ИННОВАЦИОННЫЕ ПОДХО ДЫ К ИЗУЧЕНИЮ ЭХИНАЦЕИ
2. Г усев Е.Н. БАД -ы в натуре // Российски е аптеки.- 2000. - NQ 6. -
С.21.
3. Деримедв е дь
Л.В., Перцев И.М. , Ковалев В.Н . Биологич еск и ак
тивные д о бавки, соде ржащи е лекарственное растительно е сы р ь е // Провизор.-
2001. - NQ 3. - С. 37-40. 4. Дранник Г.М., Г риневич Ю.Я. , Дизик Г.М. И мму нотропные пре параты.- К. : Здоров'я, 1994. - 132 с. 5. КарауА о в А . В. Природные иммуностимуАяторы // Пр актикую щий врач, 1996. - NQ 1. - С. 11. 6. Котов А.Г ., Гудз енко О.П., Деркач А.Ш. Р оз ро бка н ового пре парату з iмунос ти муАюючою активнiстю на основi Echiпacea purpurea Мое псh. // Ф а р мако м, 2006.- NQ4.- С. 87-90. 7. ПиАат Т.Л., Шарманов Т.Ш., Абду,lА абенова Р.М. Ос нов ные принципы фарм аконутри циоАогии ( би ологически активные до ба вки к пище).- Астан а -Алматы-Шыкмент,
8.
Сметанина
2001. - 310 с. K.I . Фарма конут ричiологi я як наук ово
обrрунтова
ний напрямок профiлактичного викори с тання бiологiчно акт ивних
до бавок// Фарм ацев т . ж урн. -
9.
М.: Астра Ф арм Сервис,
10.
2006 . - NQ б. - С. 33-40.
Справочник ВИДАЛ Ь. Ле ка рственн ые препараты в Р осси и .
2005.- 11 68
-
с.
Тутельян В.А., Спи ричев В.Б. , Суханов Б.П . и др . Микр онутриен
ты в питании здорового и больного чеАове ка.- М . : Ко ло с,
2002.- 212
с.
11. Yip R., Daliman Р. In: Ziegel Е. and Filer L. Present knowledge in
nutrition. 7'h ed. - ILSI Press, Washington, DC. -- 1996.12. \vww. кiev. vlasti. net. 13. www. rada . net.
Р.
277-292.
PERSPECTIVES IN INVESTIGATION OF ТНЕ STRUCTURE AND PHARMACOLOGICAL EFFECTS OF POLYSACCHARIDE FRACTION FROM ECHINACEA PURPUREA (L.) MOENCH Lysiuk R.M., Rybak O.V. Danylo Halytsky Lviv National Medical Universi ty, 69, Pekarska str., 79010, Lviv, Ukraine, pharmacognosy .org.ua@ukr.net, rybak_ oksana@ukr. net
168
Раздел 11. ФИТОХИ МИЯ ЭХИНАЦЕИ ... Резюме. Обобщены и систематизированы данные о вы делении, исследовании структуры и фармакологической ак тивности полисахаридной фракции надземных и подземных органов, культуры клеток, а также индивидуальных полисаха
ридов эхинацеи пурпурной.
Summary. The article geпeralizes and systematizes the data concerning the isolation, structure and pharmaco/ogical activity of polysaccharide fraction from aboveground and suЬteгranean por tions, се// culture of purple coneflower (Е. purpurea ( L.) Moench.) and its individual polysaccharides.
