In vitro Anti-inflammatory and Anti-allergenic Effects of Thai Herbal Formula and its Isolates, Rhinacanthin-C and Rhinacanthin-N in RBL-2H3 Cells and RAW264.7 Macrophages
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Abstract
Herbal remedies are an alternative treatment for allergic skin inflammation, reducing reliance on conventional chemical orthodox drugs which may have potential risks and side effects. This study evaluated the effects of a Thai herbal formulation extract (HFE) on interleukin 4 (IL-4) production in RBL-2H3 cells, as well as its inhibitory effect on nitric oxide, tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), in RAW264.7 cells. Phytochemical analysis and high-performance liquid chromatography (HPLC) were also performed to identify the active compounds in HFE, which was prepared by maceration in 95% ethanol. IL-4 inhibition in RBL-2H3 cells was measured using an enzyme-linked immunoassay (ELISA). Nitric oxide inhibition was assessed with the Griess reagent, while TNF-α and IL-6 levels were also measured by ELISA. Cytotoxicity was evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. The HFE inhibited IL-4 production (IC50=48.73±0.45 μg/mL), and nitric oxide production (IC50=67.95±9.80 µg/mL), with slight inhibition of TNF-α and IL-6. The main active compounds were rhinacanthin-C (1.65±0.05% w/w) and rhinacanthin-N (0.20±0.01% w/w), which showed the highest effects on IL-4 secretion, nitric oxide, and inflammatory cytokines. The HFE and its active compounds were nontoxic to human cell lines, with over 70% cell viability, except for rhinacanthin-C at 100 μg/mL. Overall, the HFE effectively inhibited IL-4 secretion in RBL-2H3 cells, reduced nitric oxide release, and slightly decreased TNF-α and IL-6 levels. Rhinacanthin-C and rhinacanthin-N significantly regulated IL-4, nitric oxide, TNF-α, and IL-6 production. These findings suggest that the HFE could help alleviate allergies and acute skin inflammation.
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