Characterization and antioxidant activities of the polysaccharides from Radix Cyathulae officinalis Kuan

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Abstract

A water-soluble polysaccharide (PSRC) was extracted and purified from the roots of Radix Cyathulae officinalis Kuan, and its chemical characteristics, monosaccharide composition and antioxidant activities were characterized. The average of molecular weight (Mw) of PSRC was 182 kDa. The majority of monosaccharide components of PSRC was glucose (relative mass 48.8%) with lower levels of mannose, rhamnose, galactose, fructose and arabinose (relative mass of 15.7, 14.3, 10.6, 6.1 and 4.5%, respectively). In vitro assays revealed that RSRC possessed potent scavenging activities against DPPH, hydroxyl and superoxide anion radicals. Oral administration of PSRC significantly enhanced antioxidant enzyme activities (including total superoxide dismutase, copper-zinc superoxide dismutase (Cu,Zn-SOD), manganese superoxide dismutase (Mn-SOD), glutathione peroxidase and catalase (CAT)) and capacities of scavenging superoxide anion and hydroxyl radicals, markedly lowered lipid peroxidation formation of malondialdehyde and significantly up-regulated mRNA expressions of Cu,Zn-SOD, Mn-SOD, CAT, glutathione peroxidase 1, thioredoxin 1 and thioredoxin 2, in a d-galactose-induced aging mouse model. Taken together, these findings demonstrate that PSRC could be used as a novel promising source of natural antioxidants and antiaging drugs.

Introduction

Oxidizing reactions occur constantly in the cells as part of aerobic life, resulting in the production of oxygen-derived radicals (reactive oxygen species (ROS)). ROS, if produced in excess, may directly damage various organic substrates, including DNA, proteins, lipids and cell membranes in living cells, and/or indirectly act as primary or secondary messengers to activate signaling pathways that inflict damage on living cells [1], thus resulting in various diseases and disorders, such as cardiovascular diseases, autoimmune disorders, cancer, rheumatoid arthritis and aging [2]. Therefore, it is essential to develop and utilize effective antioxidants to protect living organisms against oxidative damage caused by ROS. Several synthesized commercial antioxidants have been widely used in resisting oxidative damages [3]. However, the presence of undesired side effects from these synthesized antioxidants is almost unavoidable, and those most commonly observed have been suspected of being responsible for liver damage and carcinogenicity [3]. Therefore, it is essential to develop natural antioxidants with low or even no cytotoxicity.

In recent years, amounting studies have documented that polysaccharides from plants, especially from the herbal plants, possess potent antioxidant but low cytotoxic activities, and thus could be explored as sources of novel potential antioxidants [4], [5]. However, most of the previous studies evaluating the antioxidant potential of polysaccharides from plants were just carried out in various in vitro systems, while further in vivo evaluations were rarely performed.

Radix Cyathulae officinalis Kuan, belonging to Amaranthaceae family, is mainly distributed in Sichuan province, China. The roots of Cyathula officinalis Kuan (“Chuan Niu Xi” in Chinese, C. officinalis Kuan) have been included in the Pharmocopeia of the People's Republic of China due to its various pharmacological activities, including analgesic, immunostimulant, antitumor, anti-inflammatory, antiaging, removing blood stasis, restoring menstrual flow, inducing diuresis for treating stranguria [6]. Various biologically active compounds, such as phytoecdysteroids, palmitic acids and hyterocyclic compounds [7], [8], have been isolated from C. officinalis Kuan, and their biological properties have been characterized. However, a little attention was devoted to the isolation and identification of polysaccharides from C. officinalis Kuan. Moreover, the antioxidant properties of polysaccharides from C. officinalis Kuan have not been characterized yet.

In this work, a novel polysaccharide was extracted and purified from C. officinalis Kuan. Its chemical characteristics, monosaccharide composition and structural property were identified, and its antioxidant activities both in vitro and in vivo were evaluated for the first time.

Section snippets

Chemicals and reagents

2,2-Diphenyl-1-picryl-hydrazyl (DPPH), vitamin C (Vc), glucuronic acid and serial standard monosaccharides (d-glucose, d-xylose, d-fructose, d-galactose, l-rhamnose, d-mannose and d-arabinose) were purchased Sigma Chemical Co. (St. Louis, Mo, USA). Dextrans of different molecular weights were from Pharmacia Co. (Uppsala, Sweden). The commercial assay kits for hydroxyl and superoxide radicals and antioxidant enzymes (including total superoxide dismutase, copper-zinc superoxide dismutase,

Isolation, purification, and chemical properties of polysaccharide

The crude polysaccharide (APPC) extracted from C. officinalis Kuan was obtained and presented negative iodine-potassium iodide reaction, indicating the absence of starch-type polysaccharide. APPC was purified by DEAE-Sepharose Fast-flow column chromatography. Three fractions were isolated (data were not shown), and the main fraction, which eluted with 0.4 M NaCl solution, was collected and designed as PSRC. PSRC appeared as white-colored power. The chemical characteristics and monosaccharide

Discussion

In present study, a soluble polysaccharide (PSRC) from Radix Cyathulae officinalis Kuan was extracted and purified, and its chemical characteristics and monosaccharide composition, as well as in vitro and in vivo antioxidant activities were investigated. The FT-IR spectrum revealed obvious characteristic absorption peaks of polysaccharide, and the presence of both α- and β-type glycosidic bond in their structure. GC-MS analysis shown that PSRC was consisted of glucose, rhamnose, mannose,

Acknowledgements

This work was supported in part by Sichuan Agricultural University Excellent Doctoral Dissertation program (YB2014004), Two Sides Supporting plan in Sichuan Agricultural University (00770107), and Sichuan Science and Technology Commission (Project No. 2012HH0013).

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