Journal of Herbs, Spices & Medicinal Plants

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Ethnomedical Survey and Cytotoxic Activity of Medicinal Plant Extracts Used in Kohgiluyeh and Boyerahmad Province in Iran Mahmoud Mosaddegh b

a b

, Somayeh Esmaeili

, Maryam Hamzeloo Moghadam

Hamid Moazzeni

a c

a c

, Ali Haeri

, Farzaneh Naghibi

a d

a

a

, Atefeh Pirani &

a

a

Traditional Medicine and Materia Medica Research Center , Shahid Beheshti University of Medical Sciences , Tehran , Iran b

School of Pharmacy , Shahid Beheshti University of Medical Sciences , Tehran , Iran c

School of Traditional Medicine , Shahid Beheshti University of Medical Sciences , Tehran , Iran d

School of Medicine , Shahid Beheshti University of Medical Sciences , Tehran , Iran Published online: 21 Jun 2012.

To cite this article: Mahmoud Mosaddegh , Somayeh Esmaeili , Farzaneh Naghibi , Maryam Hamzeloo Moghadam , Ali Haeri , Atefeh Pirani & Hamid Moazzeni (2012) Ethnomedical Survey and Cytotoxic Activity of Medicinal Plant Extracts Used in Kohgiluyeh and Boyerahmad Province in Iran, Journal of Herbs, Spices & Medicinal Plants, 18:3, 211-221, DOI: 10.1080/10496475.2012.671801 To link to this article: http://dx.doi.org/10.1080/10496475.2012.671801

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Journal of Herbs, Spices & Medicinal Plants, 18:211–221, 2012 Copyright © Taylor & Francis Group, LLC ISSN: 1049-6475 print/1540-3580 online DOI: 10.1080/10496475.2012.671801


Ethnomedical Survey and Cytotoxic Activity of Medicinal Plant Extracts Used in Kohgiluyeh and Boyerahmad Province in Iran MAHMOUD MOSADDEGH,1,2 SOMAYEH ESMAEILI,1,3 FARZANEH NAGHIBI,1,2 MARYAM HAMZELOO MOGHADAM,1,3 ALI HAERI,1,4 ATEFEH PIRANI,1 and HAMID MOAZZENI1 1

Traditional Medicine and Materia Medica Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2 School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran 3 School of Traditional Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran 4 School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

Methanol extracts of selected medicinal species used in Kohgiluyeh and Boyerahmad (Iran) were evaluated in vitro for their cytotoxic activity against MCF7, HepG2, MDBK, and A549 cells through MTT assay. The extract from Dorema aucheri demonstrated cytotoxicity with IC50 of 20.09 and 48.65µg.mL−1 against HepG2 and A549 cells, respectively, which was further fractionated and subjected to MTT assay. The results may support the non-cytotoxicity of most plant species used traditionally as natural remedies. KEYWORDS Ethnobotany, ethnopharmacology, MTT assay

INTRODUCTION The potential of bioactive plants or their extracts to provide new and novel products for disease treatment and prevention is enormous (35). Indigenous Received December 11, 2011. The authors thank the contribution of the local healers of Kohgiluyeh and Boyerahmad, Iran, especially Miss Azar Ravangard, who participated in interviewing about natural remedies of the province. Address correspondence to Maryam Hamzeloo Moghadam, Department of Traditional Pharmacy, School of Traditional Medicine, Shahid Beheshti University of Medical Sciences, 8 Shams Alley, Vali-e-Asr Street, Tehran 1516745811, Iran. E-mail: [email protected] 211

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populations use a wide range of plants to maintain their health, and there is great promise for new drug discovery based on traditional plant uses for therapeutic purposes (10). Kohgiluyeh and Boyerahmad is a mountainous region located in southwest Iran, with a population of 634,299, according to the 2006 census (32). Despite the diversity and great potential of the medicinal herbs of this province, little scientific work has been done. Traditionally, many of the species have been used as infusions, deccotions, or other forms of preparations to treat illnesses but have not been investigated sufficiently for safety and cytotoxicity. This study evaluated the antiproliferative effects of 16 plant extracts and 3 fractions of species selected based on information collected by interviewing local healers of the province (Table 1).

MATERIAL AND METHODS Plant Material Sixteen plant species were collected from Kohgiluyeh and Boyerahmad province, Iran (2009–2010) and were authenticated by botanists at the Traditional Medicine and Materia Medica Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran. All voucher specimens were deposited at the Traditional Medicine and Materia Medica Research Center herbarium for future reference. The local names and traditional use, time of collection, mode of administration, part of the plants listed as being used, and parts were collected and subjected to the cytotoxicity assay are presented (Tables 1 and 2).

