Antifungal activity of the rhizome extracts of Pulsatilla vulgaris against Candida glabrata

  • Grażyna Łaska Department of Agri-Food Engineering and Environmental Management, Białystok University of Technology, Wiejska 45A, 15-351 Białystok, Poland
  • Aneta Sienkiewicz Department of Agri-Food Engineering and Environmental Management, Białystok University of Technology, Wiejska 45A, 15-351 Białystok, Poland
Keywords: Pulsatilla vulgaris subsp. vulgaris, Ranunculaceae, Leaves and rhizomes extracts, Biological activity, Microbiological assays


Pulsatilla vulgaris Mill. (“Pasque flower”, Ranunculaceae) is rare and a threatened plant species in Europe. It produces biologically active secondary metabolites. P. vulgaris is also known herbal drug used for centuries in traditional Chinese and Korean medicine. The rhizomes of P. vulgaris have been traditionally used for treatment of headaches, neuralgia, insomnia, hyperactivity, bacterial skin infections, septicemia, cough and bronchitis. In the present study, the extracts of leaves and rhizomes of P. vulgaris were evaluated for their antifungal, antimicrobial, antimalarial and cytotoxic activities. The results showed the antifungal activity of crude extracts of the rhizome of P. vulgaris against the yeast Candida glabrata with an IC50 of 11 µg/ml. These results indicate that the selected medicinal plant could be further investigated for identifying compounds that may be responsible for the observed activity and that may represent new leads in fungal drug discovery.



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1. Fu Y, Chen H. Studies on the pharmacognostic identification, chemical composition and pharmacology of Pulsatilla. Acta Acad Med. CPAF 2006; 5: 483-487.

2. Bang SC, Kim Y, Lee JH, Ahn BZ. Triterpenoid saponins from the roots of Pulsatilla koreana. J Nat Prod. 2005; 68: 268-272.

3. Cheon SA, Choi BK, Jeong CS, Li DW, Lee EB. The anti-inflammatory and analgesic actions of the root of Pulsatilla koreana. Korean J Pharmacogn. 2000; 31: 174-184.

4. Chung SW, Chung CG, Lim SB, Kim JK, So EH. The antimicrobial effect of Pulsatilla koreana extracts to oral micro-organism. J Korean Acad Periodontol. 2000; 30: 661-674.

5. Łaska G, Sienkiewicz A. Secondary metabolites from Pulsatilla species and their biological activity. Planta Med. 2014; 80-PN4.

6. Kim Y, Bang SC, Lee JH, Ahn BZ. Pulsatilla saponin D: the antitumor principle from Pulsatilla koreana. Arch Pharm Res. 2004; 27: 915-918.

7. Łaska G, Sienkiewicz A, Piotrowska-Niczyporuk A. Study on HPLC fingerprint characteristic of Pulsatilla patens (L.) Mill. Planta Med. 2016; S1-S381.

8. Lee SH, Byeon MS, Kim MK. Selective growth inhibitor toward human intestinal bacteria derived from Pulsatilla cernua root. J Agr Food Chem. 2001; 49: 4656-4661.

9. Cho SC, Sultan MZ, Moon SS. Anti-acne activities of pulsaquinone, hydropulsaquinone, and structurally related 1,4-quinone derivatives. Arch Pharm Res. 2009; 32: 489-494.

10. Mimaki Y, Yokosuka A, Kuroda M, Hamanaka M, Sakuma C, Sashida Y. New bisdesmosidic triterpene saponins from the roots of Pulsatilla chinensis. J Nat Prod. 2001; 64: 1226-1229.

11. Shu Z, Chen Z, Liu YL, Zhu WF, Feng YL, Xu QM, et al. A new oleanane-type triterpenoidal saponin from Pulsatilla chinensis. Nat Prod Res. 2013; 27: 2196-2201.

12. Xu K, Shu Z, Xu QM, Liu YL, Li XR, Wang YL, et al. Cytotoxic activity of Pulsatilla chinensis saponins and their structure-activity relationship. J Asian Nat Prod Res. 2013; 15: 680-686.

13. Yang H, Cho YW, Kim SH, Kim YC, Sung SH. Triterpenoidal saponins of Pulsatilla koreana roots. Phytochemistry 2010; 71: 1892-1899.

14. Xu YL, Bai L, Liu YH, Liu Y, Xu TH, Xie SH, et al. A new triterpenoid saponin from Pulsatilla cernua. Molecules 2010; 15: 1891-1897.

