EFFECTS OF QUASSIA AMARA AND QUASSIN ON EPIDIDYMAL FUNCTIONS AND HORMONAL PROFILES IN RATS AND THEIR OFFSPRINGS

Abstract

Quassia amara is a medicinal plant with antimicrobial and bitter stomachic properties. This plant and its bioactive compound quassin, have been reported to decrease sperm count, sperm motility, male fertility index and alter sperm morphology. However, information on the effects of Q. amara and quassin on epididymal functions such as sperm capacitation, acrosome reaction and epiditymal proteins is scanty. The effects of Q. amara and quassin on epididymal functions in Wistar albino rats were therefore investigated. Quassia amara was obtained from University of Ibadan and authenticated at Forest herbarium, lbadan. Pulverized stem bark was extracted with methanol and concentrated using rotary evaporator, Q. Amara extract (100 mg/kg), quassin (0.1 and 2 mg/kg) and distilled water (0.5 ml) were administered by gavage (p .o) daily for 6 weeks to male rats (180-200g. n=5), and thereafter sacrificed. Sperm motility, viability and sperm count were examined microscopically. Serum from each rat was analysed for Follicle Stimulating hormone (FSH), Luteinizing Hormone (LH) and testosterone by enzyme immunoassay technique alanine Aminotransferase (ALT), Aspartale Aminotransferose (AST) and Alkaline Phosphatase (ALP) were measured spectrophotometrically. Sperm capacitation and acrosome reaction were determined by Coomassie brilliant blue staining technique. Epididymal hongrESI protein expression was assessed using immunohistochemical technique. After five weeks or treatment, female rats were cohabited with the male rats for seven days. Morphometric valiables of offspring were measured on day one of parturition wing digital Vernier caliper. Data were analysed using Student's t- lest and ANOVA at p=0.05. Quassia amara (100 mg/kg) significantly decreased sperm motility (93.8±1.0 vs 42.0±10.2)%. viability (97.31±0.8 vs 88.0±2.6)% and count (1212±4.2 vs 58.0±4.2)million/mL relative to controls. Similarly, quassin (0.1 mg/kg) significantly decreased sperm motility (93.8±1.0 vs 68.0±3.7)%, viability (97.3±0.8 vs 88.6±3.4)% and count (121.2±4.2 vs 94.0±7.9)million/mL . Q. amara significantly reduced LH (1.5±0.2 vs 0.6±0.1)mlU/ml, testosterone (0.3±0.1 vs 0.1±0.03)ng/mL. Similarly, quassin significantly reduced LH-(1.510.2 vs 0.7±0.1)mlU/mL. testosterone (0.3±0.1 vs 0.1±0.03)ng/mL and FSH (4.4±0.5 vs 1.6±0.2)mlU/mL. There was epithelial derangement of the epididymis and seminal vesicles, but not the testes. The ALT (11.8±3.8 vs 18.13±2.7)U/L, AST (14.0±1.8 vs 12.2±1.5)U/L, ALP (144.6±19.9 vs 92.4±15.0)U/L, cholesterol (1.4±0.1 vs 1.3±0.1)mmol/L, triglyceride (0.5±0.1vs 0.4±0.1)mmol/L, LDL (0.7±0.1 vs 0.7±0.1)mmol/L, and HDL (03±0.1 vs 0.6±0.1)mmol/L, were not affected by Q. amrara. The percentage of capacitated acrosome reacted sperm was significantly lower in Q. Amara (95.2.±0.9 vs 27.2±12.0) and quassin (95.2±0.9 vs 24.0±2.7) treated rats. Total epididymal protein was significantly reduced in Q. amara (1.7±0 .3 vs 0.7±0 .1)g/dL. and quassin (1.71±0.3 vs 0.6±0.1)L while total testicular protein was significantly increased by Q. amara (0.9±0.1 vs 1.4±0.2)g/dL. and quassin (0.910.I vs 1.310.1)g/dL. Epididymal hongrESI protein expression was repressed by treatment with Q amara and quassin. Fertility was zero in 80% of treated rats, and a reduction in anogenital distance (0.3±0.01vs0.2±0.02)cm was observed in male offspring of the rats. Quassia amara and quassin suppressed epididymal hongrESI protein expression and decreased the levels of reproductive hormones in the rats. Decline in anogenital distance of the offspring suggests Quassia amara action may be transgenerational Keywords: Quassia amara, Quassia, Sperm capacitation, Aerosome reaction. Epididymal protein Word count: 491