Extraction Dried powdered plants (10 g) were macerated with MeOH at room temperature for 24 h. Where the upper part of the plant (flowers or leaves) was reported as being used, the entire aerial part including flowers and leaves were extracted. In some cases, as in Gentiana olivieri and Haussknechtia elymaitica, although only the underground parts were used traditionally, both underground and aerial parts were extracted separately and examined. The extracts were dried using a rotary evaporator at temperature not exceeding 40◦ C and tested for cytotoxicity.

Fractionation Dorema aucheri that demonstrated cytotoxic activity was selected for fractionation. Then 30 g of dried powdered plant was macerated with petroleum ether at room temperature; after 24 h, it was filtered, and the

213

Daphne mucronata

Descurainia sophia

Dorema aucheri

Gentiana olivieri

4

5

6

Arctium minus

2

3

Achillea wilhelmsii

Species

1

No.

Gentianaceae

Apiaceae

Brassicaceae

Thymelaceae

Asteraceae

Asteraceae

Family

Malandar /malandar/

Bilhar / bilhar/

Khak-shir /k6k ir/

Kheshk / keek/

Babaadam /b6b6 6dam/

Berenjas / berendZ6s/

Local name

late spring, summer

spring, summer

spring

spring

Time of traditional collection∗

late winter, summer for disinfecting wounds/ spring externally: powdered drug

bleeding, high blood sugar and pressure/ infusion rheumatoid complaints, kidney stones (leaves), skin diseases (roots)/ infusion joint pain/ ointment made from concentrated decoction stomach disorders, constipation/ macerate constipation/ cooked

Traditional use / mode of preparation∗

aerial parts, underground part

root

(Continued)

tuber

aerial parts

aerial parts

aerial parts, Tuberous roots

aerial parts

Parts extracted in this study

aerial parts

seeds

stem bark

leaves and root

flowers

Parts Traditionally used∗

TABLE 1 Data of Plant Species and Extractions Used for Medicinal Purposes by the Populations in Kohgiluyeh and Boyerahmad Province, Iran and the Parts Used for Extraction and Cytotoxicity Examination in the Study


214

∗ Unpublished

13

12

11

data.

Scrophularia striata Stachys pilifera

Lepidium draba Lonicera nummulariifolia Salvia sclarea

9 10

8

Haussknechtia elymaitica Helichrysum oligocephalum

Species

7

No.

TABLE 1 (Continued)

Lamiaceae

Scrophulariaceae

Lamiaceae

Brassicaceae Caprifoliaceae

Asteraceae

Apiaceae

Family

Bonje-mari/ bondZem6ri/ O-lile /oli le/

Marvareshk / marvare k/

Sozah /seu zah/ Shen / en/

Darameye karimkhani / darameye karim k6ni/

Kelos/ke leus/

Local name high blood sugar and pressure/ cooked cold (in combination with other herbs)/ powder in combination with other herbs fever, colds/ infusion esophagus wounds/ decoction colds, dry cough/ cooked in combination with other herbs menorrhagia after child birth /inhalation menstrual disorders, in particular amenorrhea/ decoction

Traditional use / mode of preparation∗

spring

spring

spring

aerial parts

aerial parts

aerial parts

aerial parts leaves

aerial parts

spring

spring spring

aerial parts

Parts Traditionally used∗

spring, summer

Time of traditional collection∗


aerial parts

aerial parts

whole plant

aerial parts whole plant

basal leaves, Tuber whole plant

Parts extracted in this study

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Achillea wilhelmsii

Arctium minus Daphne mucronata

Descurainia sophia

Dorema aucheri

2 3

4

5

Species

1

No.

Analgesic, anti-inflammatory (19)

Anti-oxidant (31) stimulant of humoral and cellular immune functions in mice (30) total cholesterol and LDL-cholesterol, triglyceride lowering effect decrease was observed in diastolic and systolic blood pressure after 2 and 6 months, respectively (5). Antihypertensive activity (8) Gnidilatimonoein (diterpene): cytotoxic (39) Gnidilatimonoein (diterpene): apoptotic (17) Relieve cough, prevent asthma, reduce edema, promote urination, cardiotonic effect (33)

