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

Nilda Lely,Fatma Sri Wahyuni,Yufri Aldi,Helmi Arifin

doi:10.5530/pj.2021.13.169

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.

Highlights

Inflammation is a physiological response to stimuli such as infection and tissue injury
NO is the main product controlled by nitric oxide synthase (NOS), such as iNOS. iNOS is induced in macrophages so that its activation leads to organ destruction in several inflammatory and autoimmune diseases. 2,3
Cell viability test of RAW 264.7 on the administration of methanol extract of Piper crocatum leaves (MEPC), ethyl acetate fraction of Piper crocatum leaves (EAFPC), hexane fraction of Piper crocatum leaves (HFPC) and butanol fraction of Piper crocatum leaves (BFPC) from red betel leaf showed a decrease in the number of live cells with increasing concentration

Summary

Introduction

Inflammation is a physiological response to stimuli such as infection and tissue injury. This stimulation causes the release of inflammatory mediators such as nitric oxide, histamine, serotonin, bradykinin, and prostaglandins which cause an inflammatory reaction in the form of heat, pain, redness, swelling, and accompanied by impaired function. 1. Lipopolysaccharide is a component of the cell wall of gram-negative bacteria, the polyclonal activator of the immune system, stimulates the release of various inflammatory mediators, including nitric oxide (NO), prostaglandin E2, proinflammatory cytokines such as IL-1, IL-6, IL-12, IL-18, TNF-α and TNF-β. Activated macrophages are the main effector cells in the host defense against bacteria through the production of nitric oxide (NO), which is cytotoxic to parasites. Red betel is often used to treat diabetes mellitus, hepatitis, kidney stones, lowering cholesterol, preventing stroke, gout, hypertension, coronary heart disease, uterine cancer, breast cancer, hemorrhoids, tuberculosis, toothache medicine, canker sores, body odor. , venereal disease, inflammation of the liver, inflammation of the prostate, inflammation of the eye, vaginal discharge, ulcers, fatigue, joint pain, smoothing the skin, inflammation of the ear, cough medicine, inflammation of the lungs, inflammation of the throat, inflammation of the gums, inflammation of the breast, nose bleed, and coughing up blood.[4]

Objectives

Methods

Results

Conclusion