Polysaccharide fractions of higher plants deserve the particular attention and are currently extensively investigated worldwide Ьу scientists, considered them the promising immunomodulating and antitumour agents. The objective of our research is summarization of data concerning isolation, chemical structure and pharmacological characters of polysaccharide fraction of various kinds of medicinal plant materials, obtained from Echinacea purpurea (L.) Moench., Asteraceae family, since the polysaccharide of the material are studied sufficiently and such investigations are carried out nowadays in several countries. Two water- soluЫe polysaccharides, PS 1 and PS 11, with immunostimulatory properties have been isolated from the aerial parts of purple coneflower (Е. purpurea). Their structures were determined as 4-0-methyl-glucuronoarablnoxylan with an average molecular weight (MW) 35,000 and an acidic arablnorhamnogalactan (MW 50,000), respectively, and they each showed significant activity in both in vitro and in vivo immunological assays [11, 17]. А pectin-like polysaccharide has been isola ted from the expressed sap and а xyloglucan (MW 79,500) from the leaves and stems of species [18]. Purple coneflower roots yie ld water-soluЬle immunostimulating polysaccharides and glycoproteins [6]. А crude polysaccharide fraction isolated from the roots of Е. purpurea appeared to have а similar coш position to that present in the aerial parts [20]. The roots of the plant contain arablnogalactans and arablnogalactan-containing glycoproteins that exert immunomodulating activity. They indicated that the glycoprot ein -containing fractions ofE. purpurea root extracts are аЬ!е to indu ce the secretion of tumor necrosis factor (TNF)-o:, interleukin (IL)-1, and INF -o: and -~ [3].
169
ИННОВАЦИОННЫЕ ПОДХОДЫ К ИЗУЧЕНИЮ ЭХИНАЦЕИ
The expense of obtaining pure polysaccharides from plant extracts and the difficulty of obtaining reproduciЬie activities led to the use of tissue culture for their isolation. The structures of the polysaccharides produced Ьу tissue culture differ from those in the aerial parts because they are primary wall components of the cultured cel\s. Thus, the polysaccharide obtained from planrs of Е. purpurea shows little similarity to that produced Ьу cell culture [7]. Three homogeneous polysaccharides, two neutral fucogalactoxyloglucans with molecular weights of 10,000 and 25,000, and an acidic arablnogalactan (MW 75,000) were isolated from cell cultures of purple coneflower. The fucogalactoxyloglucan with MW of 25,000 enhanced phagocytosis both in vitro and in vivo, while the arablnogalactan specifically stimulated macrophages to excrete the TNF [21). The supernatants from liquid cell cultшe of Е . purpurea were used to obtain highly purified arablnogalactans [9). А high degree of structural similarity between the acidic arablnorhamnogalactan from the plant material and the acidic arablnorhamnogalactan from the ceJ\ suspension was determined, using се\1 suspension studies to provide polysaccharides for immunochemical investigations. In this study, а polyclonal IgG-antibody produced from rabblts was used to study the relationship between the polysaccharide structure and Ьinding [15]. The acidic heteroxylan and fucogalactoxyloglucan were structura\ly different between the plants and the се\1 suspension cultures [11, 15]. Echinacea-derived polysaccharides are indeed active in certain immunological models [5). Two polysaccharides have been isolated from an aqueous extract of the aerial parts of Е. purpurea and shown to stimulate phagocytosis in vitro and in vivo and to enhance production of oxygen radicals Ьу macrophages in а dose-dependent way [8,17). Echinacea polysaccharides (EPS; а protein-free, blghly enriched polysaccharide mixture from the aerial parts of Е. purpurea) seem to preferentially stimulate the mononuclear immune system in vitro. EPS stimulated both peritoneal and bone marrow macrophages to behave cytotoxically in vitro. In а second experiment it was shown that EPS stimulated bone marrow macrophages to release IL-1, although it was much less potent than endotoxin iп this respect [1 ). The trial with Echinacea polysaccharides, administered Ьу injection to optimize Ьioavailabllity, demonstrated only modest results on immune 170
Раздел
11. ФИТОХИМИЯ ЭХИНАЦЕИ ...
