www.elsevier.com/locate/fitote

Fitoterapia 75 (2004) 577–580

Short report

Antimicrobial activity and cytotoxicity of

Cirsium tenoreanum

Monica Rosa Loizzo a,*, Giancarlo A. Statti a, Rosa Tundis a,Filomena Conforti a, Sebastiano Ando’ b, Francesco Menichini a

aDipartimento di Scienze Farmaceutiche, Universita della Calabria, 87036 Rende (CS), ItalybDipartimento di Biologia Cellulare, Facolta’ di Farmacia, Universita della Calabria,

87036 Rende (CS), Italy

Received 8 July 2003; accepted 16 March 2004

Available online 28 July 2004

Abstract

The methanol and ethyl acetate extracts of the aerial parts of Cirsium tenoreanum, have been

evaluated for their phytochemical constituents and antibacterial and cytotoxic properties. Three

flavonoids, apigenin, quercetin-3-O-galactoside and kaempferol-3-O-ramnoside, were isolated. The

ethyl acetate extract showed a good antiproliferative activity.

D 2004 Elsevier B.V. All rights reserved.

Keywords: Cirsium tenoreanum Petrak; Antibacterial activity; Cytotoxicity

Plant. Cirsium tenoreanum Petrak [1], aerial parts collected in the flowering season in

Pollino Mountain, Calabria, and identified by Prof. G. Cesca, Botanical Garden,

University of Calabria, Italy. A voucher specimen is deposited in the Herbarium of

the Botanical Garden.

Uses in traditional medicine. C. tenoreanum is used for the treatment of varicose

veins [2].

0367-326X/$ - see front matter D 2004 Elsevier B.V. All rights reserved.

doi:10.1016/j.fitote.2004.03.011

* Corresponding author. Tel.: +39-0-984-493063; fax: +39-0-984-493298.

E-mail address: mr.loizzo@unical.it (M.R. Loizzo).

Table 1

MIC of C. tenoreanum ethyl acetate extract*

Microorganism EtOAc extract

S. aureus subsp. aureus 0.5

S. aureus MRSA 0.5

B. subtilis N.D.

E. coli HB101 1

E. coli pcDNA3 N.D.

Methanol extract: no detected activity.* (mg/ml).

M.R. Loizzo et al. / Fitoterapia 75 (2004) 577–580578

Previously isolated constituents. Flavonoids, acids [3–5].

Tested material. Methanol (yield: 7.81%) and ethyl acetate (yield: 0.26%) extracts.

Studied activity. Antibacterial activity by determination of the minimum inhibitory

concentration (MIC) using the tube dilution method [6] and the h-lactamase inhibition

activity by plate bioassay. Antiestrogenic activity by Trypan blue dye exclusion assay

using Wild-type human breast cancer MCF-7 cells (MCF7-wt) [7].

Used microorganisms. Microorganism listed in Tables 1 and 2 were collected from

Centro Sanitario, University of Calabria and Oxoid, Italy.

Cells. MCF-7 wt cells was routinely cultured in two different conditions of growth:

DMEM low glucose lacking red phenol and DMEM with red phenol, both supplemented

with 10% non-essential aminoacids, 10% FCS, 1% penicillin/streptomicin and 10% of L-

glutamine, in a humidified atmosphere with 95% O2, 5% CO2 at 37 jC; pH of the media

was monitored at 7.4.

Results. Summarized in Tables 1–4.

Table 2

h-Lactamase inhibition assay of C. tenoreanum methanolic and ethyl acetate extracts

Microorganism MeOH extract EtOAc extract

Concentration (mg/ml) IZD (mm) Concentration (mg/ml) IZD (mm)

E. coli pcDNA3 1 50 MIC/2 50

0.5 35 MIC/4 37

0.25 25 MIC/8 20

MIC/16 –

IZD: inhibition zones diameter; – : did not show any inhibitory activity; Control: Ampicillin 5 Ag/ml.

Table 3

Effect of C. tenoreanum methanolic and ethyl acetate extracts on MCF7-wt cell line growth in DMEM

Extracts Concentration (Ag/ml) % of cell death

24 h 48 h 72 h

MeOH

DMSO 0.5% (v/v) 0 0 0

5 19.04 12.5 20

10 19.05 27.8 40.2

50 23.80 33.3 40.5

100 42.8 60 61

EtOAc

DMSO 0.5% (v/v) 0 0 0

5 6.7 5.8 6.2

10 26.7 17.6 12.5

50 33.3 17.7 25

100 37.7 41.2 46

Table 4

Effect of C. tenoreanum methanolic and ethyl acetate extracts on MCF7-wt cell line growth in DMEM low

glucose

Extracts Concentration (Ag/ml) % of cell death

24 h 48 h 72 h

MeOH

DMSO 0.5% (v/v) 0 0 0

5 9.6 8.3 20.2

10 18.2 30.5 40.6

50 18.4 33.5 41.12

100 45.4 62 63.8

EtOAc

DMSO 0.5% (v/v) 0 0 0

5 20.25 28.5 29.5

10 40.2 35.7 31.7

50 40.4 37.6 52

100 70.7 71.4 73

M.R. Loizzo et al. / Fitoterapia 75 (2004) 577–580 579

Conclusions. The results indicate that ethyl acetate extract possesses a mild antimicrobial

activity. Both extracts showed h-lactamase inhibition and possess antiproliferative

activities, probably due to the presence of flavonoids.

Acknowledgements

The authors are grateful to Prof. G. Cesca of the Botany Department at University of

Calabria, Italy, for supplying the herb samples used in this study and Prof. F. Delle

M.R. Loizzo et al. / Fitoterapia 75 (2004) 577–580580

Monache of the Istituto di Chimica e Clinica Chimica, Universita Sacro Cuore, Rome,

Italy, for spectroscopic analysis.

References

[1] Pignatti S. Flora d’Italia. Bologna: Edagricole; 1982.

[2] Hafliger E, Brun-Hol J. Tavola delle Malerbe. Basle, Switzerland: CIBA-GEIGY Edizioni; 1981.

[3] Binder RG, Benson M, Haddon WF, French RC. Phytochemistry 1992;31:1033.

[4] Park JC, Lee JH, Choi JS. Phytochemistry 1995;39:261.

[5] Statti G, Menichini F, Delle Monache F. Fitoterapia 1997;68:402.

[6] Sham DF, Washington JA II. Antibacterial susceptibility test: diluition metods. In: Balows A, Hausler WJ,

Herrmann KL, Isenbrg HD, Shadomy HJ, editors. 5th ed. Manual of clinical microbiology. Washington DC

American Society for Microbiology; 1991.

[7] Moldens P, Hogberg J, Overhenius S. In: Flisher S, Packer L, editors. Methods in enzymology. New York:

Academic Press; 1978.