Immunomodulatory effects of hydroxyethylated Hericium erinaceus polysaccharide on macrophages RAW264.7

doi:10.1016/j.ijbiomac.2017.07.104

International Journal of Biological Macromolecules

Volume 105, Part 1, December 2017, Pages 879-885

https://doi.org/10.1016/j.ijbiomac.2017.07.104 Get rights and content

Abstract

Hericium erinaceus polysaccharide (HEP) has been shown to possess a variety of biological activities. In present study, HEP was successfully modified to obtain its hydroxyethylated derivative hHEP. Its potential immunomodulatory activities on RAW264.7 macrophages were investigated. Results showed that the hHEP were significantly stronger than that of the corresponding unmodified polysaccharide, HEP. Meanwhile, the NO, IL-6 and TNF-α production activities of macrophages were enhanced in the RAW264.7 macrophages by stimulation of hHEP. In addition, the hHEP increase significantly higher iNOS expression than HEP. These results indicated that the hydroxyethylated derivative hHEP could enhance the activation of peritoneal macrophages, and hydroxyethylation modification can enhance the immunomodulation function of HEP.

Introduction

Hericium erinaceus (Bull.:Fr.) Pers. has been used as functional foods and herbal folk medicine in China for a long time [1], [2], [3]. It belongs to the family Hericiaceae. It is a traditional medicinal food for the prevention and treatment of chronic atrophic gastritis (CAG), duodenal ulcers and several other diseases [2]. Several classes of phytochemicals, such as polysaccharides, ketones, terpenoids and phenolic compounds have been isolated and identified from the H. erinaceus. [4], [5], [6], [7]. The polysaccharides derived from H. erinaceus is a main active ingredient, and has various biological activities such as anti-tumor, anti-oxidation, antimicrobial, anti-inflammatory and immunomodulatory [8], [9], [10], [11], [12].

Polysaccharides is a kind of important biological macromolecules, but their bioactivities are not always in the condition of satisfaction [13], [14]. Therefore, molecular modification of polysaccharide is an significant means to enhance its biological activities. After modification, the inactive or less active polysaccharide is greatly improved. As an effective chemical modification method of polysaccharide, hydroxyethylation is interesting due to operate simple and its significant effects on improve the physiological properties [15]. Many researchers have demonstrated that hydroxyethylated modification polysaccharides were significantly improved their immune activity [16], [17]. However, the hydroxyethylated modification influence on the immunoregulatory effects of polysaccharides from H. erinaceus remains unclear.

Macrophages is one kind of important immune cells and the body's first line of defense against infection. It play a critical role in the innate and adaptive immune responses [18]. Under normal conditions, macrophages are in the static state. But when the body is stimulated by pathologic stimuli or injury, macrophages could kill these foreign matters indirectly via the secretion of macrophage-derived biological factors, including nitric oxide (NO), iNOS, reactive oxygen species (ROS), and several cytokines such as IL-6, IL-8, TNF-α and so on [19], [20]. Thus, macrophages have been broadly used to evaluate the immunomodulatory potential of bioactive polysaccharides through multiple parameters analysis.

Based on the previous studies [21], we prepared the hydroxyethylation derivative of H. erinaceus polysaccharide and its immunomodulatory activities were evaluated and compared. The immunomodulatory effects of hydroxyethylated modification polysaccharides from H. erinaceus on RAW264.7 macrophages were determined by assessing NO, IL-6, TNF-α production, iNOS expression and phagocytosis ability. The aim is to obtain a good immune activity of the hydroxyethylation polysaccharide from H. erinaceus, and provide theoretical basis for further clinical application of immunologic adjuvant and immune enhancer.

Section snippets

Materials and chemicals

H. erinaceus was obtained from Huishihao Pharmacy in Fujian Province of China. The identities of the plants were verified by Dr. Yifan Huang and deposited (voucher no. 20160715) in Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health in Fujian province, Fujian Agriculture and Forestry University.

RAW264.7 cells were obtained from institute of cell biology, Chinese academy sciences (Shanghai, China). DMEM was purchased from Gibco (Invitrogen Corporation, U.S.).

The infrared spectra of HEP and hHEP

The FT-IR spectra of HEP and hHEP were illustrated in Fig. 1. In the spectra of HEP and hHEP, there were three band, one appeared in 3600–3200 cm⁻¹ corresponding to the hydroxyl stretching vibration, the second appeared in the 2922 cm⁻¹ or 2924 cm⁻¹ corresponding to methyl C single bond H stretching vibration, the third appeared in 1400–1000 cm⁻¹ corresponding to COC stretching vibration. This indicated that the HEP and hHEP were polysaccharides.

As compared with the spectrum of HEP, the spectrum of hHEP

Discussion

Macrophages are important immune cells with diverse functions. Macrophages could phagocytose and kill pathogenic microorganisms, release some inflammatory mediators and active cytokines to defense against invading pathogens [25], [26]. It could also present the pathogen to the lymphatic tissue, which activating the specific immune response and regulating the body immunity [27]. Therefore, in the present study, RAW264.7 macrophages were used as an in vitro cell model to investigate the immune

Conclusion

In this study, a homogeneous H. erinaceus polysaccharides were modified to obtain its hydroxyethylation derivatives, hHEP. It is the first report on the evaluation of the composition and immunomodulatory effects of hydroxyethylated polysaccharides from H. erinaceus. We have shown that hHEP promoted the phagocytic function. Moreover, hHEP showed stronger immunomodulatory activities than HEP in term of stimulating the production of NO and cytokines. Finally, hHEP enhanced the iNOS expression. The

Conflict of interests

Authors declare that there is no conflict of interests.

