Anti-inflammatory activity of arctigenin from Forsythiae Fructus

doi:10.1016/j.jep.2007.11.030

Journal of Ethnopharmacology

Volume 116, Issue 2, 5 March 2008, Pages 305-312

https://doi.org/10.1016/j.jep.2007.11.030 Get rights and content

Abstract

Oleaceae Forsythiae Fructus has been used for anti-inflammatory, diuretics, antidote, and antibacterials in traditional herbal medicine. Our previous screening of medicinal plants showed that methanol (MeOH) extract of Forsythiae Fructus had significant anti-inflammatory activity, but the active ingredients remain unclear. For isolation of active ingredient of MeOH extract of Forsythiae Fructus, it was partitioned with n-hexane and ethylacetate (EtOAc), and arctigenin was isolated from EtOAc fraction by column chromatography with anti-inflammatory activity-guided separation. Its activity was evaluated in the animal models of inflammation including myeloperoxidase (MPO) and eosinophil peroxidase (EPO) activities in the edematous tissues homogenate, and silica-induced reactive oxygen species (ROS) production in the RAW 264.7 cell line. It was shown that arctigenin (100 mg/kg) had significantly decreased not only carrageenan-induced paw edema 3 and 4 h after injection of carrageenan, arachidonic acid (AA)-induced ear edema at a painting dose of 0.1–1.0 mg/ear, and acetic acid-induced writhing response and acetic acid-induced capillary permeability accentuation at an oral dose of 25–100, and 100 mg/kg, respectively, but also MPO and EPO activities at a painting dose of 0.1–1.0 mg/ear in the AA-induced edematous tissues homogenate as indicators of neutrophils and eosinophils recruitment into the inflamed tissue. Further, arctigenin (0.1–10 μM) also significantly inhibited the intracellular ROS production by silica. These results indicate that arctigenin is a bioactive agent of Forsythiae Fructus having significant anti-inflammatory action by inhibition of the exudation, and leukocytes recruitment into the inflamed tissues. The pharmacologic mechanism of action of arctigenin may be due to the inhibition of release/production of inflammatory mediators such as AA metabolites and free radicals.

Introduction

Oleaceae Forsythiae Fructus (known in Korea as Yeon-gyo) containing triterpenoids, lignans and its glycosides (Nishibe et al., 1977, Chiba et al., 1978, Chiba et al., 1979), is a traditional herbal medicine used as an anti-inflammatory, diuretics, antidotes, and antibacterials in Korea, China, and Japan (Lee et al., 1996). Recently, the methanol (MeOH) extract obtained from the dried fruit of Forsythia suspensa and its n-hexane fraction showed an anti-inflammatory effect on acetic acid-induced exudation and the active ingredient, identified as 3β-acetoxy-20,25-epoxydammarane-24-ol, a triterpenoid, was isolated from the n-hexane fraction by silica gel column chromatography (Ozaki et al., 2000). Kim et al. (2000) demonstrated that the butanol fraction of the aqueous extract of Forsythia koreana fruits also inhibits nitric oxide (NO) production and inducible nitric oxide synthesis (iNOS) gene expression in macrophage-like RAW 264.7 cells stimulated with interferon-γ plus lipopolysaccharide. The MeOH extract of Forsythiae Fructus was shown to have significant biologic activities on inflammatory processes and hypersensitivity (Kim et al., 1999), but its mechanism of action and the active ingredients of Forsythiae Fructus on inflammatory responses remain unclear.

In this study, MeOH extract of Forsythiae Fructus partitioned with n-hexane, ethyl acetate (EtOAc), chloroform (CHCl₃) and n-butanol (n-BuOH), and arctigenin was chromatographically isolated from Forsythiae Fructus by anti-inflammatory activity-guided separation. Experiments were carried out to evaluate the anti-inflammatory effects of arctigenin on experimental animal models of acute inflammation; carrageenan-induced paw edema in rats, arachidonic acid (AA)-induced ear edema in mice including myeloperoxidase (MPO) and eosinophil peroxidase (EPO) activities in the edematous tissues homogenate, acetic acid-induced writhing syndrome and acetic acid-induced capillary permeability accentuation in mice. Further, the intracellular reactive oxygen species (ROS) production was determined in the RAW 264.7 cell line.

