Characterization and Biological Activity of Taishan Pinus massoniana Pollen Polysaccharide In Vitro
March
Characterization and Biological Activity of Taishan Pinus massoniana Pollen Polysaccharide In Vitro
Academic Editor: Charles J. Russell 0 1
St. Jude Children's Research Hospital 0 1
UNITED STATES 0 1
Shifa Yang 0 1
Kai Wei 0 1
Fengjuan Jia 0 1
Xue Zhao 0 1
Guolin Cui 0 1
Fanxia Guo 0 1
Ruiliang Zhu 0 1
0 Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science and Technology, Shandong Agricultural University , Shandong, Taian, 271018 , PR China
1 Funding: This study was funded by Technology Development Plan of Shandong Province (2012GNC11020) and Specific Technology Development Projects of Taian (20103001)
Taishan Pinus massoniana pollen polysaccharide (TPPPS) improves cellular and humoral immune responses of animals and is a novel potential immunomodulator. However, the components of TPPPS have not been recognized. To investigate the composition of TPPPS, crude polysaccharide was obtained from Taishan P. massoniana pollen through water extraction and ethanol precipitation. Three homogeneous polysaccharide fractions (TPPPS1, TPPPS2, and TPPPS3) were purified from TPPPS by DEAE-cellulose column chromatography. The average molecular weights of the three polysaccharides were 56, 25, and 128 kDa, respectively. Results of high-performance liquid chromatography (HPLC) showed that TPPPS comprised mannose, ribose, xylose, glucuronic acid, galacturonic acid, glucose, galactose, and arabinose. The biological activity assays showed that TPPPS2 and TPPPS3 significantly promoted spleen lymphocyte proliferation, and that TPPPS3 showed better effect than TPPPS2. TPPPS3 enhanced the secretion of cytokine IL-2 and TNF, whereas TPPPS2 mainly elevated IL-2 secretion. By contrast, TPPPS1 exhibited other effects, and it induced the highest amount of NO production, thereby indicating that TPPPS1 had the best antioxidant activity. TPPPS3 at 50 g/mL significantly inhibited the proliferation of subgroup B Avian Leukosis virus (ALV-B) through virus adsorption interference in vitro. Results indicated that TPPPS comprised three main components, among which, TPPPS1 mainly showed antioxidant effects, whereas TPPPS2 and TPPPS3 played key roles in immunomodulation, especially TPPPS3. Further studies on the use of a reasonable proportion of TPPPS1-3 may facilitate the development of an effective immunomodulator.
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Competing Interests: The authors have declared
that no competing interests exist.
Polysaccharide, an important biomacromolecule of organisms, has a highly complex structure
and species specificity. It is a critical factor in cell surface signal identification, antigen-antibody
reaction, and intercellular signal transmission and perception [1, 2]. A growing number of
studies have corroborated that polysaccharides from natural plants could significantly improve
the function of the immune system and promote the self-protection of the body [35]. Luo
et al. reported that polysaccharides from the stems of Dendrobium officinale significantly
enhanced natural killer cell-mediated cytotoxicity and increased the phagocytosis and nitric
oxide (NO) production of macrophages [6]. Another polysaccharide from Astragalus radix
could increase the level of cytokines (TNF and GM-CSF) and the production of NO [7]. In
addition, polysaccharides from fruit calyx of Physalis alkekengi var. franchetii showed increased
scavenging effects on 1,1-dipheny-l-2-picrylhydrazyl and superoxide anion-scavenging
activities [8]. Because of their effective biological activities, plant polysaccharides have been regarded
as novel promising immunomodulatory agents, which are relatively nontoxic and with hardly
any significant side effects [911].
Pollen has been used as dietary supplement in traditional medicine for centuries, and it has
various functions, including alleviating fatigue, delaying apolexis, and treating disease. Pine
pollen, known as the King of Pollen in China, is an omnipotent nutritional pollen [12]. Oral
use of pine pollen dates back to the Tang Dynasty, which marked the beginning of the pine
pollen industry [13]. Our previous studies have demonstrated that natural nontoxic
polysaccharides derived from Taishan Pinus massoniana pollen (TPPPS) could enhance immunological
function in mice, rabbits, and chickens [1315]. Moreover, crude TPPPS, when used as an
immunoadjuvant, could significantly improve the effects of different vaccines, such as Proteus
mirabilis, the rabbit hemorrhagic disease, and the recombinant Bordetella avium ompA
subunit vaccines [13, 15, 16]. However, until now the composition of TPPPS has not been
determined, and the biological activity of each composition is also unknown.
In this study, we extracted the crude polysaccharides from Taishan P. massoniana pollen.
Subsequently, three components of TPPPS (TPPPS1, TPPPS2, and TPPPS3) with molecular
weights of 56, 25, and 128 kDa, respectively, were identified. The effects of TPPPS1-3 on spleen
lymphocyte prolifer (...truncated)