Acknowledgements

The project was supported by National Natural Science Foundation of China (NSFC31402214), Natural Science Foundation of Fujian Province of China (2016J01697), The Education Department Foundation of Fujian Province (JAT160139). We are grateful to all other staff in the Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health in Fujian province for their assistance in the experiments.

References (44)

J.H. Liu et al.
Anti-Helicobacter pylori activity of bioactive components isolated from Hericium erinaceus
J. Ethnopharmacol.
(2016)
D. Rajagopalu et al.
Recovery of laccase from processed Hericium erinaceus (Bull.:Fr) Pers. fruiting bodies in aqueous two-phase system
J. Biosci. Bioeng.
(2016)
Q.Z. Li et al.
Structure elucidation of a bioactive polysaccharide from fruiting bodies of Hericium erinaceus in different maturation stages
Carbohydr. Polym.
(2016)
S.R. Lee et al.
A new cerebroside from the fruiting bodies of Hericium erinaceus and its applicability to cancer treatment
Bioorg. Med. Chem. Lett.
(2015)
C.C. Zhang et al.
Chemical constituents from Hericium erinaceus and their ability to stimulate NGF-mediated neurite outgrowth on PC12 cells
Bioorg. Med. Chem. Lett.
(2015)
H. Lakshmanan et al.
Haematological, biochemical and histopathological aspects of Hericium erinaceus ingestion in a rodent model: a sub-chronic toxicological assessment
J. Ethnopharmacol.
(2016)
G. Li et al.
Anticancer potential of Hericium erinaceus extracts against human gastrointestinal cancers
J. Ethnopharmacol.
(2014)
X.R. He et al.
Structures, biological activities, and industrial applications of the polysaccharides from Hericium erinaceus (Lion's Mane) mushroom: a review
Int. J. Biol. Macromol.
(2017)
Z.H. Han et al.
Evaluation of in vivo antioxidant activity of Hericium erinaceus polysaccharides
Int. J. Biol. Macromol.
(2013)
Y. Zhu et al.
Preparation characterization, and anti-Helicobacter pylori activity of Bi3+-Hericium erinaceus polysaccharide complex
Carbohydr. Polym.
(2014)

S.C. Sheu et al.

Immunomodulatory effects of polysaccharides isolated from Hericium erinaceus on dendritic cells

Process Biochem.

(2013)

W. Li et al.

Sterols, aromatic compounds, and cerebrosides from the Hericium erinaceus fruiting body

Biochem. Syst. Ecol.

(2017)

U. Surayot et al.

Structural effects of sulfated polysaccharides from Codium fragile on NK cell activation and cytotoxicity

Int. J. Biol. Macromol.

(2017)

X. Liu et al.

Immunomodulatory effects of an acetylated Cyclocarya paliurus polysaccharide on murine macrophages RAW264.7

Int. J. Biol. Macromol.

(2017)

N.P. Alamuru-Yellapragada et al.

In-house made nucleofection buffer for efficient and cost effective transfection of RAW 264.7 macrophages

Biochem. Bioph. Res. Co.

(2017)

I.A. Schepetkin et al.

Botanical polysaccharides: macrophage immunomodulation and therapeutic potential

Int. Immunopharmacol.

(2006)

T. Qin et al.

Selenizing Hericium erinaceus polysaccharides induces dendritic cells maturation through MAPK and NF-κB signaling pathways

Int. J. Biol. Macromol.

(2017)

Z.Y. Wang et al.

Characterization and antioxidant activity in vitro and in vivo of polysaccharide purified from Rana chensinensis skin

Carbohydr. Polym.

(2015)

F.Y. Liu et al.

Immunomodulatory effects of xanthan gum in LPS-stimulated RAW 264.7 macrophages

Carbohydr. Polym.

(2017)

H. Sun et al.

Activation of RAW264.7 macrophages by the polysaccharide from the roots of Actinidia eriantha and its molecular mechanisms

Carbohydr. Polym.

(2015)

S. Gordon

Phagocytosis an immunobiologic process

Immunity

(2016)

P.M. Zheng et al.

Characterization of polysaccharides extracted from Platycodon grandiflorus (Jacq.) A.DC. affecting activation of chicken peritoneal macrophages

Int. J. Biol. Macromol.

(2017)

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¹: These authors equally contributed to this work.

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Immunomodulatory effects of hydroxyethylated Hericium erinaceus polysaccharide on macrophages RAW264.7

Abstract

Introduction

Section snippets

Materials and chemicals

The infrared spectra of HEP and hHEP

Discussion

Conclusion

Conflict of interests

Acknowledgements

J. Ethnopharmacol.

J. Biosci. Bioeng.

Carbohydr. Polym.

Bioorg. Med. Chem. Lett.

Bioorg. Med. Chem. Lett.

J. Ethnopharmacol.

J. Ethnopharmacol.

Int. J. Biol. Macromol.

Int. J. Biol. Macromol.

Carbohydr. Polym.

Process Biochem.

Biochem. Syst. Ecol.

Int. J. Biol. Macromol.

Int. J. Biol. Macromol.

Biochem. Bioph. Res. Co.

Int. Immunopharmacol.

Int. J. Biol. Macromol.

Carbohydr. Polym.

Carbohydr. Polym.

Carbohydr. Polym.

Immunity

Int. J. Biol. Macromol.