ASSESSMENT OF REPRODUCTIVE FUNCTIONS IN MALE WISTAR RATS TREATED WITH Saccharum officinarum MOLASSES

Abstract

Refined sugar, a major product from Saccharum officinarum, contains mainly sucrose and possesses anti-androgenic effects. Saccharum officinarum Molasses (SOM), a sweet by product obtained during sugar production, is rich in phenolic compounds, minerals and organic acids. The use of SOM as substitute sweetener is increasing because of its nutritional advantage over refined sugar. However, there is paucity of information on its effects on reproductive functions. This study was designed to investigate reproductive functions of SOM-treated male Wistar rats. Saccharum officinarum was obtained from Karu, Nasarawa State and authenticated at the Department of Botany herbarium, University of Ibadan (UIH No.: 22613). Saccharum officinarum juice (SOJ) was obtained using standard procedure. The SOJ was subjected to three cycles of heating and cooling to obtain SOM. The SOM was fractionated with methanol and water and the constituents of fractions (SOMMF and SOMAqF) were identified using GC-MS. Twenty male rats (100-120 g) were grouped into four (n=5) and they received distilled water (1.0 mL/kg/day, Control), SOJ (1.0, 3.2, 10.0 mL/kg/day) orally for 8 weeks, respectively. Another thirty-five male rats (160-180 g) were grouped into seven (n=5) and they received distilled water (1.0 mL/kg/day, Control), SOMMF (1.0, 3.2, 10.0 mL/kg/day) and SOMAqF (0.6, 2.0, 6.4 g/kg/day) orally for 8 weeks, respectively. Testicular and epididymal malondialdehyde were assayed spectrophotometrically. Sperm profile, histology of testes and epididymides were assessed microscopically. Also, testicular cells from twelve rats were isolated, cultured and incubated with SOMMF, SOMAqF, components of SOMMF (Lupeol) and SOMAqF (Diethyl Phthalate). Cell livability, proliferation and testosterone in cultured cells were quantified using ELISA. Data were analyzed using ANOVA at α0.05. The SOMMF (methanol) yielded 72.8% and SOMAqF (aqueous) 27.2%. Major constituents from SOMMF and SOMAqF were lupeol (87.2%) and diethyl phthalate (47.4%), respectively. The SOJ (10.0 mL/kg/day) significantly increased serum testosterone, sperm concentration and abnormal spermatozoa compared to control. The SOJ (10.0 mL/kg/day) significantly decreased sperm livability (38.5±4.5%) compared to iii control (69.7±4.7%). The testicular malondialdehyde 2.4±0.2, 2.6±0.1 U/mg protein of SOMMF (1.0, 3.2 mL/kg/day) and 2.6±0.4 U/mg protein of SOMAqF (6.4 g/kg/day) significantly increased compared to control (1.6±0.2 U/mg protein). The SOMMF (1.0 mL/kg/day) significantly increased epididymal malondialdehyde (43.0±5.2 U/mg protein) relative to control (15.7±6.9 U/mg protein). Similarly, SOMMF (3.2, 10.0 mL/kg/day) significantly decreased sperm livability (79.2±2.4, 71.3±5.0%) compared to control (91.7±2.0%). The SOMAqF (2.0, 6.4 g/kg/day) significantly reduced sperm livability (78.8±2.1, 74.2±3.0%) compared to control (91.7±2.0%). Also, SOMAqF (2.0, 6.4 g/kg/day) significantly increased abnormal spermatozoa (13.1±1.3, 14.2±1.5%) compared to control (9.0±0.7%). Seminiferous tubules and epididymal ducts of SOMAqF-treated rats showed architectural distortion. Testicular cell proliferation significantly increased in SOMMF (2.9±0.3) compared to control (1.9±0.2). Cell livability significantly decreased in Lupeol (1.1±0.2 million/mL) and diethyl phthalate (1.1±0.1 million/mL) compared to control (1.9±0.2 million/mL). In vitro testosterone biosynthesis significantly reduced in diethyl phthalate (0.8±0.1 ng/mL) compared to control (1.2±0.0 ng/mL). Saccharum officinarum molasses caused lipid peroxidation and reduced sperm quality. These actions were linked to its constituents; lupeol and diethyl phthalate. Hence, Saccharum officinarum molasses could adversely affect reproductive functions in male Wistar rats.