function. In this open prospective study with matched historical controls, а polysaccharide fraction isolated from Е. purpurea herb cell cultures was tested to see if it could counter the undesired side effects of cancer chemotherapy. 15 patients with advanced gastric cancer undergoing palliative chemotherapy with а range of cytotoxic drugs also received daily iпtravenous injections of 2 mg of а polysaccharide fraction from Echinacea. While the polysaccharide treatment did appear to increase white cell counts, there were no clinically relevant effects on phagocytic activity or lymphocyte subpopulations. It has been suggested that this form of treatment should Ье investigated further [10]. ln contrast to the limited in vitro experimental evidence of immunostimulatory activity of caHeic acid derivatives and alkylamides, there is consistent and convincing evidence for the role of Echinacea polysaccharides to directly stimulate immune cells. The polysaccharide fraction increases the production of the TNF-a and the induction of inter1eukins IL-1 and IL-6 [13]. In а phase-1 clinical trial, а polysaccharide fraction, isolated from Е. purpurea tissue culture, caused an increased production of leukocytes, segmented granulocytes, and TNF-a [2]. The researchers who have worked extensively to investigate the immunostimulatory effects of polysaccharides from Echinacea [9, 14, 16, 17, 21] were successful in isolating several polysaccharide strнctures, inc luding а variety of arablnogalactans. The complex and high-molecularweight (10 to 75 kDa) polysaccharides were found to directly activate nonspecific immune cell types such as monocytes, macrophages, and natural killer (NK) cells. Echinacea polysaccharide induced stimu lation of these cell types initiated cytokine production (TNFa) and elevated phagocytic activity and oxidative burst, resulting in enhanced in vit ro and in vivo killing of Leishmania, Listeria, and Candida pathogens. lmportantly, the in vitro characteгization of the polysaccharide activity was dose dependent and with potent stimнlation occurring at concen trations 10 jlg/ ml. In addition, there is а likely mechanism of action for polysaccharide-induced stimulation of immune cell types through the Ьinding and activation of cell surface receptors present on target immune cells. The Echinacea polysaccharides were subsequently shown to activate nonspecific immune cells when evaluated in animal model s as well as lш man subjects [13, 14, 16]. This characterization of Echinacea polysaccha-
171
ИННОВАЦИОННЫЕ ПОДХОДЫ К ИЗУЧЕНИЮ ЭХИНАЦЕИ
rides is the best demonstration of in vitro bloassay activity yielding repin vivo pharmacological effects [12]. Immunostimulatory polysaccharides present in the aerial parts and produced in tissue cultures of the plant are of special interest since one of the tissue ct~lture products, an arablnogalactan, can now Ье produced on an industrial scale and is being considered for clinical trials [7]. Arablnogalactan, а highly purified polysaccharide from plant cell cultures of Echinacea purpurea, is effective in activating macrophage cytotoxicity actions against tumor cells and microorganisms (L eishmania enriettii). Тhis polysaccharide induces macrophages to produce TNF-a, IL-1, IL-6, IL-10 and interferon-~. Тhе component also induces а slight increase in T-cell proliferation [9, 13]. Macrophages, fundamentally important "helper" cells for the functional activity ofNK cells, release numerous cytokines upon stimulation with purified polysaccharides such as and including arablnogalactan, the complex carbohydrate [9, 17]. Among the resulting cytokine cascade produced Ьу such stimulated macrophages are several powerful NK enhancers, such as interferon and TNFa. Тhе study has shown that the polysaccharide, arablnogalactan, results in the production of NK stimt~lators [9]. Arablnogalactan was injected intraperitoneally into young adult and elderly inbred mice once daily for either 1 week or 2 weeks. Тhе specific arablnogalactan used is а water-soluЬle, complex carbohydra te form (L-arablno-D-galactans), а highly branched moJecule with branched backbone chains of (l-3/6)-linked ~-D-galactopyranosyl residues to which are attached side chains containing 1--arablnofuranosyl, L-arablnopyranosyl residues [ 4]. In striking contrast to these observations of increased NK cell numbers 1 week after daily administration of whoJe Echinacea [19], the results of administering arablnogalactan alone to healthy young adult mice for l week significantly decreased NK cell пumbers in the bone marrow, and resulted in no change from control nнmbers in the spleen. However, Ьу 2 weeks after daily exposure to arablnogaJactan, NK cell numbers in the bone marrow had risen to controllevels and iп the spleen they were significantly increased, almost douЬ!e the control numbers [ 4]. Тhus, unlike whole Echinacea, the effects of which were readily evident as stimulation of new NK cell production in the bone marrow Ьу 1 week [19], it appeared that 2 weeks were needed to pгoduce any stimulatory effect on NK cells when the polysaccharide alone was employed [ 4]. roduciЬle
172
Раздел 11. ФИТОХИМИЯ ЭХИНАЦЕИ ...
