Protective Effects of a Polyherbal Combination of Ajuga Bracteosa and Gynura procumbens on Cisplatin-Induced Renal Cytotoxicity Through Modulation of ROS-Mediated Apoptotic Pathways in HEK-293 Cells

Abstract

Cisplatin-induced nephrotoxicity remains a major limitation in chemotherapy, primarily mediated through oxidative stress and apoptosis in renal cells. The present study evaluated the nephroprotective potential of a hydroalcoholic polyherbal extract comprising Ajuga bracteosa and Gynura procumbens (1:1) against cisplatin-induced cytotoxicity in HEK-293 cells. Cytotoxicity was induced using cisplatin (20 µM), while the extract was tested at 25, 50, and 100 µg/mL concentrations following pre-treatment. Cell viability was assessed using the MTT assay, and membrane integrity was evaluated by lactate dehydrogenase (LDH) release. Oxidative stress parameters were analyzed through intracellular reactive oxygen species (ROS) generation, lipid peroxidation (MDA levels), and endogenous antioxidant enzymes (SOD, CAT, and GSH). Mitochondrial membrane potential (ΔΨm) was determined using JC-1 dye, while apoptosis was assessed via caspase-3 activity. Cisplatin exposure significantly reduced cell viability, increased ROS production, induced lipid peroxidation, disrupted mitochondrial function, and elevated caspase-3 activity. Pre-treatment with the polyherbal extract demonstrated a dose-dependent protective effect, restoring cell viability, reducing oxidative stress, preserving mitochondrial integrity, and suppressing apoptosis. The highest extract concentration (100 µg/mL) exhibited effects comparable to the standard antioxidant N-acetylcysteine. The findings suggest that the polyherbal extract confers nephroprotection through modulation of the oxidative stress–mitochondrial apoptosis axis, indicating its potential as a supportive therapeutic strategy against cisplatin-induced renal toxicity.