Investigation of sulfated glycosaminoglycans and their agarose gel electrophoresis patterns from plants extracts
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Published:
Jan 15, 2018
Keywords:
DMMB, uronic acids glycosaminoglycans agarose gel electrophoresis spectrophotometry
Main Article Content
Abstract
Background: Many kinds of sulfated glycosaminoglycans (GAGs) play an essential role in both physiological and pathological conditions. Most of them are obtained from animal sources, and used as nutraceuticals or therapeutic applications.
Objectives: In this study, we aimed to screen for the presence of sulfated GAGs from 8 locally available plants, including ginger (Zingiber officinale), turmeric (Curcuma longa), star fruit (Averrhoa carambola), Namwa banana (Musa ABB ‘Kluai Namwa’), bitter gourd (Momordica charantia), purple-fruited pea eggplant (Solanum trilobatum), noni (Morinda citrifolia), and finger root (Boesenbergia rotunda).
Materials and methods: All plants were extracted, and sulfated GAGs from the extracts were investigated by dimethylmethylene blue (DMMB) dye-binding assay, uronic acid assay, UV-Vis spectrophotometry, and agarose gel electrophoresis.
Results: DMMB dye-binding and uronic acids assays revealed the presence of sulfated GAGs in all extracts with various degrees of sulfated GAGs levels. These results correlated to UV-Vis spectrophotometry that showed the maximum absorbance peaks from all extracts at 190-210 nm, which was similar to sulfated GAGs standard absorption. Interestingly, agarose gel electrophoresis suggested that sulfated GAGs in all extracts exhibited diverse patterns in alcian blue, toluidine blue and safranin O staining.
Conclusion: Our results indicate that all plant extracts contain sulfated GAGs at certain levels, which could be a new approach for future study in bioprospecting.
Article Details
How to Cite
Choocheep, K., Saenkham, C., Borwornchaiyarit, S., Chaiisaraseree, K., Chimchai, J., & Moonpa, T. (2018). Investigation of sulfated glycosaminoglycans and their agarose gel electrophoresis patterns from plants extracts. Journal of Associated Medical Sciences, 51(1), 11–18. Retrieved from https://he01.tci-thaijo.org/index.php/bulletinAMS/article/view/103483
Section
Research Articles
Personal views expressed by the contributors in their articles are not necessarily those of the Journal of Associated Medical Sciences, Faculty of Associated Medical Sciences, Chiang Mai University.
References
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[26] Nathip N. Screening for glycosaminoglycans phytochemicals and antioxidant activity from local mushrooms [Term paper]. Faculty of Associated Medical Sciences: Chiang Mai University; 2014 [in Thai]
[27] Chunjarean M. Screening for phytochemicals and chondroprotective effect of garlic extracts [Term paper]. Faculty of Associated Medical Sciences: Chiang Mai University; 2015 [inThai]
[28] Nakano T, Igawa N, Ozimek L. An economical method to extract chondroitin sulphate-peptide from bovine nasal cartilage. Canadian Biosystems Engineering 2000; 42(4): 205–08.
[29] Farndale RW, Sayers CA, Barrett AJ. A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures. Connect Tissue Res 1982; 9(4): 247–8.
[30] Dietz JH, Rouse AH. A rapid method for estimating pectic substances in citrus juices. Food Res 1983; 18: 169–77.
[31] Lima MA, Rudd TR, de Farias EHC, Ebner LF, Gesteira TF, de Souza LM, et al. A new approach for heparin standardization: combination of scanning UV spectroscopy, nuclear magnetic resonance and principal component analysis. PLoS ONE 2011 Jan; 6(1): e15970. doi: 10.1371 /journal.pone.0015970.
[32] Zheng CH, Levenston ME. Fact versus artifact: avoiding erroneous estimates of sulfated glycosaminoglycan content using the dimethylmethylene blue colorimetric assay for tissue-engineered constructs. Eur Cell Mater 2015; 29: 224–36.
