Comparative Study of the Phytochemistry and Anti-hemorrhoidal Activity of Hydroethanolic Extracts of Persea americana Mill. Seeds and Nephrolepis biserrata

Abstract

Background: Hemorrhoids are a common condition affecting many adults and significantly reduce quality of life, while conventional treatments are often costly and less accessible. This has led to growing interest in plant-based alternatives like Nephrolepis biserrata and Persea americana seeds, which show anti-inflammatory potential but lack sufficient direct clinical evidence. Aims: This study aims to compare the anti-hemorrhoidal efficacy of Persea americana seed extracts (SPA) and Nephrolepis biserrata (NB) in rats by evaluating inflammatory, hepatic, and morphological parameters. Study Design:  A randomized experimental study in rats with induced hemorrhoids compared the anti-inflammatory, hepatoprotective, and clinical effects of Persea americana seeds and Nephrolepis biserrata extracts obtained via 30/70 water-ethanol extraction. Place and Duration of Study: Chemical and biological analyses were conducted at LaKReCA (UNSTIM) and URMAPha (University of Abomey-Calavi), in Republic of Benin, between January and December 2025. Methodology: the phytochemical, antioxidant, and anti-hemorrhoidal properties of Persea americana seeds. The hydroethanolic extract (Water/Ethanol: 30/70) was obtained by maceration. Qualitative and quantitative phytochemical analyses were conducted to identify secondary metabolites. Biologically, the parameters measured after 5 days of treatment in Wistar rats included total protein levels (protidemia), erythrocyte sedimentation rate, ASAT and ALAT levels, as well as variations in the recto-anal coefficient. Results: Nephrolepis biserrata (NB) shows a higher extraction yield (16.36%) than Persea americana seeds (SPA) (14.36%), while SPA contains more total polyphenols (289.6 mg gallic acid equivalents per gram versus 75 mg). However, NB has a higher flavonoid content (38.5 mg quercetin equivalents per gram versus 35.76 mg) and stronger antioxidant activity, with a half maximal inhibitory concentration of 9.13 × 10⁻³ mg/mL compared to 1.93 × 10⁻² mg/mL for SPA. In treated rats, NB at 250 mg/kg significantly improved inflammatory and biochemical parameters, reducing erythrocyte sedimentation rate from 4.66 to 2, stabilizing serum proteins, and improving liver enzymes (alanine aminotransferase 88.7; aspartate aminotransferase 172.19). It also showed better recto-anal coefficient normalization (0.001558) than SPA. Overall, NB at 250 mg/kg was the most active fraction, although SPA at 500 mg/kg showed partial hepatoprotective effects. Conclusion: Both extracts exhibit significant anti-inflammatory and antioxidant properties, but NB at 250 mg/kg demonstrates the best overall therapeutic efficacy.