Elsevier

Phytomedicine

Volume 22, Issue 2, 15 February 2015, Pages 326-332
Phytomedicine

Traditional herbal formula Jakyakgamcho-tang (Paeonia lactiflora and Glycyrrhiza uralensis) impairs inflammatory chemokine production by inhibiting activation of STAT1 and NF-κB in HaCaT cells

https://doi.org/10.1016/j.phymed.2014.12.002Get rights and content

Abstract

A traditional herbal formula Jakyakgamcho-tang (JYGCT; Paeonia lactiflora and Glycyrrhiza uralensis) has been used for treatment of backache, muscle pain, acute abdominal pain, neuralgia, bronchial asthma, and painful peripheral neuropathy in Oriental medicine. We report on our experiments using the HaCaT human keratinocyte cell line showing that a traditional herbal formula JYGCT has inhibitory effects on inflammatory responses in skin.

Stimulation with tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) caused a significant increase in the production of the following chemokines: thymus- and activation-regulated chemokine (TARC)/CCL17; macrophage-derived chemokine (MDC)/CCL22; regulated on activation, normal T-cell expressed and secreted (RANTES)/CCL5; and interleukin-8 (IL-8) in HaCaT cells. By contrast, treatment with JYGCT extract significantly reduced the production of TARC, MDC, RANTES, and IL-8, but caused no cytotoxicity, compared with TNF-α and IFN-γ-treated control cells. Consistently, JYGCT extract downregulated the mRNA expression of TARC, MDC, RANTES, and IL-8 induced by TNF-α and IFN-γ in a dose-dependent manner. In addition, TNF-α and IFN-γ markedly increased the phosphorylation of signal transducer and activator of transcription 1 (STAT1) and the nuclear translocation of nuclear factor kappa B (NF-κB) in HaCaT cells. By contrast, TNF-α and IFN-γ-induced activation of STAT1 and NF-κB activation was inhibited by JYGCT treatment in a dose-dependent manner.

Our data indicate that JYGCT attenuates TNF-α and IFN-γ-mediated chemokine production by targeting the STAT1 and NF-κB signalling in keratinocytes. Our findings suggest that JYGCT has potential as a therapeutic drug candidate for the treatment of inflammatory skin diseases.

Introduction

Skin immune responses are important aspects of the host defense mechanisms against microorganisms (Pasparakis et al. 2014). Disruption of skin immunity can cause chronic inflammatory skin diseases. T-helper 2 type (Th2) cells, including monocytes, macrophages, eosinophils, and mast cells are crucial cells associated with the development of skin diseases (Meagher et al., 2002, Werfel, 2009). Th2 cells are attracted to the skin by inflammatory chemokines, such as regulated on activation, normal T-cell expressed and secreted (RANTES); thymus and activation regulated chemokine (TARC); and macrophage-derived chemokine (MDC). These chemokines interact with the cells via extracellular receptor proteins (Kaburagi et al., 2001, Kakinuma et al., 2002, Shimada et al., 2004). Epidermal keratinocytes play an important role in the control of skin inflammation through their production of inflammatory chemokines (Albanesi 2010). Therefore, chemokines are considered as pivotal mediators in the development of inflammatory skin diseases. Several studies have reported that transcription factors such as signal transducer and activator of transcription 1 (STAT1) and nuclear factor kappa B (NF-κB) play critical roles in the chemokine production mediated by tumour necrosis factor-alpha (TNF-α) and/or interferon-gamma (IFN-γ) in keratinocytes (Han et al., 2011, Ju et al., 2009, Kwon et al., 2012).

Accumulating evidence shows that many traditional herbal medicines have anti-inflammatory activities in the skin (Hon et al., 2011, Hughes et al., 2007, Lee do et al., 2009a, Lee do et al., 2009b). Jakyakgamcho-tang (JYGCT), also called Shaoyao-gancao-tang in Chinese and Shakuyaku-kanzo-to in Japanese, is a traditional herbal medicine comprising two medicinal herbs, Paeonia lactiflora Pall. (Paeoniae Radix) and Glycyrrhiza uralensis Fisch. (Glycyrrhizae Radix et Rhizoma) in a 2:1 ratio (Kim 2010). JYGCT is used clinically and pharmacologically in the treatment of backache, muscle pain, acute abdominal pain, neuralgia, bronchial asthma, and painful peripheral neuropathy (Hidaka et al., 2009, Hinoshita et al., 2003, Kim, 2010, Tsuji et al., 2012, Wu et al., 2007). However, an inhibitory effect of JYGCT on skin inflammation has not been reported. We here report on our experiments in which we stimulated HaCaT human keratinocytes with TNF-α and IFN-γ to investigate the anti-inflammatory activities of JYGCT.

Section snippets

Chemicals and reagents

Gallic acid, (+)-catechin, and benzoic acid were purchased from Sigma-Aldrich (St. Louis, MO). Oxypaeoniflorin, isoliquiritin, ononin, and benzoylpaeoniflorin were obtained from Biopurify Phytochemicals (Chengdu, China). Albiflorin, paeoniflorin, and glycyrrhizin were purchased from Wako (Osaka, Japan). Liquiritin was supplied by NPC Biotechnology (Geumsan, Korea). The purity of all reference compounds was ≥98.0%. High performance liquid chromatography (HPLC)-grade reagents, methanol,

Quantitative analysis of 11 marker components in JYGCT (Paeonia lactiflora and Glycyrrhiza uralensis)

The HPLC chromatogram was obtained using two mobile phase systems (0.1% formic acid in water and acetonitrile) with a gradient elution. Under optimized chromatography conditions, all components were separated before 45 min and showed resolution above 1.97 in sample analysis. A typical HPLC chromatogram of JYGCT water extract is shown in Fig. 1. The calibration curves of all compounds showing good linearity with r2 ≥ 0.9998 in seven different concentration ranges and other parameters are shown

Discussion

Herbal medicines are attractive drug candidates for providing a new therapeutic approach and are attracting increasing attention as treatment for inflammatory skin diseases (Ko et al. 2006). In the present study, we investigated the in vitro anti-inflammatory effects of a traditional herbal formula JYGCT in the skin using a human keratinocyte cell line. Quantitative analysis of marker components in JYGCT was performed using HPLC–PDA. The main constituents of two herbal medicines P. lactiflora

Conflicts of interest

The authors have no conflicts of interest to declare.

Acknowledgement

This work was supported by a Grant from the Korea Institute of Oriental Medicine (no. K14030).

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