Past Issue

Volume 13, Number 1, Apr-Jun 2019 Pages: 57-65

The Effects of Olive Leaf Extract on The Testis, Sperm Quality and Testicular Germ Cell Apoptosis in Male Rats Exposed to Busulfan

Sepideh Ganjalikhan Hakemi, M.Sc, 1, Fariba Sharififar, Ph.D, 2, Tahereh Haghpanah, Ph.D, 1, *, Abdolreza Babaee, M.Sc, 1, Seyed Hassan Eftekhar-Vaghefi, Ph.D, 1, 3, *,
Department of Anatomy, Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran
Herbal and Traditional Medicines Research Center, Department of Pharmacognosy, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman, Iran
Department of Anatomy, Kerman Branch, Islamic Azad University, Kerman, Iran
*Corresponding Address: P.O.Box: 76169-14115 Department of Anatomy Afzalipour Faculty of Medicine Kerman University of Medical Sciences Kerman Iran,



Busulfan (BU) has a destructive effect on the male reproductive system. The goal of this study was to assess the effects of olive leaf extract (OLE) as a source of antioxidants and phenolic compounds, on BU-induced damages in rat testes.

Materials and Methods

In this experimental study, 40 male Wistar rats were randomly divided into 5 groups. The control group (CTL) received a single intraperitoneal (i.p.) injection of dimethyl sulfoxide (DMSO), followed by oral administration of distilled water for 5 weeks. In BU group, BU (10 mg/kg) was administrated i.p. once. In co- treatment groups, first, received BU (10 mg/kg, a single i.p. injection) then, OLE was administrated orally at different doses of 250 mg/kg (BU+OLE 250), 500 mg/kg (BU+OLE 500) and 750 mg/kg (BU+OLE 750), for 5 weeks. Next, blood and sperm samples were collected. The left testis was removed to investigate testicular parameters and apop- tosis by using H&E and TUNEL staining, respectively. All data were analyzed by SPSS software and a P<0.05 was considered significant.


There was a significant decline in sperm viability (P=0.017), number of primary spermatocyte (PS) (P=0.001) and Leydig cells (P=0.023) in the BU group versus the CTL group. OLE at three doses could repair these defects ver- sus BU group. Increases in apoptotic spermatogonia cells (SG) due to BU were significantly reduced by OLE 250 and 500 mg/kg (P<0.01). A reduction in germinal epithelium height and an increase in apoptotic SG were observed in BU+OLE 750 group vs. other groups (P<0.01) and alkaline phosphatase (ALP) was at the highest level, also Aspartate aminotransferase (AST) increased markedly vs. CTL (P=0.024).


Oral administration of OLE at the doses of 250 and 500 mg/kg could be helpful in ameliorating BU- induced toxicity in rat testes, while OLE 750 mg/kg not only did not cause positive effects, but also could exacerbate the harmful effects.