Assessment of Hepatorenal Protective Effects of Trigonella foenum-graecum and Hibiscus rosa-sinensis Extracts in Phenylhydrazine-Induced Anemic Mus musculus

Abstract

Anaemia induced by oxidative agents like phenylhydrazine (PHZ) is often accompanied by hepatic and renal dysfunction due to increased oxidative stress and free radical generation. This study assessed the hepatoprotective and nephroprotective effects of Hibiscus rosa-sinensis (China rose) and Trigonella foenum-graecum (fenugreek) leaf extracts in PHZ-induced anaemic mice over a 60-day period. Mice treated with PHZ exhibited significantly elevated serum liver biomarkers—ALT (86.5?±?3.6 U/L), AST (120.3?±?4.2 U/L), ALP (210.7?±?7.5 U/L), total bilirubin (1.85?±?0.15 mg/dL), and direct bilirubin (0.75?±?0.05 mg/dL)—along with decreased albumin levels (2.3?±?0.2 g/dL), indicating hepatocellular damage. Kidney function parameters also showed marked derangement with increased serum creatinine (1.52?±?0.08 mg/dL), blood urea nitrogen (BUN) (46.3?±?3.1 mg/dL), and uric acid (5.8?±?0.3 mg/dL). Treatment with China rose and fenugreek extracts, individually and in combination, significantly restored liver and kidney markers toward normal levels, with the combination group showing the most effective recovery (e.g., ALT: 42.1?±?2.1 U/L; creatinine: 0.84?±?0.05 mg/dL). The outcomes were comparable to those observed with standard ferrous sulfate treatment. These findings demonstrate the potent hepato-renal protective effects of China rose and fenugreek extracts, likely mediated through their antioxidant properties, highlighting their potential as natural therapeutic agents in anaemia management.

Introduction

Anaemia, a widespread global health problem, occurs due to reduced hemoglobin or red blood cells, leading to poor oxygen transport. Iron-deficiency anaemia is the most common type, especially in developing countries. Although pharmaceutical iron supplements are effective, they may cause side effects such as gastrointestinal discomfort and oxidative stress, prompting interest in herbal alternatives.

Phenylhydrazine (PHZ) is commonly used in research to induce hemolytic anaemia in animals by damaging red blood cells through oxidative stress. PHZ also causes liver and kidney toxicity, making it useful for testing the protective effects of medicinal plants.

Two medicinal plants—Hibiscus rosa-sinensis (China rose) and Trigonella foenum-graecum (fenugreek)—are known for strong antioxidant, hepatoprotective, nephroprotective, and hematopoietic properties due to their rich phytochemical content. Previous studies show that extracts of these plants can improve blood parameters and protect organs from oxidative damage.

Purpose of the Present Study

The study aimed to investigate:

• Subacute toxicity

• Anaemia recovery potential

• Liver and kidney protective effects

of Hibiscus and Fenugreek leaf extracts, individually and in combination, in PHZ-induced anaemic mice over 60 days, compared with standard ferrous sulfate.

Methods Overview

• Plant materials (Hibiscus bark and Fenugreek seeds) were extracted using water and petroleum ether.

• Swiss albino mice were divided into control, plant-treated, anaemia-induced, and combination-treatment groups.

• Acute and sub-acute toxicity studies established safe doses (400 mg/kg and 800 mg/kg).

• Anaemia was induced using phenylhydrazine (10 mg/kg).

• Hematological and biochemical (kidney and liver function) parameters were evaluated at multiple time points.

Key Findings – Kidney Function

PHZ-induced anaemic mice showed:

• High BUN, high creatinine, and high uric acid, indicating kidney damage.

• BUN increased from 9.6 mg/dL (control) to 27.4 mg/dL (anaemia).

• Creatinine increased from 0.35 mg/dL to 1.0 mg/dL.

• Uric acid increased from 0.60 mg/dL to 1.71 mg/dL.

These changes confirm PHZ-induced oxidative renal injury.

Effect of Plant Extracts

• Both Hibiscus and Fenugreek reduced kidney damage markers.

• The combined extract (400 mg/kg each) showed the strongest improvement, nearly matching the standard ferrous sulfate.

• By Day 60, combination treatment reduced BUN to 15.92 mg/dL, compared to 29.39 mg/dL in untreated anaemic mice.

• Creatinine and uric acid levels also significantly decreased under combined treatment.

Conclusion

Hibiscus rosa-sinensis and Trigonella foenum-graecum extracts demonstrate:

• Strong antioxidant activity

• Effective nephroprotective (kidney-protecting) effects

• Beneficial improvement of anaemia

• Best results when used together

They may serve as safer plant-based alternatives or supplements to conventional iron therapy for managing anaemia and related oxidative organ damage.

Conclusion

The present study demonstrates that Hibiscus rosa-sinensis (China rose) and Trigonella foenum-graecum (fenugreek) leaf extracts exhibit significant hepatoprotective and nephroprotective effects in phenylhydrazine-induced anemic mice. The extracts effectively ameliorated elevated liver enzymes (ALT, AST, ALP), bilirubin levels, and kidney function biomarkers (creatinine, BUN, uric acid), indicating restoration of hepatic and renal function. Among the treatment groups, the combination of both extracts showed the most pronounced recovery, comparable to standard ferrous sulfate therapy. These protective effects are likely attributed to the antioxidant potential of the plant extracts. Thus, China rose and fenugreek may serve as promising natural adjuvants for managing anemia-associated organ toxicity.

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Copyright © 2025 Seema Dixit, Nisha Uraiti. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.