Comparative analysis of structural characteristics and cardiomyocyte protective effects of glucomannan extracted from Bletilla striata using varied methods

doi:10.5246/jcps.2024.08.051

Abstract

Abstract:

Doxorubicin (DOX) is a widely employed tumor therapy, yet its substantial toxic side effects pose a considerable challenge. Bletilla striata has demonstrated efficacy in preventing and treating these toxic side effects in clinical practice, with polysaccharides identified as the principal active component. In the present study, 16 fractions of B. striata polysaccharides (BsPs) were extracted using diverse methods, including hot-water extraction (HWE), ultrasonic-assisted extraction (UAE), enzyme-assisted extraction (EAE), dilute acid-water extraction (ACWE), and dilute alkali-water extraction (ALWE). These extractions were subsequently precipitated at final ethanol concentrations of 80% and 95%, respectively. The investigation encompassed yields, total carbohydrate content (TCC), total protein content (TPC), preliminary structural properties, and anti-DOX myocardial cytotoxic activity. Results indicated that the extraction method significantly influenced the physicochemical properties, associated functional properties, and anti-DOX myocardial cytotoxic activity of BsPs. HWE and UAE yielded higher BsPs quantities. The relative molecular weight (RMW) distribution of BsPs differed notably between HWE or UAE, EAE, ACWE, and ALWE. The RMW of primary BsPs obtained from HWE and UAE (1.9 × 10⁷–1.7 × 10⁷ Da) exceeded that from EAE (7.5 × 10³–2.8 × 10⁴ Da) and ALWE (5.1 × 10⁴–1.7 × 10⁴ Da), with smaller molecular weights primarily precipitated by higher ethanol concentrations. BsPs were composed of Man and Glu, with partial fractions containing small amounts of Gal or Ara, displaying varying molar ratios. Notably, BsPs from ACWE exhibited the most significant structural differences, lacking 1,4-α-D-Glcp and a triple-helical structure. Furthermore, BsPs obtained from HWE, UAE, and EAE demonstrated heightened anti-DOX myocardial cytotoxic activity compared to other methods. This study underscored the influence of extraction methods on BsPs’ structure and myocardial protective activity, offering a foundation for exploring structural diversity and employing specific extraction methods to extract polysaccharides with robust myocardial protective properties efficiently.

Key words: Bletilla striata (Thunb.) Rchb. f., Polysaccharide, Extraction methods, Structural characteristics, Myocardial protection

Supporting:

Feng Pan, Nan Yang, Guoyong Ruan, Changyan Yu, Jianbo Yang, Dongxue Suo, Ying Qin, Yun Liu. Comparative analysis of structural characteristics and cardiomyocyte protective effects of glucomannan extracted from Bletilla striata using varied methods[J]. Journal of Chinese Pharmaceutical Sciences, 2024, 33(8): 686-704.

Figures/Tables 11

Table 1. The RMW of BsPs calculated based on their RT values obtained from high-performance gel permeation chromatograms.

Table 2. The molar ratio of monosaccharide composition for BsPs extracted by different extraction methods.