[33] Kubaski F, Osago H, Mason RW, Yamaguchi S, Kobayashi H, Tsuchiya M, et al. Glycosaminoglycans detection methods: Applications of mass spectrometry. Mol Genet Metab 2017 ; 120(1–2): 67–77.
[34] Stone JE. Urine analysis in the diagnosis of mucopolysaccharide disorders. Ann Clin Biochem 1998 ; 35 (Pt 2): 207–25.
[35] Stone JE, Akhtar N, Botchway S, Pennock CA. Interaction of 1,9-dimethylmethylene blue with glycosaminoglycans. Ann Clin Biochem 1994 Mar; 31 (Pt 2): 147–52.
[36] Frazier SB, Roodhouse KA, Hourcade DE, Zhang L. The Quantification of Glycosaminoglycans: A Comparison of HPLC, Carbazole, and Alcian Blue Methods. Open Glycosci 2008; 1: 31–9.
[37] Mort JS, Roughley PJ. Measurement of glycosaminoglycan release from cartilage explants. Methods Mol Med 2007; 135: 201–9.
[38] Saenkham C. Detection of phytochemicals and optimization of glycosaminoglycans by agarose gel electrophoresis in eggplant, bitter gourd, finger root and noni [Term paper]. Faculty of Associated Medical Sciences: Chiang Mai University; 2017 [in Thai]
[39] Borwornchaiyarit S. Detection of phytochemicals and optimization of glycosaminoglycans by agarose gel electrophoresis in in ginger, turmeric, star fruit and banana [Term paper]. Faculty of Associated Medical Sciences: Chiang Mai University; 2017 [in Thai]
[40] Volpi N, Maccari F. Electrophoretic approaches to the analysis of complex polysaccharides. J Chromatogr B Analyt Technol Biomed Life Sci 2006; 834(1–2): 1–13.
[41] Nader HB, Takahashi HK, Guimarães JA, Dietrich CP, Bianchini P, Ozima B. Heterogeneity of heparin: characterization of one hundred components with different anticoagulant activities by a combination of electrophoretic and affinity chromatography methods. International Journal of Biological Macromolecules 1981; 3(6): 356–60.
[42] Tomatsu S, Shimada T, Mason RW, Montaño AM, Kelly J, LaMarr WA, et al. Establishment of glycosaminoglycan assays for mucopolysaccharidoses. Metabolites 2014; 4(3): 655–79.
[43] Camplejohn KL, Allard SA. Limitations of safranin “O” staining in proteoglycan-depleted cartilage demonstrated with monoclonal antibodies. Histochemistry 1988; 89(2): 185–8.
[2] Kjellén L, Lindahl U. Proteoglycans: structures and interactions. Annu Rev Biochem 1991; 60: 443–75.
[3] Varki A, Cummings RD, Esko JD, Stanley P, Hart G, Aebi M, et al., editors. Essentials of Glycobiology [internet]. Cold Spring Harbor Laboratory Press; 2009 [cited 2017 May 30]. Available from: https://www.ncbi.nlm.nih.gov/books/NBK1900/2009.
[4] Prydz K, Dalen KT. Synthesis and sorting of proteoglycans. J Cell Sci 2000; 113 (Pt 2): 193–205.
[5] Souza-Fernandes AB, Pelosi P, Rocco PRM. Bench-to-bedside review: the role of glycosaminoglycans in respiratory disease. Crit Care 2006; 10(6): 237. doi: 10.1186/cc5069.
[6] Pomin VH. Sulfated glycans in inflammation. Eur J Med Chem 2015; 92: 353–69.
[7] Wight TN, Kang I, Merrilees MJ. Versican and the control of inflammation. Matrix Biol 2014; 35: 152–61.
[8] Kosir MA, Quinn CC, Wang W, Tromp G. Matrix glycosaminoglycans in the growth phase of fibroblasts: more of the story in wound healing. J Surg Res 2000; 92(1): 45–52.