Section snippets

Plant materials and chemicals

Forsythiae Fructus (3 kg), the fruits of Forsythia koreana, was purchased from Dong-Sung Pharmaceutical Co. Ltd. (Gu Ri City, Kyung Ki-Do, Korea), and identified by Professor W.K. Whang (Division of Natural Products, College of Pharmacy, Chung-Ang University, Korea). A voucher specimen was deposited in College of Pharmacy, Chung-Ang University. Dulbecco's modified Eagle minimum essential medium (DMEM), fetal bovine serum (FBS), and an antibiotic–antimycotic mixture were purchased from Invitrogen

Inhibition of AA-induced ear edema

The painting of AA (2 mg/ear) significantly induced ear edema 40 min after application of AA compared to the vehicle controls in mice (60.9 ± 6.9%, P < 0.01). The MeOH extract of Forsythiae Fructus and all of its fractions at a painting dose of 0.1–1.0 mg/ear inhibited significantly the AA-induced ear edema in mice compared with controls on the dose-dependent manner, respectively. Anti-inflammatory activities of H₂O (32.5 ± 4.9%, P < 0.01), EtOAc (30.9 ± 4.1%, P < 0.01) and BuOH fractions (29.6 ± 3.6%, P < 0.01)

Discussion and conclusions

In this experiments, it was shown that arctigenin have significant anti-inflammatory action. Arctigenin indicates a representative dibenzylbutyrolactone lignan with anti-tumor, anti-viral, and Ca²⁺ antagonist activities (Kim et al., 2006), and a suppressive agents at the level of the activation of heat shock transcription factor, the induction of mRNA, and the synthesis and accumulation of heat shock proteins, which are involved in protecting cells and in the pathophysiology of diseases such as

Acknowledgement

This Research was supported by the Chung-Ang University Research Grants in 2007.

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^☆: This present study was performed according to international, national and institutional rules considering animal experiments, clinical studies and biodiversity rights, and have ethnopharmacological importance for evaluation of anti-inflammatory pharmacological actions and its action mechanism of medicinal plant, Forsythiae Fructus.

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Anti-inflammatory activity of arctigenin from Forsythiae Fructus☆

Abstract

Introduction

Section snippets

Plant materials and chemicals

Inhibition of AA-induced ear edema

Discussion and conclusions

Acknowledgement

Journal of Investigative Dermatology

European Journal of Pharmacology

Journal of Biological Chemistry

Journal of Ethnopharmacology

Journal of Ethnopharmacology

Phytochemistry

Free Radical Biology and Medicine

Journal of Immunological Methods

Journal of Biological Chemistry

Studies on the antinociceptive action of α-agonist drugs and their interactions with opioid mechanisms

British Journal of Pharmacology

Modulation of mouse ear edema by cyclooxygenase and lipoxygenase inhibitors and other pharmacologic agents

Agents Actions

Studies on the chinese crude drug “Forsythiae Fructus” (III). On the constituents of fruits of Forsythia viridissima and F. suspensa

Shoyakugaku Zasshi

Studies on the chinese crude drug “Forsythiae Fructus” (IV). On the constituents of fruits of Forsythia koreana and Forsythiae Fructus from Korea on the market

Shoyakugaku Zasshi

Inhibition of arachidonic acid-induced ear edema as a model for assessing topical anti-inflammatory compounds

Agents Actions

(−)-Arctigenin as a lead structure for inhibitors of human immunodeficiency virus type-1 integrase

Journal of Medicinal Chemistry

Reactive oxygen intermediates as regulators of TNF-α production in rat lung inflammation induced by silica

Journal of Immunology

Role of endogenous catecholamines in the anti-inflammatory activity of α-adrenoceptor blocking agents

British Journal of Pharmacology

Arctigenin from Fructus Arctii is a novel suppressor of heat shock response in mammalian cells

Cell Stress Chaperones