Effects of polysaccharides from abalone (Haliotis discus hannai Ino) on HepG2 cell proliferation

https://doi.org/10.1016/j.ijbiomac.2014.01.032Get rights and content

Abstract

Three polysaccharides, AAP, AVAP I, and AVAP II, were isolated from abalone Haliotis discus hannai Ino. The polysaccharides’ compositions were analysed, and their effects on HepG2 cell proliferation were assessed. AVAP I had a greater growth-stimulatory effect than AAP or AVAP II. The oligosaccharide of AVAP I (Oli-AVAP I) exhibited the same growth effects, but rhamnose, the primary monosaccharide of AVAP I and Oli-AVAP I, did not exhibit this activity. Moreover, AVAP I dramatically reduced the mRNA levels of CDK6 and Cyclin E1 but significantly increased Cyclin B1, CDK1 and Cyclin F. Interestingly, AVAP I remained able to induce cell proliferation in a low serum concentration medium. AVAP I could therefore promote HepG2 cell proliferation by regulating gene expression and accelerating the cell cycle process. AVAP I may be useful as a serum supplement for stimulating the proliferation of mammalian cells. Our results offer a comprehensive method for utilising the abalone viscera, which is usually discarded as waste.

Introduction

Abalone, Haliotis discus hannai Ino, is a large, single-shelled marine mollusc of the genus Haliotis. Abalone is widely cultured in East Asia as an economically important food resource [1]. Owing to its nutritive and pharmaceutical value, abalone has received extensive attention [1], [2], [3], [4], [5]. Abalone viscera accounts for 15–25% of the total body weight of the abalone, and this visceral matter is normally discarded as industrial waste [3]. The visceral waste contains many proteins, polysaccharides and fatty acids [4]. A substantial number of studies have been published regarding the activities of abalone polysaccharides, including antitumour [4], immunoregulatory [5] and antioxidative [6] activities. However, there are few reports concerning the cytoproliferative activity of the polysaccharides from pleopods and from the viscera of H. discus hannai Ino.

There has been a rapid development in the fields of cell therapy and regenerative medicine and in the production of bio-medicine using mammalian cell culture [7]. Accelerating cell proliferation is fundamental for these applications. It has been reported that sericin, a protein derived from silkworms, is a good supplement for cell culture media to accelerate the proliferation of mammalian cells [8], [9]. Sericin was added to freezing media as an alternative to foetal bovine serum (FBS) and successfully improved the survival of various cell lines during cryopreservation [10], [11]. Cell proliferation depends on intracellular signal transduction mediated by receptors, such as enzyme-linked receptors and protein degradation-dependent receptors [12]. Cyclins are positive regulators of cell cycle progression that are produced at specific periods during the cell cycle, and their expression levels and locations are tightly controlled [13]. Cyclins are the positive regulatory subunits of a class of protein kinases termed cyclin-dependent kinases (CDKs) [14]. Earlier studies have reported that sulphated polysaccharides from the sea cucumber Stichopus iaponicus could induce the proliferation of rat astrocytes by causing the accumulation of Cyclin D1, which integrates with the extracellular signals to activate CDK4 and/or CDK6 [15]. Progesterone was also able to promote the viability of mouse embryonic stem cells through the up-regulation of cyclins and cyclin-dependent kinases [16].

In the present study, three types of polysaccharides, AAP, AVAP I, and AVAP II, were isolated from the abalone H. discus hannai Ino and purified. The polysaccharides’ compositions were subsequently identified. We investigated the cell proliferation activity of these polysaccharides, and further studies examined AVAP I and the relationship between its composition and its effects on cell proliferation. We also examined whether the cell cycle-regulating genes were involved in inducing proliferation. Our results indicated that AVAP I had a significant effect on HepG2 cell proliferation and could be used as a supplement for stimulating the proliferation of mammalian cells.

Section snippets

Materials

Fresh abalones, cultivated in Jiaonan, Shandong Province, China, were collected in the Nanshan aquatic market of Qingdao, shelled, eviscerated, vacuum freeze-dried and crushed. DEAE-cellulose anion-exchange resin was from Whatman (Brentford, England), and Sephacryl™ S-300 HR was from Pharmacia Co. (Uppsala, Sweden). A TSK G4000 PWXL column was from TOSOH Biosep (Tokyo, Japan). The human hepatoma cell line HepG2 was purchased from the Institute of Basic Medicine, Shandong Academy of Medical

