In-vitro Anti-inflammatory Effect of Caesalpinia sappan (Kayu Secang/Sappanwood) Extract on LPS-induced RAW264.7 Cells

Abstract

Inflammation acts as the body's defense mechanism against infections, tissue injury, and cellular stress, however, prolonged inflammation can contribute to the development of chronic diseases. Caesalpinia sappan is a medicinal plant known to contain bioactive compounds, particularly brazilin, with potential anti-inflammatory properties. This study aimed to evaluate the anti-inflammatory activity of Caesalpinia sappan extract (CSE) in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. FTIR spectroscopy was used to characterize the extract, while cytotoxicity was assessed using the WST-8 assay. The production of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and Nitric Oxide (NO) was measured using ELISA and Griess assays, respectively. Gene expression of Tnf-ɑ, Il-6, and Nos2 was determined using Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR). FTIR analysis revealed the presence of functional groups associated with phenolic compounds. CSE concentrations ranging from 10–30 μg/mL maintained cell viability above 70%, indicating low cytotoxicity, with no significant effect on cell viability. Furthermore, CSE significantly reduced TNF-α, IL-6, and NO production (p < 0.05), with the greatest inhibitory effect observed at 30 μg/mL. RT-qPCR analysis also demonstrated downregulation of Tnf-ɑ, Il-6, and Nos2 expression following CSE treatment. These findings suggest that CSE possesses anti-inflammatory potential by suppressing pro-inflammatory gene expression and reducing inflammatory mediator production in LPS-induced RAW264.7 cells.