15. Fan WH, Liu JY, Gong YX, Ma J, Zhou N, Xu YN. A new triterpenoid saponin from Pulsatilla cernua. Nat Prod Sci. 2013; 19: 150-154.

16. Sun H, Wang Y, Zhang XQ, Zhao SX, Ye WC. Chemical constituents of Pulsatilla dahurica. Chem Nat Comp. 2009; 45: 764-765.

17. Xu HJ, Shi XW, Ji X, Du YF, Zhu H, Zhang LT. A rapid method for simultaneous determination of triterpenoid saponins in Pulsatilla turczaninovii using microwave-assisted extraction and high performance liquid chromatography-tandem mass spectrometry. Food Chem. 2012; 135: 251-258.

18. Samadder A, Das J, Das S, Khuda-Bukhsh AR. Dihydroxy-isosteviol-methyl-ester, an active biological component of Pulsatilla nigricans, reduces arsenic induced cellular dysfunction in testis of male mice. Environ Toxicol Phar. 2012; 34: 743-752.

19. Rolski S, Przyborowski L. Dissert Pharm. 1961; 13: 349-355.

20. Ye WC, Ji NN, Zhao SX, Che CT. A New Cytotoxic Saponin from Pulsatilla patens var. multifida. Pharm Biol. 2001; 39: 7-10.

21. Danova K, Bertoli A, Pistelli L, Dimitrow D, Pistelli L. In vitro culture of Balkan endemic and rare Pulsatilla species for conservational purposes and secondary metabolites production. Bot Serb. 2009; 33: 157-162.

22. Sharma V, Łaska G, Radhakrishnan SVS, Jacob MR, Zjawiony JK. Phytochemical investigation and pharmacological evaluation of Pulsatilla patens var. patens. Planta Med. 2014; 80-PD3.

23. Łaska G, Sienkiewicz A, Piotrowska-Niczyporuk A. New Polish species with isolated Pulsatilla saponin D. Planta Med. 2016; 82: S1-S381.

24. Luft G. The distribution of saponins and tannins in the plant. Monatshefte fuer Chemie 1926; 47: 259-284.

25. Raymond-Hamet L. Pharmacodynamic properties of Anemone pulsatilla. Bull Sci Pharmacol. 1927; 34: 143-151.

26. Kroeber L. On the pharmacology and therapeutic use of the anemonine drugs from Ranunculaceous plants. Pharmazie. 1949; 4: 181-190.

27. Weryszko-Chmielewska E, Sulborska A. Staminodial nectary structure in two Pulsatilla (L.) species. Acta Biol Cracov Ser Bot. 2011; 53: 94-103.

28. Liu DD, Chao WM, Turgeon R. Transport of sucrose, not hexose, in the phloem. J Exp Bot. 2012; 63: 4315-4320.

29. Hensen I, Oberprieler C, Wesche K. Genetic structure, population size, and seed production of Pulsatilla vulgaris Mill. (Ranunculaceae) in Central Germany. Flora 2005; 200: 3-14.

30. DiLeo MF, Graf R, Holderegger R, Rico Y, Wagner HH. Highly polymorphic microsatellite markers in Pulsatilla vulgaris (Ranunculaceae) using next-generation sequencing. Appl Plant Sci. 2015; 3(7): apps.1500031.

31. Walker KJ, Pinches CE. Reduced grazing and the decline of Pulsatilla vulgaris Mill. (Ranunculaceae) in England. UK. Biol Conserv. 2011; 144: 3098-3105.

32. Piqueray J, Saad L, Bizoux JP, Mahy G. Why some species cannot colonise restored habitats? The effects of seed and microsite availability. J Nat Conserv. 2013; 21: 189-197.

33. Pfeifer E, Holderegger R, Matthies D, Rutishauser R. Investigation on the population biology of a flagship species of dry meadows: Pulsatilla vulgaris Mill. in north-eastern Switzerland. Bot Helv. 2002; 112: 153-172.

34., 2000-2014. know more be sure, Auckland 0632 New Zealand: Mission Statement “to empower patients with the knowledge to better manage their own healthcare and to improve consumer safety by assisting in the reduction of medication errors”.

35. Bundesministerium für Gesundheit, für alle. (accessed 25 February 2014).

36. Bone K. A clinical guide to blending liquid herbs: herbal formulations for the individual patient. Churchill Livingstone, St Louis, Missouri, 2003.