Isolation, purification and chemical analysis of the polysaccharides

Three polysaccharides, AAP, AVAP I and AVAP II, were isolated and purified from the pleopods and viscera of H. discus hannai Ino. The extracted crude polysaccharide was firstly purified by ion-exchange chromatography on a DEAE-cellulose column and the main polysaccharide fraction from the pleopods was eluted with 0.42–0.60 mol/L NaCl and collected, two fractions from the viscera were respectively eluted with 0.28–0.40 mol/L NaCl and 0.44–0.56 mol/L NaCl and collected. The pooled polysaccharide

Discussion

In the present study, we isolated three polysaccharides (AAP, AVAP I and AVAP II) from the abalone H. discus hannai Ino. The composition analysis of the polysaccharides revealed that all three were sulphated, and their sulphate contents were 21.5%, 15.2% and 20.9% for AAP, AVAP I and AVAP II, respectively. The monosaccharide composition of AVAP I and AVAP II was similar and primarily composed of Rha, GlcA and Gal, but the composition of AAP was different, containing GlcA, GalN, Fuc and Gal. Wu

Acknowledgments

This study was supported by the National Marine Public Welfare Scientific Research Project of China (No. 201105029) and the Programme for Changjiang Scholars and Innovative Research Team in University (IRT1188).

References (34)

  • B.W. Zhu et al.

    Food Chemistry

    (2011)
  • C. Yoneda et al.

    Comparative Biochemistry and Physiology Part B

    (2000)
  • A. Ogawa et al.

    Journal of Bioscience and Bioengineering

    (2004)
  • X.H. Sheng et al.

    Neuroscience Letters

    (2011)
  • G.Y. Li et al.

    International Journal of Biological Macromolecules

    (2011)
  • O.H. Lowry et al.

    Journal of Biological Chemistry

    (1951)
  • S.I. Ohira et al.

    Journal of Chromatography A

    (2006)
  • D.J. Strydom

    Journal of Chromatography A

    (1994)
  • Z.Q. Huang et al.

    Biochemical and Biophysical Research Communications

    (2012)
  • Y.J. Zhang et al.

    Journal of Bioscience and Bioengineering

    (2010)
  • M. Kunou et al.

    Carbohydrate Polymers

    (1995)
  • V. Ravelojaona et al.

    Pathologie Biologie

    (2008)
  • G. Faury et al.

    Biochimica et Biophysica Acta

    (2008)
  • E. Andres et al.

    Pathologie Biologie

    (2006)
  • Y. Xiong et al.

    Cell

    (1992)
  • D.Y. Zhou et al.

    Journal of Food Processing and Preservation

    (2012)
  • B.W. Zhu et al.

    European Food Research and Technology

    (2008)
  • Cited by (32)

    • New polysaccharides extracted from Malcolmia triloba: Structure characterization, biological properties and application to beef meat preservation

      2022, Journal of Food Composition and Analysis
      Citation Excerpt :

      The weak peak detected at about 2929 cm−1 was attributed to the CH stretching vibration of free sugars (Zhu et al., 2013). The peak at 1644 cm−1 was attributed to the bond stretching of carboxylate correspond to the presence of proteins (Wang et al., 2014a, 2014b). The peak around 1628 and 1636 cm−1 assigned to the bound water, While the peak at 1401 cm-1 was attributed to COOH group indicating uronic acids (Jia et al., 2013).

    • Effect of intake pattern of sulfated polysaccharides on its biological activity in high fat diet-fed mice

      2019, International Journal of Biological Macromolecules
      Citation Excerpt :

      Recent study indicated that the main structure residue of AGSP was determined as →3)-GlcA (1 → 3)-Gal (1 → with sulfated branches comprised of Gal and Glc, and →4)-β-GlcA (1 → 2)-α-Man (1 → residue was also found [15]. Previous studies showed that AGSP could play many health-promoting roles in the human health, e.g. immunomodulatory, anticoagulant, and anti-tumor activities [16–18]. In this study, an obese mice model was established by feeding mice with a HFD, and the effect of intake pattern of AGSP on its bioactivity was evaluated by analyzing physiological parameters and the gut microbiota and its metabolites.

    • Sulfated polysaccharide isolated from Globularia alypum L.: Structural characterization, in vivo and in vitro anticoagulant activity, and toxicological profile

      2019, International Journal of Biological Macromolecules
      Citation Excerpt :

      As shown in Fig. 2, the band at 3271 cm−1 is assigned to the hydrogen bonded OH stretching vibration; a weak band observed at around 3008 cm−1 is assigned to a CH stretching vibration [32]. Furthermore, the band at 1743 cm−1 was attributed to the bond stretching vibrations of (CO) bonds of the carboxylate group [33]. The most important peaks were those observed at 1541 cm−1 deriving from the bending vibration of the stretching vibration of the ester sulfate groups (SO) as were previously described in the literature [34].

    View all citing articles on Scopus
    1

    These authors have the same attribute to the article.

    View full text