Characterization and Biological Activity of Taishan Pinus massoniana Pollen Polysaccharide In Vitro

PLOS ONE, Dec 2019

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 μg/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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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. - 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)


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Shifa Yang, Kai Wei, Fengjuan Jia, Xue Zhao, Guolin Cui, Fanxia Guo, Ruiliang Zhu. Characterization and Biological Activity of Taishan Pinus massoniana Pollen Polysaccharide In Vitro, PLOS ONE, 2015, 3, DOI: 10.1371/journal.pone.0115638