Effects

(Continued)

cis-beta-ocimene, menthol, neoisomenthyl acetate, alloaromadendrene, longicyclene (16),descurainoside, sinapic acid (33), descorainolide A,B, descurainin, strophantidin, isorhamnetin, descuraic acid (34), descurainin A and descurainoside B, quercetin-3-O-beta-D-glucopyranosyl-7-O-beta-gentiobioside, kaempferol-3-O-beta-D-glucopyranosyl-7-O-beta-gentiobioside, isorhamnetin-3-O-beta-D-glucopyranosyl-7-O-beta-gentiobioside, quercetin-7-O-beta-gentiobioside, kaempferol-7-O-beta-gentiobioside, isorhamnetin-7-O-beta-gentiobioside, quercetin-3,7-di-O-beta-D-glucopyranoside, kaempferol-3, 7-di-O-beta-D-glucopyranoside, isorhamnetin-3, 7-di-O-beta-D-glucopyranoside, kaempferol-3-O-beta-Dglucopyranosyl-7-O-[(2-O-trans-sinnapoyl)-beta-Dglucopyranosyl(1–>6)]-beta-D-glucopyranoside), sinapic acid ethyl ester and 3, 4, 5-trimethoxyl-cinnamic acid (36) Flavonoids (7,19), Coumarines (7)

peptides (8) Daphnecin, aquillochin, umbelliferone, and coumarin (27) gnidilatimonoein (39)

2-methyl butanal, alpha-pinene, alpha-thujene, camphene, hexanal, beta-pinene and 1,8-cineol (9), camphor, borneol, linalool, 1,8-cineole, chrysanthenol acetate, and carvacrol (1)

Previously isolated compounds

TABLE 2 Previously Published Data on the Therapeutic Properties and Chemical Compounds Isolated from The Plant Species Used by the Populations in Kohgiluyeh and Boyerahmad Province, Iran


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Haussknechtia elymaitica Helichrysum oligocephalum Lepidium draba Lonicera nummulariifolia Salvia sclarea

7

Scrophularia striata

Stachys pilifera

12

13

11

9 10

8

Gentiana olivieri

Species

6

No.

TABLE 2 (Continued)

—

Anti-inflammatory (6) matrix metalloproteinases inhibitor (11)

Antibacterial (15,36) anti-inflammatory, analgesic (essential oil) (24) Antidepressant (28) essential oil: cytotoxic (14)

— —

—

Hypoglycaemic, antihyperlipidemic (29), hepatoprotective (2,23) Immunomodulatory effect (4)

Effects

Scareol, manool, salvipisone, ferruginol, microstegiol, candidisiol, 7-oxoroyleanone, 2,3-dehydrosalvipisone, 7-oxoferruginol-18-al, as well as two sesquiterpenes, caryophyllene oxide and spathulenol, alpha-amyrin, beta-sitosterol, apigenin, luteolin, 4’-methylapigenin, 6-hydroxyluteolin-6,7,3’,4’-tetramethyl ether, 6-hydroxy apigenin-7,4’-dimethyl ether (13,37,38) essential oil: beta-myrcene, linalool, linalyl acetate, linalyl formate, trans-caryophyllen, alpha-terpineol, geranyl formate, germacrene, neryl acetate, geranyl acetate, neryl alcohol, geraniol, caryophyllene oxide, spathulenol (12), 1-oxoaethiopinone (13) cinnamic acid, three flavonoids (quercetine, isorhamnetin-3-O-rutinoside and nepitrin) and one phenyl propanoid glycoside (acteoside 1) (20) trans- verbenol, cis- chrysanthenyl acetate (18)

glucoraphanin (25) —

—

—

Isoorientin (23)

Previously isolated compounds


Ethnomedical Survey and Cytotoxic Activity

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filtrate was concentrated, the residue of the plant was treated with chloroform for another 24 h for the chloroform fraction to be prepared (CF), and the steps were repeated for methanol fraction. The concentrated fractions were then subjected to the cytotoxicity assay.

Preparation of Extracts and Fractions for MTT Assay


The extracts and fractions were dissolved in dimethyl sulfoxide (DMSO): 10 mg.mL−1 of each sample to make stock solutions. Serial dilutions were prepared accordingly from the above stock solution to get final concentrations with DMSO not exceeding 1%.

Cell Lines MCF7 (human breast adenocarcinoma), HepG2 (hepatocellular carcinoma), MDBK (bovine kidney cells), and A549 (non-small-cell line carcinoma) cells were obtained from Pasteur Institute, Tehran, Iran. MCF7 cells were cultured in Dulbecco’s modified eagle medium (DMEM; Gibco) with 5% fetal bovine serum (FBS; Gibco) while the other three cell lines were cultured in RPMI 1640 medium (Sigma) with 10% FBS to maintain the desired growth. All cell lines were treated with 1% penicillin-streptomycin (Sigma) in a humidified incubator at 37◦ C in an atmosphere of 5% CO2 . The growth curve of each cell line was assessed.

