Hepatoprotective effect of methanol fruit extract of Punica granatum L in highly active antiretroviral therapy-induced toxicity in Wistar rats

Abstract

Prolonged use of Highly Active Antiretroviral Therapy (HAART) has been linked to toxicity, particularly hepatotoxicity. There are few effective drugs for HAART patients that promote hepatic cell regeneration and prevent liver injury. Therefore, the purpose of this study was to investigate the hepato-protective activity of Methanol fruit extract of Punica granatum (MFEPG) in HAART-administered rats. Thirty rats weighing between 150–200 g were randomly divided into six groups and each group comprised of five rats. Distilled water was given to the rats in group one. Only HAART was given to the rats in group two. MFEPG at doses of 100 and 400 mg/kg was given to the rats in groups three and four. MFEPG dosages of 100 and 400 mg/kg along with HAART were given to the rats in groups five and six, respectively. All treatments were via oral gavage daily for 40 days. Under halothane anesthesia, all rats were sacrificed on day 41. Liver tissues were utilized for lipid peroxidation marker; Malondialdehyde (MDA), antioxidant enzymes; Superoxide dismutase (SOD) and Catalase (CAT) and histological evaluation, while blood samples were examined for biochemical parameters (AST, ALT, ALP, Total cholesterol, Total protein, and Albumin). The HAART-treated group exhibited a significantly higher amount of the lipid peroxidation end product; MDA, and significantly lower levels of antioxidant enzymes; SOD, and CAT. Liver enzymes and total cholesterol were significantly increased with a significant reduction in Total protein and Albumin levels in the HAART-treated group. Conversely, the liver function biomarkers were returned to normal levels in the HAART and MFEPG-treated groups. Histopathological studies revealed that when HAART-exposed rats were treated with MFEPG, both the biochemical and histological results significantly improved. Thus, the antioxidant activity of MFEPG provides protection against HAART-induced liver oxidative damage. More research is needed to determine the safety of using MFEPG in humans.

Keywords:

• HAART
• oxidative stress
• hepatotoxicity
• Punica granatum
• liver biomarkers of damage

Acknowledgments

We are thankful to the Central Diagnostic Laboratory, College of Veterinary Medicine Animal Resources and Biosecurity, Makerere University and Biochemistry Laboratory, Kampala International University for availing laboratory space.

Ethical considerations

Ethical Approval No.: SF/202031 was granted by the Kampala International University Animal Research Ethics Committee prior to the research. All procedures and animal work were carried out in accordance with the guide for the care and use of laboratory animals. The animals were treated humanely to avoid any discomfort, such as excessive pain and stress. The study adhered to the 3Rs principles of Reduce, Refine, and Replace to ensure use the fewest number of animals possible according Organization for Economic Cooperation and Development (OECD) protocols.

Authors’ contributions

This work was carried out in collaboration between all authors. K.S., E.W., and C.K., conceptualized and designed the study. E.K., M.P., A.W., J.M., and R.S. executed the laboratory work. K.S., and E.K. analyzed the data and wrote the first draft of the manuscript and managed manuscript revisions. All authors read and approved the final manuscript.

Disclosure statement

The authors declare that they have no competing interests.

Data availability

All data generated by this study have been submitted with this manuscript.

Additional information

Funding

The authors acknowledge with sincere gratitude the funding from Africa Center of Excellence in Materials Product Development and Nanotechnology (MAPRONANO)-Makerere University (Project ID: P151847).