[9] Ghatak S, Maytin EV, Mack JA, Hascall VC, Atanelishvili I, Moreno Rodriguez R, et al. Roles of Proteoglycans and Glycosaminoglycans in Wound Healing and Fibrosis. Int J Cell Biol 2015; 2015: 834893. doi: 10.1155/2015/834893
[10] Bourin MC, Lindahl U. Glycosaminoglycans and the regulation of blood coagulation. Biochem J. 1993; 289(2): 313–30.
[11] Yamada S, Sugahara K. Potential therapeutic application of chondroitin sulfate/dermatan sulfate. Curr Drug Discov Technol 2008; 5(4): 289–301.
[12] Smith RAA, Meade K, Pickford CE, Holley RJ, Merry CLR. Glycosaminoglycans as regulators of stem cell differentiation. Biochem Soc Trans 2011; 39(1): 383–7.
[13] Tanino Y, Coombe DR, Gill SE, Kett WC, Kajikawa O, Proudfoot AEI, et al. Kinetics of chemokine-glycosaminoglycan interactions control neutrophil migration into the airspaces of the lungs. J Immunol 2010; 184(5): 2677–85.
[14] Suwan K, Choocheep K, Hatano S, Kongtawelert P, Kimata K, Watanabe H. Versican/PG-M Assembles Hyaluronan into Extracellular Matrix and Inhibits CD44-mediated Signaling toward Premature Senescence in Embryonic Fibroblasts. J Biol Chem 2009; 284(13): 8596–604.
[15] Choocheep K, Hatano S, Takagi H, Watanabe H, Kimata K, Kongtawelert P, et al. Versican facilitates chondrocyte differentiation and regulates joint morphogenesis. J Biol Chem 2010; 285(27): 21114–25.
[16] Afratis N, Gialeli C, Nikitovic D, Tsegenidis T, Karousou E, Theocharis AD, et al. Glycosaminoglycans: key players in cancer cell biology and treatment. FEBS J 2012; 279(7): 1177–97.
[17] Liu D, Shriver Z, Qi Y, Venkataraman G, Sasisekharan R. Dynamic regulation of tumor growth and metastasis by heparan sulfate glycosaminoglycans. Semin Thromb Hemost 2002; 28(1): 67–78.
[18] Volpi N. Therapeutic applications of glycosaminoglycans. Curr Med Chem 2006; 13(15): 1799–810.
[19] Choi B-D, Choi YJ. Nutraceutical functionalities of polysaccharides from marine invertebrates. Adv Food Nutr Res 2012; 65: 11–30.
[20] Ronca G, Conte A. Metabolic fate of partially depolymerized shark chondroitin sulfate in man. Int J Clin Pharmacol Res 1993; 13 (Suppl): S27–34.
[21] Uebelhart D. Clinical review of chondroitin sulfate in osteoarthritis. Osteoarthr Cartil 2008; 16 (Suppl 3): S19-21.
[22] Volpi N. Oral bioavailability of chondroitin sulfate (Condrosulf) and its constituents in healthy male volunteers. Osteoarthr Cartil 2002; 10(10): 768–77.
[23] Kubo M, Ando K, Mimura T, Matsusue Y, Mori K. Chondroitin sulfate for the treatment of hip and knee osteoarthritis: current status and future trends. Life Sci 2009; 85(13–14): 477–83.
[24] Linhardt RJ, Gunay NS. Production and chemical processing of low molecular weight heparins. Semin Thromb Hemost 1999; 25 (Suppl 3): S5–16.
[25] Gandhi NS, Mancera RL. The structure of glycosaminoglycans and their interactions with proteins. Chem Biol Drug Des 2008; 72(6): 455–82.