MTT Assay Cell viability was assessed in a micro-culture tetrazolium/formazan assay (MTT assay), as given earlier (3) with some modifications (21,22,26) in the absence and presence of different concentrations of the compounds. The cells were seeded in 96-well plates at 11 × 103 for MDBK cells, 9 × 103 for MCF7, 15 × 103 for HepG2, and 8 × 103 for A549 cells and incubated at 37◦ C. After 24 h, the medium was replaced with fresh medium containing concentrations of compounds to be tested at (in µg.mL−1 ) 100, 50, 25, 12.5, 6.25, and 3.125. After 72-h exposure of cells at 37◦ C to each extract, the medium was replaced with fresh medium containing MTT ([3-(4,5-dimethylthiazol-2yl)-2,4-diphenyltetrazolium bromide]; Sigma), with a final concentration of 0.5 mg.mL−1 . The cells were incubated for another 4 h in a humidified atmosphere at 37◦ C, then the medium containing MTT was removed, and the remaining formazan crystals were dissolved in DMSO. The absorbance was recorded at 570 nm with an enzyme-linked immunosorbent assay reader (TECAN), using Tamoxifen (22) as positive control. The relative cell viability (%) related to control wells was calculated by [A]samples / [A]control × 100 where [A]samples is the absorbance of test sample and

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[A]control is the absorbance of wells containing cells, cell culture medium, and DMSO 1%. The dose-response curves were graphed by the Microsoft Excel program, and IC50 values were assessed.


RESULTS Nineteen samples (16 plant extracts and 3 fractions of Dorema aucheri) were evaluated for cytotoxic activity. Extracts with IC50 > 100 µg.mL−1 in MTT assay were considered inactive (all but D. aucheri extract). In vitro bioassay evaluation demonstrated that D. aucheri MeOH extract exhibited cytotoxic activity in HepG2 (IC50 = 20.09 µg.mL−1 ) and A549 cells (IC50 = 48.65 µg.mL−1 ); fractions of D. aucheri (petroleum ether fraction and chloroform fraction) showed cytotoxic activity in all four cell lines while fractionation with methanol had no cytotoxic activity against any of the cell lines (Figure 1).

DISCUSSION Of all the plant species examined, D. aucheri tuber extract and fractions demonstrated cytotoxic activity. Although cytotoxic diterpenes have been reported from Daphne mucronata (17,39), its extract showed no cytotoxic activity with the concentrations tested for MTT assay in this study. More investigations with higher concentrations of the extract might show cytotoxic activity. The essential oil of Salvia sclarea had been reported to have cytotoxic activity (14) but the extract was not cytotoxic in this study. This

FIGURE 1 The IC50 values of Dorema aucheri methanol extract and fractions in petroleum ether (PF), chloroform (CF), and methanol (MF) against MCF7, HepG2, MDBK and A549 cells.


219

study identified non-cytotoxicity of a majority of plant extracts used in traditional healing in the Kohgiluyeh and Boyerahmad province in Iran and suggested groundwork for evaluating the biologic activity of the traditional use of plant species.