References 45

[1]	China P.C. Pharmacopoeia of the People’s Republic of China: Chemical Industry Press. 2020.
[2]	Gou, K.J.; Li, Y.; Qu, Y.; Li, H.R.; Zeng, R. Advances and prospects of Bletilla striata polysaccharide as promising multifunctional biomedical materials. Mater. Des. 2022, 223, 111198.
[3]	Yu, Y.; Shen, M.Y.; Song, Q.Q.; Xie, J.H. Biological activities and pharmaceutical applications of polysaccharide from natural resources: a review. Carbohydr. Polym. 2018, 183, 91–101.
[4]	Chen, Z.Y.; Cheng, L.Z.; He, Y.C.; Wei, X.L. Extraction, characterization, utilization as wound dressing and drug delivery of Bletilla striata polysaccharide: a review. Int. J. Biol. Macromol. 2018, 120, 2076–2085.
[5]	Zhang, W.; Ma, S.; Gu, G.; Shi, J.; Zhao, B. Studies on the toxicological assessment on skin safety of polysaccharide gum of Bletilla striata (Thunb.) Reichb. f. Chin. Wild Plant Res. 2003, 22, 59–61.
[6]	Zhou, P.; Zhao, S.Y.; Huang, C.; Qu, Y.; Zhang, C. Bletilla striata polysaccharide microneedle for effective transdermal administration of model protein antigen. Int. J. Biol. Macromol. 2022, 205, 511–519.
[7]	Guan, H.Y.; Yan, Y.; Wang, Y.L.; Wang, A.M.; Liu, J.H.; He, X.; Li, Y.J.; Huang, Y.; Liao, S.G. Isolation and characterization of two new 2-isobutylmalates from Bletilla striata. Chin. J. Nat. Med. 2016, 14, 871–875.
[8]	Xu, D.L.; Pan, Y.C.; Chen, J.S. Chemical constituents, pharmacologic properties, and clinical applications of Bletilla striata. Front. Pharmacol. 2019, 10, 1168.
[9]	Ma, Y.N.; He, S.L.; Ma, X.Q.; Hong, T.T.; Li, Z.F.; Park, K.; Wang, W.P. Silymarin-loaded nanoparticles based on stearic acid-modified Bletilla striata polysaccharide for hepatic targeting. Molecules. 2016, 21, 265.
[10]	Mi, Z.Y.; Lv, D.H.; Jiang, G.H.; Niu, J.F.; Wang, S.Q.; Wang, Z.Z. Establishment of a rapid breeding system for Bletilla striata. HortScience. 2021, 56, 454–459.
[11]	Xu, Z.; Feng, S.L.; Qu, J.P.; Yuan, M.; Yang, R.W.; Zhou, L.J.; Chen, T.; Ding, C.B. The effect of extraction methods on preliminary structural properties and antioxidant activities of polysaccharides from lactarius vividus. Processes. 2019, 7, 482.
[12]	Wang, C.M.; Sun, J.T.; Luo, Y.; Xue, W.H.; Diao, H.J.; Dong, L.; Chen, J.N.; Zhang, J.F. A polysaccharide isolated from the medicinal herb Bletilla striata induces endothelial cells proliferation and vascular endothelial growth factor expression in vitro. Biotechnol. Lett. 2006, 28, 539–543.
[13]	Wang, Y.; Liu, D.; Chen, S.J.; Wang, Y.; Jiang, H.X.; Yin, H.P. A new glucomannan from Bletilla striata: structural and anti-fibrosis effects. Fitoterapia. 2014, 92, 72–78.
[14]	Peng, Q.; Li, M.; Xue, F.; Liu, H.J. Structure and immunobiological activity of a new polysaccharide from Bletilla striata. Carbohydr. Polym. 2014, 107, 119–123.
[15]	Wang, B.; Sha, X.U.; Huang, L.J.; Wang, Z.F. Isolation, purification and structural characterization of a polysaccharide fraction from stem tuber of Bletilla striata, named BSPI-A. Food Sci. 2010, 31, 120–123.
[16]	Wang, Y.R.; Han, S.W.; Li, R.F.; Cui, B.S.; Ma, X.J.; Qi, X.Z.; Hou, Q.; Lin, M.B.; Bai, J.Y.; Li, S.A. Structural characterization and immunological activity of polysaccharides from the tuber of Bletilla striata. Int. J. Biol. Macromol. 2019, 122, 628–635.