An acid arablnogalactan (AG) or an industrially prepared polysaccharide mixture (EPAG) tl1at differed markedly from those found in Е. purpurea, were isolated from its tissue cultures. AG induced а dose-dependent release ofTNF-a from peritoneal macrophages in vitro. When bone marrow macrophages were used in vitro, а dose-dependent release of interferon- 2а. (IL-6) was also found. The effect of Echinacea arablnogalactan from tissue cultures (AG or EPAG) is strikingly selective for macrophages in vitro [1 ]. EPAG given Ьу intravenous injection to mice at the very high dose (relative to levels in Echinacea) of 10 mg/kg resulted in а protective effect against Candida a!Ьicans infection [8]. The presented data confirm the necessity for the further investigation of polysaccharide compounds of the genus Echinacea and its related taxons. References 1. Bauer R.. Echinacea species as potential immunostimulatory drugs / R.Bauer, Н . Wagner. In: Economic and Medical Plant Research, 5'" edn./ Wagner Н., Farnsworth N. (Eds). - London: Academic Press, 1991.- Р. 253-321. 2. Beuscher N. Immonomudulierende eigenschaften von wurzelextrakten verschiedenen echinacea-arten / N.Beuscher, С. Bodinet, I. Willigmann, et al. // Z. Phytother.- 1995.- NQ 16. - Р. 157-166. 3. Bodinet С. Antiviral and immunological activity of glycoproteins from Echinacea purpurea radix / C.Bodinet, N. Beuscher // Planta Med. - 1991.- NQ 57.- Р. 33-34. 4. Currier N.L. The effect with time of administration in vivo, of the polysaccharide, arablnogalactan, on the immune and hemopoietic celllineages in murine spleen and bone marrow / N.L.Currier, D. Lejtenyi, S.C. MiJJer // Phytomedicine.- 2002.- NQ 10.- Р. 145- 153. 5. Emmendorffer А.С. Immunologically active polysaccharides from Echinacea purpurea plant and cell cultures / А.С. Emmendorffer, Н . Wagner, M.L. Lohmann-Matthes. In: Wagner Н. (Ed.). lmmunomodulatory Agents from Plants.- Basel: Birkhauser Verlag, 1999.- Р. 89-104. 6. Gruenwald J. PDR for Herbal Medicines / J.Gruenwald, Th. Brendler, Ch. Jaenicke (Sc. Eds.).- Montvale, NJ: Medical Economics Сот раnу, 2000.- 858 р. 7. HarborneJ. В. Pl1ytochemistry ofthe Genus Echinacea / J . В. Harborne, Ch.A. Williams. In: S. С. Miller (Ed.). Echinacea: the genus Echi173
ИННОВАЦИОННЫЕ ПОДХОДЫ К ИЗУЧЕНИЮ ЭХИНАЦЕИ
nacea. (Medicinal and aromatic plants - industrial profiles; v. 39). - Боса Raton, Fl.: CRC Press LLC., 2004. - Р. 66 - 82. 8. Lohmann-Matthes M.L. Aktivierund von makrophagen durch polysaccharide aus gewebedwituren von Echinacea purpurea / M.L. Lohmann-Mattl1es, Н. Wagner // Z. Phytother. - 1989.- NQ 10. - Р. 52- 59. 9. Luettig В. Macrophage activation Ьу the polysaccharide arablnogalactan isol ated from plant cell cultures ofEchinacea purpurea / В. Luettig, С. Steinmuller, G.E. Gifford, et al. // J. Natl. Cancer Inst. - 1989. NQ 81. - Р. 669-675. 