Anti-Inflammatory Effects of Methanol Extract, Hexane, Ethyl Acetate, and Butanol Fraction of Piper crocatum Ruiz & Pav Leaves on Lipopolysaccharide-induced RAW 264.7 Cells

Abstract

Piper crocatum Ruiz & Pav is a traditional Indonesian plant that is empirically used to treat various diseases. Several studies have stated that Piper crocatum leaves extract has anti-inflammatory effects. Piper crocatum leaves contain different secondary metabolites alkaloids, flavonoids, tannin-polyphenols, steroids, terpenoids, and saponins. The purpose of this study was to determine the anti-inflammatory effect of Piper crocatum leaves extract and fraction on the production of nitric oxide in lipopolysaccharideinduced RAW 264.7 cells. Anti-inflammatory effect of methanol extract of Piper crocatum leaves (MEPC), n-hexane fraction of Piper crocatum leaves (HFPC), ethyl acetate fraction of Piper crocatum leaves (EAFPC), and n-butanol fraction of Piper crocatum leaves (BFPC) against RAW 264 cell macrophages, 7 Lipopolysaccharide-induced (LPS). Extraction of Piper crocatum leaves was the maceration method. Cell viability was determined by the MTT method in the concentration range of 12.5;25;50;100;200 ug/ mL. The anti-inflammatory effects of MEPC, HFPC, EAFPC and BFPC were tested against nitric oxide (NO) production inhibition in LPS-induced RAW 264.7 macrophage cells. NO levels were determined by the NO Colorimetric Assay Kit as measured by the ELISA plate reader. The viability test of MEPC, HFPC, EAFPC, and BFPC from Piper crocatum leaves showed the concentrations that gave the percentage of viability above 80% were concentrations of 100, 50, and 25 g/mL. MEPC, HFPC, EAFPC, and BFPC at concentrations of 100, 50, 25 μg/mL could significantly inhibit NO production with p<0.05 (p=0.000). The highest NO production inhibition effect was on EAFPC. MEPC, HFPC, EAFPC, and BFPC have antiinflammatory effects, with the highest effect on EAFPC. Key words: Piper crocatum, Anti-inflammatory effect, LPS, Nitric oxide.