[17]	Chen, Z.Y.; Zhao, Y.; Zhang, M.K.; Yang, X.F.; Yue, P.X.; Tang, D.K.; Wei, X.L. Structural characterization and antioxidant activity of a new polysaccharide from Bletilla striata fibrous roots. Carbohydr. Polym. 2020, 227, 115362.
[18]	Liu, C.; Dai, K.Y.; Ji, H.Y.; Jia, X.Y.; Liu, A.J. Structural characterization of a low molecular weight Bletilla striata polysaccharide and antitumor activity on H22 tumor-bearing mice. Int. J. Biol. Macromol. 2022, 205, 553–562.
[19]	Jiang, G.H.; Wang, B.L.; Wang, Y.W.; Kong, H.Y.; Wang, Y.F.; Gao, P.; Guo, M.H.; Li, W.N.; Zhang, J.; Wang, Z.Z.; Niu, J.F. Structural characteristics of a novel Bletilla striata polysaccharide and its activities for the alleviation of liver fibrosis. Carbohydr. Polym. 2023, 313, 120781.
[20]	Cagel, M.; Grotz, E.; Bernabeu, E.; Moretton, M.A.; Chiappetta, D.A. Doxorubicin: nanotechnological overviews from bench to bedside. Drug Discov. Today. 2017, 22, 270–281.
[21]	Chen, S.; Sutiman, N.; Zhang, C.Z.; Yu, Y.N.; Lam, S.; Khor, C.C.; Chowbay, B. Pharmacogenetics of irinotecan, doxorubicin and docetaxel transporters in Asian and Caucasian cancer patients: a comparative review. Drug Metab. Rev. 2016, 48, 502–540.
[22]	Page, R.L. II, O’Bryant, C.L.; Cheng, D.; Dow, T.J.; Ky, B.; Stein, C.M.; Spencer, A.P.; Trupp, R.J.; Lindenfeld, J. Drugs that may cause or exacerbate heart failure. Circulation. 2016, 134, e32–e69.
[23]	Yang, J. Prevention and treatment of postoperative vomiting in 30 cases of liver interventional surgery with Bletilla striata. J. Tradit. Chin. Med. 2002, 43, 611–612.
[24]	He, F.L. BCA (bicinchoninic acid) protein assay. Bio-protocol. 2011, 1, e44.
[25]	Liu, J.; Luo, J.G.; Sun, Y.; Ye, H.; Lu, Z.X.; Zeng, X.X. A simple method for the simultaneous decoloration and deproteinization of crude levan extract from Paenibacillus polymyxa EJS-3 by macroporous resin. Bioresour. Technol. 2010, 101, 6077–6083.
[26]	Li, N.S.; Yan, C.Y.; Hua, D.H.; Zhang, D.Z. Isolation, purification, and structural characterization of a novel polysaccharide from Ganoderma capense. Int. J. Biol. Macromol. 2013, 57, 285–290.
[27]	Pan, F.; Hou, K.; Li, D.D.; Su, T.J.; Wu, W. Exopolysaccharides from the fungal endophytic Fusarium sp. A14 isolated from Fritillaria unibracteata Hsiao et KC Hsia and their antioxidant and antiproliferation effects. J. Biosci. Bioeng. 2019, 127, 231–240.
[28]	Yang, N.; Ruan, G.Y.; Wang, C.Y.; Yang, J.; Liu, Y.; Xie, F.Y.; Li, C.J.; Pan, F. Study on simultaneous decoloration and deproteinization of crudepolysaccharide from the root of Caesalpinia decapetala using macroporous adsorption resin. J. Guizhou Normal. Univ. 2023, 41, 107–113.
[29]	Papazisis, K.T.; Geromichalos, G.D.; Dimitriadis, K.A.; Kortsaris, A.H. Optimization of the sulforhodamine B colorimetric assay. J. Immunol. Methods. 1997, 208, 151–158.
[30]	Chen, X.H.; Chen, G.J.; Wang, Z.R.; Kan, J.Q. A comparison of a polysaccharide extracted from ginger (Zingiber officinale) stems and leaves using different methods: preparation, structure characteristics, and biological activities. Int. J. Biol. Macromol. 2020, 151, 635–649.
[31]	Wang, S.B.; Qin, X.M.; Guo, X.Q.; Wang, G.L.; Zhang, L.Z. Decolorization of crude saposhnikovia divaricata polysaccharide by resins. Chin. J. Applied Chem. 2005, 22, 1308–1311.
