatant was kept at -20oC until used in determining testicular oxidative stress biomarkers using spectrophotometry. The other testis was preserved in formalin 10% for the purpose of histopathological examination. After 24 hours, the testes were prepared for staining and examination. The following results were obtained:1-Male reproductive organ weight effects:A-On the 1st sacrifice day, ventral prostate weight wasn’t affected with either gentamicin or amikacin. Epididymis weight was decreased with the two doses of gentamicin and the double dose of amikacin. Seminal vesicle weight was decreased with the two doses of gentamicin and the double dose of amikacin. Testis weight was reduced with the two doses of gentamicin and amikacin.B-On the 2nd sacrifice day, ventral prostate and seminal vesicle weight weren’t affected with either gentamicin or amikacin. Epididymis weight was decreased with the two doses of gentamicin and amikacin. Testis weight was reduced with the double dose of gentamicin only.2-Sperm parameter effects:A-On the 1st sacrifice day, sperm count and progressive motility were reduced with the two doses of gentamicin and the double dose of amikacin. Sperm viability was reduced with the double dose of gentamicin and amikacin. Sperm tail abnormalities were increased with the two doses of gentamicin and the double dose of amikacin. Sperm head abnormalities were increased with the double dose of amikacin only.B- On the 2nd sacrifice day, sperm count and viability were reduced with the two doses of gentamicin and amikacin. Sperm progressive motility was decreased with the two doses of gentamicin and the double dose of amikacin. Sperm tail abnormalities were increased with the two doses of gentamicin and amikacin. Sperm head abnormalities were increased with the double dose of amikacin only. 3- Plasma hormone levels effects:A-On the 1st sacrifice day, serum testosterone level was reduced with the two doses of gentamicin, but it wasn’t affected with amikacin treatment. Plasma FSH and LH level were increased with the therapeutic and double dose of gentamicin and amikacin. Plasma levels of TSH, T3 and T4 have not been affected.B-On 2nd sacrifice day, serum level of testosterone, FSH and LH hormone returned to normal where the difference between the groups wasn’t significant. Serum levels of TSH, T3 and T4 weren’t affected.4-Testicular oxidative stress biomarker effects:A-On the 1st sacrifice day, testicular MDA concentration was increased with the therapeutic and double dose of gentamicin and amikacin. Testicular TAC and testicular SOD and catalase enzyme activity were reduced with the therapeutic and double dose of gentamicin and amikacin.B-On the 2nd sacrifice day, the testicular oxidative stress has not been observed in the treated groups, where the groups treated with the therapeutic and double dose of gentamicin and amikacin showed normal values of oxidative stress biomarkers relative to control.5- Testicular anti-oxidant gene expression effects:A-On the 1st sacrifice day, testicular catalase gene expression was reduced with the double dose of gentamicin only. Testicular SOD gene expression was reduced with the double dose of amikacin only testicular glutathione peroxidase gene expression was reduced with the double dose of gentamicin and amikacin.B- On the 2nd sacrifice day, gene expression of catalase, SOD and glutathione peroxidase enzyme returned to normal where the difference between the groups wasn’t significant.6-Histopathological effects:A- On the 1st sacrifice day, therapeutic dose gentamicin showed variable degree of degenerative changes of the lining epithelial cells of some seminiferous tubules was seen with incomplete spermatogenesis. Double dose gentamicin showed marked necrosis of spermatogenic cells in seminiferous tubules that lined by few layers of degenerated germ cells and distension of the lumen of some tubules with necrotic cells. In specimens from animals treated with the therapeutic dose of amikacin, most of the seminiferous tubules were compact with each other and the spermatogenic layers appeared somewhat normal. While in those treated with the double dose of amikacin, degeneration of germinal epithelium and loss of spermatids with incomplete of spermatogenesis was observed.B- On the 2nd sacrifice day, most of testicular cells and structures of animals treated with therapeutic or double dose of gentamicin or amikacin returned to normal relative to control group.Depending on the above mentioned results, the present study shed the light on the possible risk of using gentamicin and amikacin on the male reproductive system. Gentamicin, and to a lesser extent amikacin, might represent a hazard on male fertility concluded by the drug adverse effects on reproductive organ weights and testicular tissue structure as well as sperm parameters. These effects were associated with androgenic hormone deficiency and testicular oxidative stress indicated by alteration in levels of oxidative stress biomarkers and their gene expression, therefore, monitoring of male reproductive aspect in patients receiving these drugs is required and appropriate antioxidants are recommended while using these drugs.