[26] Nathip N. Screening for glycosaminoglycans phytochemicals and antioxidant activity from local mushrooms [Term paper]. Faculty of Associated Medical Sciences: Chiang Mai University; 2014 [in Thai]
[27] Chunjarean M. Screening for phytochemicals and chondroprotective effect of garlic extracts [Term paper]. Faculty of Associated Medical Sciences: Chiang Mai University; 2015 [inThai]
[28] Nakano T, Igawa N, Ozimek L. An economical method to extract chondroitin sulphate-peptide from bovine nasal cartilage. Canadian Biosystems Engineering 2000; 42(4): 205–08.
[29] Farndale RW, Sayers CA, Barrett AJ. A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures. Connect Tissue Res 1982; 9(4): 247–8.
[30] Dietz JH, Rouse AH. A rapid method for estimating pectic substances in citrus juices. Food Res 1983; 18: 169–77.
[31] Lima MA, Rudd TR, de Farias EHC, Ebner LF, Gesteira TF, de Souza LM, et al. A new approach for heparin standardization: combination of scanning UV spectroscopy, nuclear magnetic resonance and principal component analysis. PLoS ONE 2011 Jan; 6(1): e15970. doi: 10.1371 /journal.pone.0015970.
[32] Zheng CH, Levenston ME. Fact versus artifact: avoiding erroneous estimates of sulfated glycosaminoglycan content using the dimethylmethylene blue colorimetric assay for tissue-engineered constructs. Eur Cell Mater 2015; 29: 224–36.
[33] Kubaski F, Osago H, Mason RW, Yamaguchi S, Kobayashi H, Tsuchiya M, et al. Glycosaminoglycans detection methods: Applications of mass spectrometry. Mol Genet Metab 2017 ; 120(1–2): 67–77.
[34] Stone JE. Urine analysis in the diagnosis of mucopolysaccharide disorders. Ann Clin Biochem 1998 ; 35 (Pt 2): 207–25.
[35] Stone JE, Akhtar N, Botchway S, Pennock CA. Interaction of 1,9-dimethylmethylene blue with glycosaminoglycans. Ann Clin Biochem 1994 Mar; 31 (Pt 2): 147–52.
[36] Frazier SB, Roodhouse KA, Hourcade DE, Zhang L. The Quantification of Glycosaminoglycans: A Comparison of HPLC, Carbazole, and Alcian Blue Methods. Open Glycosci 2008; 1: 31–9.
[37] Mort JS, Roughley PJ. Measurement of glycosaminoglycan release from cartilage explants. Methods Mol Med 2007; 135: 201–9.
[38] Saenkham C. Detection of phytochemicals and optimization of glycosaminoglycans by agarose gel electrophoresis in eggplant, bitter gourd, finger root and noni [Term paper]. Faculty of Associated Medical Sciences: Chiang Mai University; 2017 [in Thai]
[39] Borwornchaiyarit S. Detection of phytochemicals and optimization of glycosaminoglycans by agarose gel electrophoresis in in ginger, turmeric, star fruit and banana [Term paper]. Faculty of Associated Medical Sciences: Chiang Mai University; 2017 [in Thai]
[40] Volpi N, Maccari F. Electrophoretic approaches to the analysis of complex polysaccharides. J Chromatogr B Analyt Technol Biomed Life Sci 2006; 834(1–2): 1–13.
[41] Nader HB, Takahashi HK, Guimarães JA, Dietrich CP, Bianchini P, Ozima B. Heterogeneity of heparin: characterization of one hundred components with different anticoagulant activities by a combination of electrophoretic and affinity chromatography methods. International Journal of Biological Macromolecules 1981; 3(6): 356–60.
[42] Tomatsu S, Shimada T, Mason RW, Montaño AM, Kelly J, LaMarr WA, et al. Establishment of glycosaminoglycan assays for mucopolysaccharidoses. Metabolites 2014; 4(3): 655–79.
[43] Camplejohn KL, Allard SA. Limitations of safranin “O” staining in proteoglycan-depleted cartilage demonstrated with monoclonal antibodies. Histochemistry 1988; 89(2): 185–8.