10. Melchart D. Polysaccharides isolated from Echinacea purpurea herba cell cultures to counteract undesired effects of chemothera py - а pilot study / D. Melchart, С. Clemm, В. Weber et al. // Phytother. Res.2002. - NQ16.- Р. 138-142. 11. Proksch А. Structural analysis of а 4-0-methylglucuronoarablnoxylan with immunostimulating activity from Echinacea purpurea / А. P roksch, Н. Wagner / / Phytochemistry. - 1987. - NQ 26.- Р. 1989- 1993. 12. Rininger J. А. In Vitro Immunopharmacology ofEchinacea / J. А. Rininger, К. Ringer, М. Savarese. In: S. С. Miller (Ed.). Echinacea: the genus Echinacea. (Medicinal and aromatic plants-industrial profiles; v. 39). - Боса Raton, Fl.: CRC Press LLC., 2004. - Р. 173-184. 13. Roesler J . Application of purified polysaccharides from cell cultures of the plant Echinacea purpurea to test subjects mediating activation of the phagocyte system / J. Roesler, А. Emmendorffer, С. Steinmuller, et al. // Int.J. Immunopharmacol. - 1991.- NQ 13.- Р. 931- 941. 14. Roesler J. Application ofpurified polysaccharides from cell cнltu res of the plant Echinacea purpurea to mice mediates protection against systemic infections with Listeria monocytogenes and Candida alblcans / J. Roesler, С. Steinmuller, А. Kiderlen et al. // Int. J. Immunopharmacol. - 1991.- NQ 13.- Р. 27-37. 15. Schollhorn С. Immunochemical investigations of polysaccharides from Echinacea purpurea cell suspension cultures / С. Schollhom, Е. Schecklies, Н. Wagner // Planta Medica. - 1993.- NQ 59. - Р. 662- 663. 16. Steinmuller С. Polysaccharides isolated from plant cell cultures of Echinacea purpurea enhances the resistance of immunosuppressed mice against systemic infections with Candida alblcans and Listeria monocytogenes / С. Steinmuller, J. Roesler, Е. Grottrup, et al. // Int. J. Immunopharmacol. - 1993. - NQ15 . - Р. 605- 614. 174
Раздел
11. ФИТОХИМИЯ ЭХИНАЦЕИ ...
17. Stimpel М . Macrophage activation and induction of macrophage cytotoxicity Ьу purified polysaccharide fractions from the plant Echinacea purpurea 1 М. Stimpel, А. Proksch, Н. Wagner, et а!. 11 Infect. Immun.- 1984.- NQ46.- Р. 845-849. 18. Stuppner Н. Chemische und imшuno!ogiscrte lJnt ersuchungen von Polysacchariden aus der Gewebekultur von Echinacea purpurea (L.) Moench. Ph.D. thesis 1 Н . Stuppner.- lJniversity of Munich, 1985. 19. Sun L.Z.-Y. The Aшerican coneflower: а prophylactic role iпvolving non-specific iшmunity 1 L.Z.-Y. Sun, N.L. Currier, S.C. Mi!ler 11 J. Alrernative Coшplementary Med.- 1999.- NQ 5.- Р. 437-446. 20. Wagner Н. Immunstimulierend wirkende Polysaccl1aride (Heterog!ykane) aus hoheren Pflanzen 1 Н. Wagner, А. Proksch, I. Riess-Mau rer et al. 11 Arzneiш -Forsch.- 1985.- NQ 35.- Р. 1069-1075. 21. Wagner Н. Immunologically active polysaccharides of Echinacea purpurea ce!l cultures 1 H.Wagner, Н. Stuppner, W. Schafer, et а!. 11 Phytochemistry. - 1988. - NQ 27.- Р. 119-126.