37. Kanetoshi A, Inoue S, Anetai M, Fujimoto T, Aoyagi M, Sato M. An in vitro screening test for anti-cancer components of wild plants in Hokkaido. Report-Hokkaido Institute of Public Health. 2005; 55: 49-53.

38. Łaska G, Sienkiewicz A, Stocki M, Sharma V, Zjawiony JK, Jacob M. Secondary metabolites from Pulsatilla patens and Pulsatilla vulgaris and their biological activity. Planta Med. 2015; 81-PM_122.

39. Saify ZS, Noor F, Mushtaq N, Dar A. Assessment of Anemone pulsatilla for some biological activities. Pak J Pharm Sci. 1998; 11: 47-53.

40. Gasnier C, Laurant C, Decroix-Laporte C, Mesnage R, Clair E, Travert C, et al. Defined plant extracts can protect human cells against combined xenobiotic effects. J Occup Med Toxicol. 2011; 6: 3.

41. Jürgens A, Dötterl S. Chemical composition of pollen volatiles in Ranunculaceae: Genera-specific profiles in Anemone, Aquilegia, Caltha, Pulsatilla, Ranunculus, and Trollius species. Am J Bot. 2004; 91: 1969-1980.

42. Mills S, Bone K. The essential guide to herbal safety. Elsevier, St Louis, 2005.

43. Foster S, Johnson R. National geographic desk reference to natural medicine. National Geographic, Washington, 2006.

44. Baer H, Holden M, Seegal BC. The nature of the antibacterial agent from Anemone pulsatilla. J Biol Chem. 1946; 162: 65-68.

45. Duan H, Zhang Y, Xu J, Qiao J, Suo Z, Hu G, et al. Effect of anemonin on NO, ET-1 and ICAM-1 production in rat intestinal microvascular endothelial cells. J Ethnopharmacol. 2006; 104: 362-366.

46. Bang SC, Lee JH, Song G, Kim DH, Yoon MT, Ahn BZ. Antitumor activity of Pulsatilla koreana saponins and their structure-activity relationship. Chem Pharm Bull. 2005; 53: 1451-1454.

47. NCCLS: National Committee on Clinical Laboratory Standards, 2002; 22: 15.

48. NCCLS: National Committee on Clinical Laboratory Standards, 2002; 22: 16.

49. NCCLS: National Committee on Clinical Laboratory Standards, 2003; 23: 18.

50. NCCLS: National Committee on Clinical Laboratory Standards, 2006; 26: 2.

51. Franzblau SG, Witzig RS, McLaughlin JC, Torres P, Madico G, Hernandez A, et al. Rapid, low-technology MIC determination with clinical Mycobacterium tuberculosis isolates by using the microplate Alamar blue assay. J Clin Microbiol. 1998; 36: 362-366.

52. Makler MT, Hinrichs DJ. Measurement of the lactate dehydrogenase activity of Plasmodium falciparum as an assessment of parasitemia. Am J Trop Med Hyg. 1993; 48: 205-210.

53. Borenfreund E, Babich H, Martin-Alguacil N. Rapid chemosensitivity assay with human normal and tumor cells in vitro. In Vitro Cell Dev Biol. 1990; 26: 1030-1034.

54. Xu QM, Shu Z, He WJ, Chen LY, Yang SL, Yang G, et al. Antitumor activity of Pulsatilla chinensis (Bunge) Regel saponins in human liver tumor 7402 cells in vitro and in vivo. Phytomedicine 2012; 19: 293-300.

55. Hong SW, Jung KH, Lee H, Son MK, Yan HH, Kang NS, et al. SB365, Pulsatilla saponin D, targets c-Met and exerts antiangiogenic and antitumor activities. Carcinogenesis 2013; 34: 2156-2169.

56. Han CK, Park YH, Jin DQ, Hwang YK, Oh KB, Hand JS. SK-PC-B70M from Pulsatilla koreana improves scopolamine-induced impairments of memory consolidation and spatial working memory. Brain Res. 2007; 1184: 254-259.

57. Seo JS, Kim TK, Leem YH, Lee KW, Park SK, Baek IS, et al. SK-PC-B70M confers anti-oxidant activity and reduces β levels in the brain of Tg2576 mice. Brain Res. 2009; 1261: 100-108.

58. Yoo HH, Lee SK, Lim SY, Kim Y, Kang MJ, Kim EJ, et al. LC-MS/MS method for determination of hederacolchiside E, a neuroactive saponin from Pulsatilla koreana extract in rat plasma for pharmacokinetic study. J Pharm Biomed Anal. 2008; 48: 1425-1429.

