Antioxidant Effect of Lemongrass (Cymbopogon citratus) Extract on Paraquat-Induced Oxidative Stress in Sheep Red Blood Cells Using In-vitro Phytochemical Analysis

Abstract

Aims: Oxidative stress, which causes harm to cells, is a significant factor in many diseases. Red blood cells are easily damaged by free radicals. Lemongrass has powerful abilities to fight these free radicals’ because it contains a lot of phytochemicals. The study examined how effective lemongrass is in protecting red blood cells from damage caused by Paraquat. Additionally, phytochemicals present in lemongrass are also analysed. Methodology:  The preparation of lemongrass leaf extract involved drying and grinding the leaves, followed by extraction with acetone using the Soxhlet method. Phytochemical analysis was done to reveal the presence of alkaloids, saponins, flavonoids, tannins, phenols, and acids. A 10% RBC suspension was prepared from sheep blood. Oxidative stress was induced in RBCs using Paraquat, Ascorbic acid, and the extract. Antioxidant analysis included protein concentration via the Lowry method, Thiobarbituric acid reactive substance (TBARS) estimation for lipid peroxidation, Glutathione (GSH) estimation, and Superoxide dismutase (SOD) activity assay. Statistical analysis was performed using GraphPad Prism, with significance set at p<0.05. Results: The phytochemical analysis revealed the presence of flavonoids, terpenoids, saponin, and tannin in lemongrass, while no acids or alkaloids were detected in the plant. In Group II exposed to Paraquat toxicity, total protein decreased significantly compared to the control group (10.24±0.53 mg/dl to 6.3±0.49 mg/dl). TBARS activity notably increased in Paraquat-treated group (629.7±2.85) compared to control (398.8±3.51), while Lemongrass and ascorbic acid treatments showed significant decreases (513.05±4.54 and 504.01±1.97, respectively). SOD and GSH levels decreased significantly in Paraquat-treated group (6.08±0.4 and 80.86±4.9 µg/mg of protein) compared to control (11.13±1.1 and 109.81±8.6, respectively), but Lemongrass treatment increased levels (14.12±2.0 and 114.8±4.5 µg GSH/mg of protein). Lemongrass demonstrated pronounced protective effects against oxidative stress, normalizing SOD levels and increasing GSH levels, comparable to ascorbic acid treatment. Conclusion: The study successfully induced oxidative stress in RBCs with Paraquat, presenting avenues for further exploration in understanding cellular responses. Additionally, the research highlights Lemongrass's significant preventive impact (100μg), possibly attributed to its bioactive phytochemicals, including flavonoids, phenols, saponins, and tannins. This underscores Lemongrass's potential in countering oxidative damage induced by Paraquat in RBCs.