REFERENCES 1. Afsharypuor, S., S. Asgary, and G. B. Lockwood. 1996. Constituents of the essential oil of Achillea wilhelmsii from Iran. Planta Med. 62(1):77–78. 2. Aktay, G., D. Deliorman, E. Ergun, F. Ergun, E. Ye¸silada, and C. Cevik. 2000. Hepatoprotective effects of Turkish folk remedies on experimental liver injury. J. Ethnopharmacol. 73(1–2):121–129. 3. Alley, M. C., D. A. Scudiero, A. Monkes, M. L. Hursey, M. J. Czerwinski, D. L. Fine, B .J. Abbott, J. G. Mayo, R. H. Shoemaker, and Boyd M.R . 1988. Feasibility of drug screening with panel of human tumor cell lines using a microculture tetrazolium assay. Cancer Res. 48:589–601. 4. Amirghofran, Z., A. Azadmehr, and K. Javidnia. 2007. Haussknechtia elymatica: A plant with immunomodulatory effects. Iranian J. Immunol. 4(1):26–31. 5. Asgary, S., G. H. Naderi, N. Sarrafzadegan, N. Mohammadifard, S. Mostafavi, and R. Vakili. 2000. Antihypertensive and antihyperlipidemic effects of Achillea wilhelmsii. Drugs Exp. Clin. Res. 26(3):89–93. 6. Azadmehr, A., A. Afshari, B. Baradaran, R. Hajiaghaee, S. Rezazadeh, and H.Monsef-Esfahani. 2009. Suppression of nitric oxide production in activated murine peritoneal macrophages in vitro and ex vivo by Scrophularia striata ethanolic extract. J. Ethnopharmacol. 124(1):166–169. 7. Azarneoshan, F., S. Khatam Saz, and H. Sadeghi. 2009. The effects of hydro alcoholic extract of Dorema Aucheri on blood concentration of gonadotropin and androgen hormones in adult male rats. Armaghan Danesh. 14(3): 63–70. 8. Cimino, C. V., C. S. Liggieri, N. S. Priolo, M. A. Bruno, and E.Vairo-Cavalli. 2010. Arctium minus (Hill) Bernh. (Asteraceae) aspartylendopeptidases with potential application in the formulation of nutraceutical products. Mol. Med. Chem. 21:11–16. 9. Dokhani, S., T. Cottrell, J. Khajeddin, and G. Mazza. 2005 Analysis of aroma and phenolic components of selected Achillea species. Plant Foods Hum. Nutr. 60(2):55–62. 10. Ghorbani, A., F. Naghibi, and M. Mosaddegh . 2006. Ethnobotany, ethnopharmacology and drug discovery. Iranian. J. Pharm. Res. 2(2):109–118 11. Hajiaghaee, R., H. R. Monsef-Esfahani, M. R. Khorramizadeh, F. Saadat, A. R. Shahverdi, and F. Attar. 2007. Inhibitory effect of aerial parts of Scrophularia striata on matrix metalloproteinases expression. Phytother. Res. 21(12):1127–1129. 12. Jibao, C., L. Ping, Z. Xiaolan, and S. Qingde. 2006. Comparative analysis of clary sage (S. sclarea L.) oil volatiles by GC–FTIR and GC–MS. Food Chem. 99:401–407. ´ 13. Ku´zma, L., E. Bruchajzer, and H. Wysokinska. 2008. Diterpenoid production in hairy root culture of Salvia sclarea L. Z Naturforsch C. 63(7–8):621–624.


220

M. Mosaddegh et al.

14. Ku´zma, L., D. Kalemba, M. Rózalski, B. Rózalska, M. Wieckowska-Szakiel, U. ´ Krajewska, and H. Wysokinska. 2009. Chemical composition and biological activities of essential oil from Salvia sclarea plants regenerated in vitro. Molecules. 14(4):1438–1447. ˙ ˙ 15. Kuz´ma, Ł., M. Ro´zalski, E. Walencka, B. Ro´zalska, and H. Wysokin´ska. 2007. Antimicrobial activity of diterpenoids from hairy roots of Salvia sclarea:Salvipisone as a potential anti-biofilm agent active against antibiotic resistant Staphylococci. 2007. Phytomed.14:31–35. 16. Li, J., X. Liu, F. Dong, J. Xu, Y. Zheng, and W. Shan. 2010. Determination of the volatile composition in essential oil of Descurainia sophia (L.) Webb ex Prantl (Flixweed) by gas chromatography/mass spectrometry (GC/MS). Molecules 15(1):233–240. 17. Mahdavi, M., and R. Yazdanparast. 2007.Gnidilatimonoein from Daphne mucronata induces differentiation and apoptosis in leukemia cell lines. Arch. Pharm. Res. 30(2):177–181. 18. Masoudi, S., M. Jamzad, L. Attari, and A. Rustaiyan. 2003. Volatile constituents of Stachys pilifera Benth. and Stachys acerosa Boiss. from Iran. J. Essential Oil Res. 15(3):189–190 19. Mokhtari, M., M. Shariati, and H. Niknam .2008. Analgesic and anti-inflammatory effect of hydroalcoholic axtract of Dorema aucheri in male albino mice. J. Rafsanjan Univ. Med. Sci. 79(3):165–172. 20. Monsef-Esfahani, H. R., R. Hajiaghaee, A. R. Shahverdi, M. R. Khorramizadeh, and M. Amini. 2010. Flavonoids, cinnamic acid and phenyl propanoid from aerial parts of Scrophularia striata. Pharm. Biol. 48(3):333–33-6. 21. Mosaddegh, M., S .N. Ostad, F. Naghibi, and M. Hamzeloo Moghadam. 2006. The cytotoxic effects of five species of Inula against some tumor cell lines. Iranian J. Pharm. Sci. 2(4):203–208. 22. Mosaddegh, M., M. Hamzeloo Moghadam, S. Ghafari, F. Naghibi, S. N. Ostad, and R. W. Read. 2010. Sesquiterpene lactones from Inula oculus-christi. Nat. Prod. Commun. 5(4):511–514 23. Orhan, D. D., M. Aslan, G. Aktay, E. Ergun, E. Yesilada, and F. Ergun. 2003. Evaluation of hepatoprotective effect of Gentiana olivieri herbs on subacute administration and isolation of active principle. Life Sci. 72(20):2273–2283. 24. Peana, A., and M. Satta 1993. A priliminary research on the essential oils of Salvia sclarea and Salvia desoleana. Pharmacol. Res. 27:25–26. 25. Powell, E. E., G. A. Hill, B. H. Juurlink, and J. Carrier. 2005. Glucoraphanin extraction from Cardaria draba: Part 1. Optimization of batch extraction. J. Chem. Technol. Biotechnol. 80:985–991 26. Rahimifard, N., H. Hajimehdipoor, M. H. Hedayati, O. Bagheri, and A. Pishehvar, 2010. Cytotoxic effects of essential oils and extracts of some Mentha species on Vero, Hela and Hep2 cell lines. J. Med. Plants. 9:88–92. 27. Rasool, M. A., R. Khan, A. Malik, N. Bibi, and S. U. Kazmi. 2010. Structural determination of daphnecin, a new coumarinolignan from Daphne mucronata. J. Asian Nat. Prod. Res. 12(4):324–327. 28. Seol, G. H., H. S. Shim., P. J. Kim., H. K. Moon, K. H. Lee, I. Shim, S. H. Suh, and S. S. Min. 2010. Antidepressant-like effect of Salvia sclarea is explained by modulation of dopamine activities in rats. J. Ethnopharmacol. 130(1):187–190.



