BIOLOGICAL ACTIVITY OF BULGARIAN FOLIA BETULAE DRY EXTRACT
Keywords:
Folia Betulaeextract, HPLC identification, Biological activityAbstract
Objective: The aim of this study was to investigate the biological activity of dry Folia Betulae (FB) extract.
Methods: Extracts from birch leaves were obtained by different technological methods–maceration and percolation, extraction with different concentrations of ethanol, changes in temperature regimen. The influence of the technological factors on the content of the biologically active substances (BAS) was examined. A phytochemical characterization of the extracts and their standardization were made, according to important groups of BAS–flavonoids (rutin, quercetin) and terpenes (betulin and betulinic acid), by means of HPLC methods for detection and quantitative determination. A model extract, with optimal content of BAS was chosen for subsequent in vitro investigation of its biological activity. Antimicrobial activity was studied via in vitro tests using bacterial isolates–Staphylococcus aureus, Escherichia coli and Candida albicans. The physiological activity was investigated by using in vitro test with smooth muscle strips. The antiproliferative activity of FB extract on eukaryotic cells was examined on cell cultures in vitro. Two cell cultures were used: the mouse lymphoma cell line L5178Y and the serum-free McCoy-Plovdiv cells.
Results: The dry extract from FB has a dose–dependent antibacterial effect. The bactericidal effect on Staphylococcus aureus is stronger than the one on Escherichia coli. Results prove that adding the extract leads to stimulating effect on muscle contractility. It demonstrates biological activity, expressed as changes in cell morphology, proliferation and vitality as well as initiation of apoptosis.
Conclusion: The results obtained largely overlap with literature data and explain some of the applications of this plant in traditional medicine.
Â
Downloads
References
Klinger W, Hirschelmann R, Süss J. Birch sap and birch leaves extract: screening for antimicrobial, phagocytosis-influencing, antiphlogistic and antipyretic activity. Pharmazie 1989; 44(8):558-60.
Kandaswami C, Lee LT, Lee PP, Hwang JJ, Ke FC, Huang YT, et al. The antitumor activities of flavonoids. In Vivo 2005;19:895-909.
Drag M, Surowiak P, Drag-Zalesinska M, Dietel M, Lage H, Oleksyszyn J. Comparision of the cytotoxic effects of birch bark extract, Betulin and Betulinic acid towards human gastric carcinoma and pancreatic carcinoma drug-sensitive and drug-resistant cell lines. Molecules 2009;14(4):1639-51.
Ju EM, Lee SE, Hwang HJ, Kim JH. Antioxidant and anticancer activity of extract from Betula platyphylla var. japonica. Life Sci 2004;74:1013-26.
Smullen J, Koutsou G, Foster H, Zumbe A, Storey D. The antibacterial activity of plant extracts containing polyphenols against Streptococcus mutans. Caries Res 2007;41(5):342-9.
Cushnie ТP, Lamb ÐJ. Antimicrobial activity of flavonoids. Int J Antimicrob Agents 2005;26(5):343-56.
Pepeljnjak S, Kalodera Z, Zovko M. Antimicrobial activity of flavonoids from Pelargonium radula (Cav.) L’Herit. Acta Pharm 2005;55(4):431-5.
Akiyama H, Fujii K, Yamasaki O, Oono T, Iwatsuki K. Antibacterial action of several tannins against Staphylococcus aureus. J Antimicrob Chemother 2001;48(4):487-91.
Webster D, Taschereau P, Belland RJ, SC Rennie RP. Antifungal activity of medicinal plant extracts; preliminary screening studies. J Ethnopharmacol 2008;115:140-6.
Qi-he C, Jing L, Hai-feng Z, Guo-qing H, Ming-liang F. The betulinic acid production from betulin through biotransformation by fungi. Enzyme Microb Technol 2009;45(3):175–80.
Grassman J. Terpenoids as plant antioxidants. Vitam Horm 2005;72:505-35
Prokof'eva NG, Anisimov MM, Kiseleva MI, Rebachuk NM, Pokhilo ND. Cytotoxic activity of dammarane triterpenoids from birch leaves. Izv Akad Nauk Ser Biol 2002;6:645-9.
Setzer WN, Setzer MC. Plant-derived triterpenoids as potential antineoplastic agents. Mini Rev Med Chem 2003;3(6):540-56.
Rzeski W, Stepulak A, Szymański M, Sifringer M, Kaczor J, Wejksza K, et al. Betulinic acid decreases expression of bcl-2 and cyclin D1, inhibits proliferation, migration and induces apoptosis in cancer cells. Naunyn Schmiedebergs Arch Pharmacol 2006;374(1):11-20.
Bishayee A, Ahmed S, Brankov N, Perloff M. Triterpenoids as potential agents for the chemoprevention and therapy of breast cancer. Front Biosci 2011;16:980–96.
Zhao G, Yan W, Cao D. Simultaneous determination of betulin and betulinic acid in white birch bark using RP-HPLC. J Pharm Biomed Anal 2007;43(3):959–62.
Abreu I, Porto A, Marsaioli A, Mazzafera P. Distribution of bioactive substances from Hypericum brasiliense during plant growth. Plant Sci 2004;167(4):949–54.
Zu Y, Li C, Fu Y, Zhao C. Simultaneous determination of catechin, rutin, quercetin kaempferol and isorhamnetin in the extract of sea buckthorn (Hippophae rhamnoides L.) leaves by RP-HPLC with DAD. J Pharm Biomed Anal 2006;41(3):714-9.
Demirci F, Demirci B, Baser KHC, Güven K. The composition and antifungal bioassay of the essential oils of different Betula species growing in Turkey. Chem Nat Compd 2000;36(2):159-65.
Shimizu I, Isshiki Y, Nomura H, Sakuda K, Sakuma K, Kondo S. The antibacterial activity of fragrance ingredients against Legionella pneumophila. Biol Pharm Bull 2009;32(6):1114-7.
Demikhova OV, Balakshin VV, Presnova GA, Bocharova IV, Lepekha LN, Chernousova LN, et al. Antimycobacterial activity of a dry birch bark extract on a model of experimental pulmonary tuberculosis. Probl Tuberk Bolezn Legk 2006;(1):55-7.
Berridge MJ. Cell Signalling Pathways. In: Cell Signalling Biology, Module 2; 2012.
http://www.auburn.edu/academic/classes/biol/6190/CellSignalingBiology/csb002. pdf
Ratz PH, Berg KM, Urban NH, Miner AS. Regulation of smooth muscle calcium sensitivity: KCl as a calcium-sensitizing stimulus. Am J Physiol Cell Physiol 2005;288(4):C769-83.