Methanol Crude Peel Extract of P. granatum Prevents Oxidative Damage in Kidneys of Rats Exposed to Highly Active Antiretroviral Therapy

Abstract

Background

Highly Active Antiretroviral Therapy (HAART) has been linked to oxidative damage to kidney cells leading to renal disease in people living with HIV/AIDS on HAART treatment. The toxic effects of HAART affect the patients’ quality of life leading to poor adherence to their regimen. Therefore, the purpose of this study was to investigate the nephron-protective activity of methanol crude peel extract of Punica granatum (MPEPG) in HAART-administered Wistar rats.

Methods

Thirty male albino Wistar rats weighing between 180–200g were randomly divided into six groups of five rats each. Group one served as normal control and was given distilled water only. Group two serves as a negative control and was given HAART at a dosage of 64 mg/kg. Groups 3 and 4 were given 100 and 400 mg/kg of MPEPG, respectively, while groups 5 and 6 were given MPEPG dosages of 100 and 400 mg/kg along with HAART, respectively, for 40 days. The rats were sacrificed under halothane anaesthesia, and the kidneys were removed for histological evaluation, while blood samples were analyzed for biochemical parameters.

Results

In the HAART (TLD) treated group, there was a significantly high amount of MDA and a lower level of the antioxidant enzymes SOD and CAT. Biochemical analysis revealed that animals treated with HAART (TLD) had significantly higher levels of urea and creatinine, which are biomarkers of kidney damage than the normal control animals. In contrast, all the kidney function markers were returned to normal levels in the HAART-treated group after administration of methanol crude peel extract of P. granatum. The kidney tissues of animals given HAART had considerable structural damage as revealed by histopathological studies. When HAART-exposed rats were treated with MPEPG, both the biochemical and histological results significantly improved.

Conclusion

Methanol crude peel extract of P. granatum provided effective protection against kidney oxidative injury brought on by HAART because of its anti-oxidant and free radical scavenging properties.

Keywords:

• HAART
• oxidative stress
• nephrotoxicity
• Punica granatum

Abbreviations

3TC, Lamivudine; ALB, Albumin; ALP, Alkaline phosphatase; ALT, Alanine aminotransferase; ANOVA; Analysis of variance; ART, Antiretroviral therapy; AST, Aspartate aminotransferase; BIL-T, Total bilirubin; CAT, Catalase; CCl₄, Carbon tetrachloride; DPX, Dibutyl Phthalate in Xylene; DTG, Dolutegravir; HAART, Highly active antiretroviral therapy; HIV, Human immunodeficiency virus; KIU-WC, Kampala International University-western campus; KMnO₄, potassium permanganate; MDA, Malondialdehyde; MPEP, Methanol crude peel extract of P. granatum; OECD, Organization for Economic Cooperation and Development; OS, Oxidative stress; PG, Punica granatum; ROS, reactive oxygen species; SOD, Superoxide dismutase; TCA; trichloroacetic acid; TDF, Tenofovir; TBA, thiobarbituric acid; TLD, Tenofovir, Lamivudine, Dolutegravir; TPro, Total protein.

Data Sharing Statement

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

Ethics Approval and Informed Consent

The study involved the use of animals and the Kampala International University-Western Campus Animal Research Ethics Committee provided ethical approval for all animal experiments (Approval number: SF/202031). 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 of the fewest number of animals possible according to Organisation for Economic Co-operation and Development (OECD) protocols.

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.

Author Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis and interpretation, or in all these areas; took part in drafting, revising, or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.

Disclosure

Allan Wandera reports grants from MAPRONANO ACE, during the conduct of the study. The rest of the authors declare that they have no competing interests.

Additional information

Funding

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