59. Liu JY, Guan YL, Zou LB, Gong YX, Hua HM, Xu YN, et al. Saponins with neuroprotective effects from the roots of Pulsatilla cernua. Molecules 2012; 17: 5520-5531.

60. Dai L, Wang H, Chen Y. The immune enhancing effect of PcG2A-a glycoprotein isolated from dried root of Pulsatilla chinensis (Bunge) Regel. Chin J Biochem Pharm. 2000; 21: 230-231.

61. Li HB, Wong CC, Cheng KW, Chen F. Antioxidant properties in vitro and total phenolic contents in methanol extracts from medicinal plants. Food Sci Tech. 2008; 41: 385-390.

62. Son MK, Jung KH, Lee HS, Lee H, Kim SJ, Yan HH, et al. SB365, Pulsatilla saponin D suppresses proliferation and induces apoptosis of pancreatic cancer cells. Oncol Rep. 2013; 30: 801-808.

63. Fidel F, Vazquez J, Sobel J. Candida glabrata: review of epidemiology, pathogenesis, and clinical disease with comparison to C. albicans. Clin Microbiol Rev. 1999; 12: 80-96.

64. Bethea EK, Carver BJ, Montedonico AE, Reynolds TB. The inositol regulon controls viability in Candida glabrata. Microbiology 2009; 156: 452-462.

65. Details/ (accessed 11 October 2012).

66. Details/ (accessed 12 June 2008).

67. Details/ (accessed 4 October 2005).

68. Böcher TW. Beiträge zur Zytologie der Gattung Anemone. Botanisk Tidsskrift 1934; 42: 183-206.

69. Butaye J, Adriaens D, Honnay O. Conservation and restoration of calcareous grasslands: A concise review of the effects of fragmentation and management on plant species. Biotechnol Agro Soc Environ. 2005: 9: 111-118.

70. IUCN, The IUCN Red List of threatened species. Version 2014.1. (accessed 12 June 2014).

71. Witkowski ZJ, Król W, Solarz W, eds. Carpathian list of endangered species. WWF and Institute of Nature Conservation, Polish Academy of Sciences, Vienna-Krakow, 2003.

72. Gärdenfors U. The 2010 Red List of Swedish species. ArtDatabanken, SLU, Uppsala, 2010.

73. Cheffings C, Farrell L, eds. The vascular plant red data list for Great Britain. Joint Nature Conservation Committee, Peterborough, 2005.

74. Ludwig G, Schnittler M. Red List of threatened plants in Germany. Bundesamt für Naturschutz, Bonn, 1996.

75. Moser D, Gygax A, Bäumler B, Wyler N, Palese R. Red List of the threatened ferns and flowering plants of Switzerland. Bundesamt für Umwelt, Wald und Landschaft, Zentrum des Datenverbundnetzes der Schweizer Flora, Conservatoire et Jardin botaniques de la Ville de Genève, Bern, Chambésy, 2002.

76. Niklfeld H, Schratt-Ehrendorfer L, eds. Red List of threatened plants of Austria. Grüne Reihe, Bundesministerium für Umwelt, Jugend und Familie, Vienna, 1999.

77. Zarzycki K, Szeląg Z. 2006. Red list of vascular plants in Poland. In: Mirek Z, Zarzycki K, Wojewoda W, Szeląg Z, eds. PAN, Inst. Bot., Kraków, 9-20.

78. Kaźmierczakowa R, Zarzycki K, Mirek Z. Polish Red Data Book of Plants. PAN, Inst. Bot., Kraków, 2014.

79. Franz E. Population development, habitat preference and causes of endangerment of the Pasque Flower (Pulsatilla vulgaris Mill.) in Austria between 1991 and 2005. Linzer Biologische Beitraege 2005; 37: 1145-1176.

80. Colling G. Red list of the vascular plants of Luxembourg. Ferrantia 2005; 42: 1-77.

81. Rassi P, Alanen A, Kanerva T, Mannerkoski I, eds. The 2000 Red List of Finnish species. Ympäristöministeriö & Suomen ympäristökeskus, Helsinki, 2001.
How to Cite
Łaska, G.; Sienkiewicz, A. Antifungal Activity of the Rhizome Extracts of Pulsatilla Vulgaris Against Candida Glabrata. European Journal of Biological Research 2019, 9, 93-103.
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