ИСПОЛЬЗОВАНИЕ ЭХИНАЦЕИ ПУРПУРНОЙ ДАЯ ОБОГ АLЦЕНИЯ ДИЕТИЧЕС КИХ ПИЩЕВЫХ ПРОДУКТОВ Полякова Е.Д. " Госуниверситет- УНПК",
302020, Науrорского r . Ор е л, Россия, ivanova@ostu .ru
ш. ,
29,
Резюме. Проведены исслед ованиR минерального состава
ингредиентов пищевого обогатителR, в том чи сле эхинацеи пурпурной (соцветие, стебель, лист и корень с корневищами) четвертого года вегетации, выращенrюй в специализирован
ном хозRйстве национального парка Элементарный
состав определRЛСR
спектрального ЭДС-детектора щего микроскопа
"Орловское полесье". с помощью рентгено
miniCup JEOZ (ЯпониR).
в системе сканирую
Summary. Reseaгches of mineral composition of ingredients of the food dresser, inc/uding echinacea purple ( an inflorescence, а stalk, а lea f and а root with rhizomes) the fouгth year ofthe vegetation which has been grown up in а specialized farm of national
175
633.88+615.32 :58 42.14+53.52 и 66
УДК
БЕК
Инновационные подходы к изучению зхипацеи: Материал ы Между
И66
народной научной конференции. Полтава: Дивосвiт ,
2013. - 228
-
П олтава,
25-27
июня
2013
г.
-
с.
ISBN 978-617-633-073-8. О с вещены результаты интродукции ра:шичных видов рода Эхинацся (Ec hiпa
cea Moenclt),
особенности их биологии, культивиров ания , фитохимни и фRр:\с\ако
логи и, использования в гуманитарной медицине, зоотехнии. Изл ожены резу.~ь таты ра з работки технологий изготовления препаратов и функци о нальных про дуктов с эхинацеей.
Представленi результати iнтроду"цii рi з них видiв род у Ехiнаuея (l<; c l1 iпacea
Moench) , особливостi
iхньоi бiологil, "ультивування , фiтoxiмii
i
фар :\с\акологii , ви
користання в гуманiтарнiй медипинi, зоотехнi1. Ви"л аденнi результати роз р обки
технологiй виготовлення препаратiв та функцiональних продуктiв з ехiнапеею .
The results of different species of Echinacea genus introduction (Echinacea Moench), peculiarities of th.eir Ьiology, cultivation, photochemistry апd phaпnacology, usag·e in humaпitariaп and veterinary medicine, zootechnics are given. The results of working out the technology of preparations апd functional products with Echinacea are stated. Ergebnisse der Introduktion von verschiedenen Arten der Echinacea Gattung (Echinacea МоеnсЬ), Besordenheiten ihrer Biologie, Kul tivieren , Phitochemien und Pharmakologie, Beпнtztшg in Ншnав - 1.111d \' eteriп armediziп , ZootecJшie (Tierzucl1tlehre) sind erlaнtert. Da bei sind die Erg ebnisse der A usarbeitung der Heгste llнng stechnologoe der funktionellen Produkte aus Echiпacea dargestellt. Ре дакционная коллегия: С.В. По спелов ( ответственный редактор), В .Н . Самородов (научный редактор),
Л.А . Г.:~ущенко, И.В. Корнев, Н . Н . Опара, П . В. Пи саренко, В.Ф. Почерняева .
На первой странице обложки- новый сорт эхинацеи nурпурной
Moencl1)
(Echinacea purpшea (1.)
"Герхард Мадаус" . (Авторы В.Н. СаtVюрод ов, С. В. Поспе.Jiов). Фото С. В. Поспелова.
М а териалы поготовлены к печати Полтавским отделениеNI Укра инекого ботаничеr: кого общества, напечатаны на языке оригинала.
УДК ББК
633.88+ 615.32:58 42.14+53.52
© Полтавская государственная аграрная академия,
ISBN 978-617-633-073-8.
© Дивосвiт, 2013 .
2013.