Protective Effects of Moringa oleifera Aqueous Leaf Extract on TM3 Leydig Cells Against Hydrogen Peroxide-Induced Oxidative Stress

Abstract

Introduction

Moringa oleifera (M. oleifera) is a medicinal plant rich in dietary antioxidants that may protect reproductive cells from oxidative stress-induced damage and male fertility problems. This study investigated M. oleifera aqueous leaf extracts (ALE) effects on TM3 Leydig cells exposed to hydrogen peroxide (H₂O₂).

Methods

M. oleifera ALE and ascorbic acid (10−1000 µg/mL) were assessed for antioxidant activity. TM3 Leydig cells were exposed to various concentrations of M. oleifera ALE (0−1000 µg/mL) or α-tocopherol (50 µM) for 24 h, alongside 200 µM H₂O₂, with or without hCG (4 ng/mL). Cell viability, testosterone production, mitochondrial membrane potential (MMP), total protein and oxidative stress parameters (reactive oxygen species [ROS], superoxide dismutase [SOD], reduced glutathione [GSH], total antioxidant capacity [TAC] and lipid peroxidation [MDA]) were assessed.

Results

Ascorbic acid demonstrated superior DPPH radical scavenging and FRAP activity compared to M. oleifera ALE at most concentrations (p  < 0.0001). H₂O₂ (200 µM) exposure significantly reduced cell viability, MMP, SOD, GSH and TAC levels, while increasing ROS and MDA levels (p  < 0.05). Treatment with M. oleifera ALE or α-tocopherol significantly increased cell viability, total protein, SOD activity and GSH levels while reducing ROS and MDA compared to H₂O₂ control (p  < 0.05). However, testosterone levels remained unchanged (p  > 0.05).

Conclusions

M. oleifera ALE effectively protects the TM3 Leydig cells from H₂O₂-induced oxidative stress by restoring the redox balance, though unchanged testosterone levels suggest limitations in using TM3 Leydig cells for assessing testosterone synthesis.