Bioactive hyaluronic acid fragments inhibit lipopolysaccharide- induced inflammatory responses via the Toll-like receptor 4 signaling pathway

Abstract

The high- and the low-molecular weight hyaluronic acids (HMW-HA and LMW-HA, respectively) showed different biological activities in inflammation. However, the role of LMW-HA in inflammatory response is controversial. In this study, we aimed to investigate the effect of bioactive hyaluronan (B-HA) on lipopolysaccharide (LPS)-induced inflammatory responses in human macrophages and mice. B-HA was produced from HA treated with glycosylated recombinant human hyaluronidase PH20. Human THP-1 cells were induced to differentiate into macrophages. THP-1-derived macrophages were treated with B-HA, LPS, or B-HA+LPS. The mRNA expression and the production of inflammatory cytokines were determined using quantitative real-time PCR and enzyme-linked immunosorbent assay. The phosphorylation levels of proteins in the nuclear factor-kB (NF-kB), mitogen-activated protein kinase (MAPK), and IRF-3 signaling pathways were measured using Western blot. The in vivo efficacy of B-HA was assessed in a mouse model of LPS-induced inflammation. Results showed that B-HA inhibited the expression of TNF-α, IL-6, IL-1, and IFN-β, and enhanced the expression of the anti-inflammatory cytokine IL-10 in LPS-induced inflammatory responses in THP-1-derived macrophages and in vivo. B-HA significantly suppressed the phosphorylation of the TLR4 signaling pathway proteins p65, IKKα/β, IkBα, JNK1/2, ERK1/2, p38, and IRF-3. In conclusion, our results demonstrated that the B-HA attenuated the LPS-stimulated inflammatory response by inhibiting the activation of the TLR4 signaling pathway. B-HA could be a potential anti-inflammatory drug in the treatment of inflammatory disease.

Keywords

bioactive hyaluronan / lipopolysaccharide / inflammatory cytokines / TLR4 / human macrophages

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Acknowledgements

We would like to thank Editage for English language editing and Publication Support. This work was supported by grants from the Chinese National Twelfth Five-year Plan Project (No. 2013ZX09J-13110-05B, MW) and the Social Development Project of Jiangsu Province (No. BE2015651, JZ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Compliance with ethics guidelines

Na You, Sasa Chu, Binggang Cai, Youfang Gao, Mizhou Hui, Jin Zhu, and Maorong Wang declare that they have no conflict of interest. All institutional and national guidelines for the care and use of laboratory animals were followed.

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