221

29. Sezik, E., M. Aslan, E. Yesilada, and S. Ito 2005. Hypoglycaemic activity of Gentiana olivieri and isolation of the active constituent through bioassaydirected fractionation techniques. Life Sci. 76(11):1223–1238. 30. Sharififar, F., S. Pournourmohammadi, and M. Arabnejad. 2009. Immunomodulatory activity of aqueous extract of Achillea wilhelmsii C. Koch in mice. Indian J. Exp. Biol. 47(8):668–671. 31. Souri, E., G. Amin, A. Dehmobed-Sharifabadi, A. Nazifi, and H. Farsam 2004. Antioxidant activity of sixty plants from Iran. Iranian J. Pharm. Res. 3:55–59. 32. Statistical Center of Iran. http://www.amar.org.ir 33. Sun, K., X. Li, J. M. Liu, J. H. Wang, W. Li, and Y. Sha 2005. A novel sulphur glycoside from the seeds of Descurainia sophia. J. Asian Nat. Prod. Res. 7(6):853–856. 34. Sun, K., X. Li, W. Li, J. M. Liu, J. H. Wang, and Y. Sha. 2006. A new nor-lignan from the seeds of Descurainia sophia. Nat. Prod. Res. 20(6):519–522. 35. Talib, W. H., and A. M. Mahasneh. 2010. Antiproliferative activity of plant extracts used against cancer in traditional medicine. Sci. Pharm. 78:33–45. 36. Wang, A. Q., X. K. Wang, J. L. Li, and X. Y. Cui. 2004 .Isolation and structure identification of chemical constituents from the seeds of Descurainia sophia (L.) Webb ex Prantl. Yao Xue Xue Bao. 39(1):46–51. 37. Ulubelen, A., G. Topcu, C. Eris, S. O. Nmez, M. Kartal, S. Kurucu, and C. BozokJohansson. 1994. Terpenoids from Salvia sclarea. Phytochemstry 36(4):971–974. 38. Walencka, E., S. Rozalska, H. Wysokinska, M. Rozalski, L. Kuzma, and B. Rozalska. 2007. Salvipisone and aethiopinone from Salvia sclarea hairy roots modulate staphylococcal antibiotic resistance and express anti-biofilm activity. Planta Med. 73(6):545–551. 39. Yazdanparast, R., and H. Sadeghi. 2004. Nucleic acid synthesis in cancerous cells under the effect of gnidilatimonoein from Daphne mucronata. Life Sci. 74(15):1869–1876.