Testicular structure following interaction of alcohol with antiretroviral therapy: Role of Virgin coconut oil extract in an experimental animal model.

Abstract

The consumption of alcohol by people living with HIV/AIDS is associated with a graver prognosis. Long term use of antiretrovirals has known health challenges that may be aggravated by concomitant alcohol use. This study investigated Virgin coconut oil (VCO) as an adjuvant to the deleterious effects of combining highly active antiretroviral therapy (HAART) and alcohol on the cyto-architecture and functions of the testis in an experimental animal model. The study was conducted in two phases – phase one involving HAART and VCO administration and phase two involving ethanol, HAART and VCO co-administration. In phase 1, twenty adult male Sprague Dawley rats weighing between 153g to 169g were divided into four groups with five animals per group and forty adult male SpragueDawley rats weighing between 165g to 176g, were used in phase 2 and divided into eight groups with five animals per group. The animals were subjected to various treatments with HAART, Ethanol and VCO according to the protocol. Blood was collected for hormonal (LH, FSH, Testosterone) and antioxidant marker (MDA and GSH) assays. Epididymal seminal fluid was analysed for sperm concentration, motility and morphology. The testes were examined for histopathological and histochemical changes using H&E, PAS and Gordon and Sweet’s silver stains. Testicular ultrastructure was examined using transmission electron microscopy. Morphometric measurements were done on the seminiferous tubules examining seminiferous tubular diameter, epithelial and basement membrane thickness, Leydig cell diameter and nuclear volume. Stereological studies were carried out examining volume densities and absolute volumes of the germinal epithelium, lumen and the intersitium. Results of the semen parameters showed a significant decline in sperm counts (P<0.01) and motility (P<0.05) in animals treated with HAART alone, ethanol alone or with HAART + Ethanol when compared to the control animals. The groups receiving adjuvant VCO + ethanol also had significantly increased sperm counts (P<0.05) and sperm motility (P<0.01) than ethanol alone. Likewise the group receiving VCO + ethanol + HAART showed significantly increased sperm counts (P<0.05) and sperm motility (P<0.01) than ethanol + HAART alone. Hormonal assay indicated a significant increase in testosterone levels relative to the control animals (P<0.01). There was a significant increase in FSH levels of VCO + ethanol + HAART relative to the ethanol + HAART treated animals (P<0.01). Testicular GSH level was significantly decreased (P<0.05) in the ethanol alone treated group. A significant increase (P<0.01) was also observed in the VCO + ethanol and VCO + ethanol + HAART. Changes in the LH and MDA levels were not statistically significant among all treated animals. Histo-morphological studies showed HAART caused some damage to seminiferous tubular architecture with a significant decline in epithelial height closely mirrored by extensive abnormal reticulin framework and presence of many positive PAS cells. Testis ultrastructure of HAART-treated animals also showed significant increase (p<0.01) in basement membrane thickness with decrease in Leydig cell nuclear diameter (p<0.05) and volume (p<0.01) when compared to controls.

Mitochondrial cristae appeared collapsed and Sertoli cells showed cytoplasmic vacuolations. In addition, the use of ethanol alone and ethanol + HAART showed extensive degeneration in the seminiferous tubular epithelium, disorganized basement membrane and widened, hypocellular interstitium. Adjuvant treatment of VCO with HAART, ethanol, or ethanol + HAART showed improved testicular morphology, reversed HAART and ethanol histopathology as well as improved HAART-induced testicular ultrastructural alterations. In conclusion, the use of Virgin coconut oil was found to mitigate the deleterious effects of ethanol and HAART thereby significantly preserving and promoting testicular function and fertility. Keywords: Alcohol, Virgin coconut oil, HAART, Ultrastructure, Testis