Antioxidant and immonomodulatory activities of polysaccharides isolated from Taxillus Chinensis and Uncaria rhyncophylla Lin Zhang, Narsimha Reddy, Sundar Rao Koyyalamudi and Sang Chul Jeong 1

Background Cancer Treatment According to the World Health Organisation (WHO), cancer killed more than 11.5 millions people around the world in 2030 Treatment Western Medicine is Expensive - Not accessible for majority Traditional Medicine is cost effective - More research needed 2

Background Botanical Polysaccharides Non-toxicity Minimal Side- effects Immunomodulatory properties Anti-tumour actitive Joeng et al., 2012 J. Med. Food, 15, 58-65. Zhang et al., 2012, Int. J. Biol. Maromol (In press) 3

Aim and objective Aim : Objectives : Isolation and measurement of immunomodulatory activities of polysaccharides from Uncaria rhyncophylla and Taxillus chinensis Objectives : Factionation of bioactive polysaccharides from U. rhyncophylla and T. chinensis using gel filtration column Screening these fractions for their immunomodulatory effects 4

Crude polysaccharides Methodology Hot water extraction (autoclaving) (121oC, 2h) EtOH precipitation (95%) (Supernatant:EtOH =1:4 /24h) Centrifugation (10,447xg / 20min) Centrifugation (10,447xg / 20min) Gel chromatography Sepharose CL-6B Phenol-sulfuric method Medicinal plant Supernatant Precipitate Crude polysaccharides Different fractions Joeng et al., 2012 J. Med. Food, 15, 58-65. 5

Results of fractionation of polysaccharide Uncaria Taxillus 6

The polysaccharide fractions of Taxillus chinensis: TCP-1 TCP-2 TCP-3 The polysaccharide fractions of Uncaria rhyncophylla: UC-1 UC-2 7

Preliminary characterisation Chemical composition analysis - Analysis of mono-saccharides - Mainly contains Glucose, Fructose, Xylose, Arabinose and Rhamnose 8

Immunomodulatory and Antioxidant activities - Antioxidant assays -DPPH- free radical test -Yeast oxidative stress assay - Immunomodulatory properties - NO production - TNF-α production - Cell viability 9

Results: Antioxidant activity   % Yeast cell growth (Water extracts) * S.NO (1mg/ml) Molecular Mass (kDa) DPPH scavenging activity of water extracts (Ascorbate equivalent µM) # CHP (150 µM) H2O2 (2mM) LAH (75 µM) TCP-1 541 207 ± 4 27.79 ± 2.11 23.02 ± 1.94 TCP-2 18 447 ± 0 30 ± 3.54 39.7 ± 2.11 25.09 ± 1.46 TCP-3 2 662 ± 0 37.5 ± 0 48.21 ± 2.11 32.65 ± 3.89 UC-1 524 245 ± 0 45 ± 3.54 41.17 ± 4.21 22.68 ± 0.49 UC-2  3 305 ± 0 77.5 ± 7.07 70.52 ± 2.11 25.09 ± 0.49 10

Results: Immunomodulatory properties (T. chinensis) 11

Results: Immunomodulatory properties (U. rhynchophylla) 4/25/2017 12

Results of cell viabilities 13

Conclusion Polysaccharides of T. chinensis and U. rhyncophylla displayed high immunomodulatory activities Huge potential of these polysaccharides as natural anti-tumor agents Further studies 14

References Schepetkin, I.A. & Quinn, M.T. Botanical polysaccharides: macrophage immunomodulation and therapeutic potential, Immunopharmacology 6 (2006) 317-333. Jeong, S.C., Koyyalamudi, S.R., Jeong, Y.T., Song, C.H., & Pang, G. (2012). Macrophage Immunomodulating and Antitumor Activities of Polysaccharides Isolated from Agaricus bisporus White Button Mushrooms, Journal of Medicinal Food, 15, 58-65. Sun, L., Peng, X., Sun, P., Shi, J., Yuan, X., Zhu, J., Tai, G., & Zhou, Y. (2012). Structural characterization and immunostimulatory activity of a novel linear α- (1→6)-D-glucan isolated from Panax ginseng CA Meyer. Glycoconjugate Journal, 1-8. Jeong, S.C., Jeong, Y.T., Yang, B.K., Islam, R., Koyyalamudi, S.R., Pang, G., Cho, K.Y., & Song, C.H. (2010). White button mushroom (Agaricus bisporus) lowers blood glucose and cholesterol levels in diabetic and hypercholesterolemic rats, Nutrition Research, 30, 49-56 15

Acknowledgements First and foremost, I would like to thank you my supervisor, Dr. Smith, Dr.Reddy and Dr.Rao for their valuable guidance and advice. I also thank Dr. Jeong for helping me in biological experiments. The generous support of technical staff is also acknowledged. 16

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