[32]	Chen, G.J.; Fang, C.C.; Ran, C.X.; Tan, Y.; Yu, Q.Q.; Kan, J.Q. Comparison of different extraction methods for polysaccharides from bamboo shoots (Chimonobambusa quadrangularis) processing by-products. Int. J. Biol. Macromol. 2019, 130, 903–914.
[33]	Chen, X.; Zhang, H.B.; Du, W.Q.; Qian, L.Y.; Xu, Y.; Huang, Y.G.; Xiong, Q.P.; Li, H.L.; Yuan, J. Comparison of different extraction methods for polysaccharides from Crataegus pinnatifida Bunge. Int. J. Biol. Macromol. 2020, 150, 1011–1019.
[34]	Jiang, Y.P.; Qi, X.H.; Gao, K.; Liu, W.J.; Li, N.; Cheng, N.B.; Ding, G.; Huang, W.Z.; Wang, Z.Z.; Xiao, W. Relationship between molecular weight, monosaccharide composition and immunobiologic activity of Astragalus polysaccharides. Glycoconj. J. 2016, 33, 755–761.
[35]	Pan, F.; Su, T.J.; Liu, Y.; Hou, K.; Chen, C.; Wu, W. Extraction, purification and antioxidation of a polysaccharide from Fritillaria unibracteata var. wabuensis. Int. J. Biol. Macromol. 2018, 112, 1073–1083.
[36]	Zhang, C.; Gao, F.; Gan, S.; He, Y.N.; Chen, Z.J.; Liu, X.W.; Fu, C.M.; Qu, Y.; Zhang, J.M. Chemical characterization and gastroprotective effect of an isolated polysaccharide fraction from Bletilla striata against ethanol-induced acute gastric ulcer. Food Chem. Toxicol. 2019, 131, 110539.
[37]	Ayimbila, F.; Keawsompong, S. Functional composition and antioxidant property of crude polysaccharides from the fruiting bodies of Lentinus squarrosulus. Biotech. 2021, 11, 7.
[38]	Chen, H.Y.; Zeng, J.S.; Wang, B.; Cheng, Z.; Xu, J.; Gao, W.H.; Chen, K.F. Structural characterization and antioxidant activities of Bletilla striata polysaccharide extracted by different methods. Carbohydr. Polym. 2021, 266, 118149.
[39]	Hu, B.F.; Yang, H.B.; Chen, G.M.; Sun, X.J.; Zou, X.J.; Ma, J.; Yao, X.W.; Liang, Q.; Liu, H.T. Structural characterization and preventive effect on non-alcoholic fatty liver disease of oligosaccharides from Bletilla striata. Food Funct. 2022, 13, 4757–4769.
[40]	Jia, X.J.; Hu, J.; He, M.X.; Zhang, Q.W.; Li, P.; Wan, J.B.; He, C.W. α-Glucosidase inhibitory activity and structural characterization of polysaccharide fraction from Rhynchosia minima root. J. Funct. Foods. 2017, 28, 76–82.
[41]	Niu, J.F.; Wang, S.P.; Wang, B.L.; Chen, L.J.; Zhao, G.M.; Liu, S.; Wang, S.Q.; Wang, Z.Z. Structure and anti-tumor activity of a polysaccharide from Bletilla ochracea schltr. Int. J. Biol. Macromol. 2020, 154, 1548–1555.
[42]	Mondal, S.; Chakraborty, I.; Pramanik, M.; Rout, D.; Islam, S.S. Structural studies of an immunoenhancing polysaccharide isolated from mature pods (fruits) of moringa oleifera (sajina). Med. Chem. Res. 2004, 13, 390–400.
[43]	Jones, C.; Lemercinier, X. Full NMR assignment and revised structure for the capsular polysaccharide from Streptococcus pneumoniae type 15B. Carbohydr. Res. 2005, 340, 403–409.
[44]	Shang, H.M.; Zhou, H.Z.; Li, R.; Duan, M.Y.; Wu, H.X.; Lou, Y.J. Extraction optimization and influences of drying methods on antioxidant activities of polysaccharide from cup plant (Silphium perfoliatum L.). PLoS One. 2017, 12, e0183001.
[45]	Qu, Y.; Li, C.X.; Zhang, C.; Zeng, R.; Fu, C.M. Optimization of infrared-assisted extraction of Bletilla striata polysaccharides based on response surface methodology and their antioxidant activities. Carbohydr. Polym. 2016, 148, 345–353.