Cardiovascular toxicity of combined administration of ethanol and combination antiretroviral therapy in HIV naïve diabetic Sprague Dawley male rats

Abstract

Atripla, a combination antiretroviral drug (cART) was the first once daily antiretroviral treatment to be approved by FDA in 2008. Since then, Atripla has been the first line of treatment in HIV/AIDS management, and one of widely used medications, with 76% of people living with HIV on medication. The incidence of diabetes in Africa has increased, owing to decreased mobility and increasing urbanization in the continent. Excessive alcohol consumption on the continent, especially in South Africa is common across all age groups, with ethanol (alcohol) often taken concomitantly with antiretroviral drugs. This results in co-presence of cART and alcohol in a diabetic condition. Adult male Sprague-Dawley rats divided into 8 groups were used: untreated (negative control), diabetic (diabetes, diabetes + alcohol, diabetes + Atripla, and diabetes + Alcohol + Atripla), and treated (alcohol alone, alcohol and Atripla, and Atripla alone). Harvested hearts’ morphology was investigated (surface area and weight) before being processed and histological investigations were carried out (H&E for structural differences, Pentachrome for collagen and muscle profile, and Verhoeff Van Gieson for collagen and elastic fiber profiling). Fasting glucose levels were measured and recorded throughout the study. Enzyme-linked immunosorbent assays for troponin I&T (myocardial degeneration), MDA, and GPx (oxidative stress) were performed. Additionally, immunohistochemistry for expression of eNOS (as a marker for endothelial homeostasis) and c- reactive protein CRP (as a marker for inflammation) were performed. The fasting glucose was significantly increased in diabetes + Alcohol + Atripla group (DBALCARV) throughout the study period (p=0.001), and morphological examination revealed a decrease in body weight (p=0.001) and total surface area of the heart (p=0.01). The coronary 5 artery had a significant decrease in tunica media thickness (p=0.02), while the aorta was increased (p=0.001). The collagen in the coronary arteries was significantly increased when examined with Pentachrome special stain (p=0.032). The collagen was also significantly increased in both ventricle (p=0.015) and atrium (p=0.013). Qualitative analysis of c-reactive protein and endothelial nitric oxide synthase immunohistochemistry showed a reduction in localization of both antibodies. In conclusion, combination of cART, diabetes, and alcohol resulted in changes in cardiac and vascular morphology, fasting glucose, body weight, oxidative stress, myocardial degradation markers, and collagen expression.