EVALUATION OF WOUND HEALING AND ANTI-INFLAMMATORY PROPERTY OF 5,3’DIHYDROXYFLAVONE AND GENE EXPRESSION OF JAK AND COX-2 IN LIPOPOLYSACCHARIDE-INDUCED RAW264.7 CELL LINE
DOI:
https://doi.org/10.22159/ajpcr.2026v19i1.57182Keywords:
5,3'-dihydroxyflavone, Raw264.7 cell line, MTT, Scratch assay, Nitric oxide assay, Gene expression, Wound healingAbstract
Objectives: The study aims to evaluate the wound healing potential and anti-inflammatory properties of 5,3’-dihydroxyflavone (DHF) and investigate its regulatory effects on the expression of Janus kinase (JAK) and cyclooxygenase-2 (COX-2) genes in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cell line.
Methods: Cytotoxicity was evaluated using the MTT assay. RAW 264.7 cells were treated with DHF (1–100 μg/mL) for 24 h. Absorbance was measured at 490 nm and 630 nm. Wound healing was assessed through a scratch assay. LPS-induced and DHF-treated groups (12.5, 25, 50 μg/mL) were analyzed for scratch closure at 0 and 24 h. Nitric oxide (NO) production was quantified using the Griess assay. Nitrite levels were measured at 520–550 nm. Gene expression of COX-2 and JAK1 was analyzed using quantitative polymerase chain reaction (qPCR), with β-actin as a reference gene. Relative expression was determined by the 2−ΔΔCt method.
Results: The MTT assay confirmed DHF’s non-cytotoxicity, showing ≥90% viability at 1–100 μg/mL (p>0.05). The scratch assay showed enhanced wound closure. At 50 μg/mL, DHF achieved ~75% closure, compared to 45% in the LPS group. NO estimation revealed a dose-dependent reduction. At 50 μg/mL, DHF significantly lowered NO levels (p<0.01). qPCR analysis demonstrated downregulation of COX-2 (45%) and JAK1 (50%) at 50 μg/mL (p<0.001). These findings confirm DHF’s anti-inflammatory and wound-healing potential.
Conclusion: This study demonstrates that DHF enhances wound healing in RAW 264.7 cells by downregulating JAK and COX-2 expression, highlighting its anti-inflammatory potential.
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