Protection of 4-hydroxybenzyl-chitooligomers against inflammatory responses in Chang liver cells

https://doi.org/10.1016/j.ijbiomac.2014.01.064Get rights and content

Highlights

  • HB-COS inhibits inflammatory responses in Chang liver cells.

  • Inhibitory effects are due to suppression of NF-κB and MAPKs signaling pathways.

  • HB-COS has protective effect against hepatic inflammation.

Abstract

The aim of this study was to investigate anti-inflammatory activity of 4-hydroxybenzyl-chitooligomers (HB-COS) in Chang liver cells stimulated by a cytokine mixture. It was revealed that HB-COS decreased the level of nitric oxide and prostaglandin E2 (PGE2) production by diminishing the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) without significant cytotoxicity. Moreover, HB-COS exerted inhibitory effects on the production of pro-inflammatory mediator (interleukin-6) in Chang liver cells. Notably, HB-COS exhibited anti-inflammatory activities via blocking degradation of inhibitory kappa B alpha (IκB-α), translocation of nuclear factor kappa B (NF-κB), and phosphorylation of mitogen-activated protein kinases (MAPKs) in a dose-dependent manner. Collectively, these findings indicated that HB-COS possessed potential anti-inflammatory effects in Chang liver cells, and could be a useful therapeutic agent for the treatment of hepatic inflammatory diseases.

Introduction

Alcohol is mostly metabolized in the liver and excessive alcohol drinking can lead to acute and chronic liver diseases including hepatitis, fatty liver, liver cirrhosis and liver cancer [1], [2]. The liver is a major organ for the metabolism, disposition and toxicity of ingested ethanol. Chronic alcohol abuse, a major health concern, causes liver and pancreatic diseases and is known to impair hepatic alcohol dehydrogenase, and cause myocardial infarction pancreatitis and disorders of the immune, endocrine and reproductive systems [3], [4].

Inflammation has been recognized as a major risk factor for various human diseases. Prostaglandin and nitric oxide (NO) biosynthesis are involved in inflammation, and isoforms of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) are responsible for the production of a great amount of these pro-inflammatory mediators [5]. Nuclear factor kappa B (NF-κB) signaling pathway is found to be the major signaling mechanism involved in inflammation. NF-κB activation mediates the expression of a number of rapid response genes involved in the inflammatory responses to injury. Furthermore, mitogen-activated protein kinases (MAPKs) signaling pathways are also involved in modulation of inflammatory responses. Therefore, it is necessary to identify a potential candidate which has the ability to decrease the inflammatory responses at the molecular level [6], [7], [8], [9], [10].

Chitosan, a partially deacetylated polymer of N-acetyl glucosamine, is prepared by alkaline deacetylation of chitin. Chitooligomers (COS), partially hydrolyzed products of chitosan, is of great interest in pharmaceutical and medicinal applications due to their non-cytotoxic and high water soluble properties [11]. COS and COS derivatives have been demonstrated to exhibit a broad spectrum of biological activities for human health including anti-inflammatory [12], [13], antiallergy [14], [15], [16], antioxidative [17], [18], [19], [20], antihypertensive [21], [22], anticancer [23], anticoagulant [24], antibacterial [25], immuno-stimulant [26] and adipogenesis inhibitory [27] activities. In this study, we were interested in examining the anti-inflammatory mechanisms of HB-COS on Chang liver cells.

Section snippets

Materials

Chang liver cells (human hepatocyte derived cell line) were obtained from American Type Culture Collection (Manassas, VA, USA). COS (molecular weight 1.0–3.0 kDa) was donated by Kitto Life Co. (Seoul, Republic of Korea). MTT reagent (3-(4,5-dimethyl-2-yl)-2,5-diphenyltetrazolium bromide), 4-hydroxybenzaldehyde, Griess reagent, agarose and fetal bovine serum (FBS) were products of Sigma Chemical Co. (St. Louis, MO, USA). Dulbecco's Modified Eagle Medium (DMEM) medium, penicillin/streptomycin and

Chemical characterization of 4-hydroxybenzyl-chitooligomers

The mechanism envisaged for the reaction of COS with 4-hydroxybenzaldehyde is shown in Fig. 1. Amino groups on COS reacted with 4-hydroxybenzaldehyde to form a Schiff base intermediate, which is converted into N-alkyl COS (HB-COS). The reduction of aldimine was performed using NaBH4 [30], [31], [32], [33].

The structures of COS and HB-COS were confirmed via FT-IR (Fig. 2A and B) and 1H NMR (Fig. 2C and D) data as follows. According to the FT-IR spectrum (Fig. 2A), the peaks of COS were assigned

Conclusion

In conclusion, this study has determined that the grafting of 4-hydroxybenzyl onto COS enhances the suppressive effect of COS against inflammatory responses in CM-activated Chang liver cells. The suppressive effects were shown due to down-regulating the production of inflammatory mediators via inhibition of NF-κB activation and MAPKs phosphorylation. Our results indicate that HB-COS can contribute to suppression of inflammatory responses in live cells and might be a potential candidate for

Acknowledgement

This research is funded by Vietnam National Foundation for Science and Technology Development (NAFOSTED) under grant number 106.05-2011.36.

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