A Comparison Study of the Effects of Echinacea purpurea Ethanolic Extract and Mesna on Cyclophosphamide-Induced Macroscopic Fetal Defects in Rats

Document Type: Original Article


1 Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Shahid Chamran University, Ahwaz, Iran

2 Basic Sciences Group, Faculty of Veterinary Medicine, Shahid Chamran University, Ahwaz, Iran


There are some reports that the teratogenic effects of cyclophosphamide (CPA) can be prevented by application of antioxidant drugs and stimulation of the maternal immune system. Echinacea purpurea extract is antioxidative and immunomodulator drug. Mesna (Sodium 2-mercaptoethane sulfonate) is used for decreasing side effects of CPA, especially hemorrhagic cystitis. In this study, we compared the prophylactic effects of mesna and Echinacea extract on teratogenic effects of CPA.
Materials and Methods
This study was performed on 32 pregnant rats that were divided into 4 groups. The first group (control group) received normal saline and the other groups received CPA (15 mg/kg intraperitoneally) on 13th day of gestation. Mesna and E. purpurea extracts were administrated at doses of 100 and 400 mg/kg by IP injection, respectively, along with it and 12 hr later, after CPA injection. Rats were dissected on day 20 of gestation, embryos harvested and after determination of gross malformations they were stained by Alizarin red-Alcian blue method.
Cleft palate incidence was 38.46, 30.77 and 14.28% in fetuses of rats that received only CPA, CPA with mesna and CPA with Echinacea extract, respectively. In addition, skeletal anomalies incidence including limbs, vertebra, sternum, and scapula defects were decreased by Echinacea extract.
E. purpurea has significant effect on preventing CPA-induced malformations and better prophylactic effect than mesna on cases like CPA-induced cleft palate.


1.Giavini E, Menegola E. Gene-teratogen chemically induced interactions in congenital malformations. Biol Neonate 2004; 85:73-81.

2.Finnell RH. Teratology: general considerations and principles. J Allerg Clin Immunol 1999; 103:337-342.

3.Moore KL, Persaud TVN. The developing human clinically oriented embryology. 8th ed. Saunders Elsevier; 2008. 

4.De Santis M, Straface G, Carducci B, Cavaliere AF, De santis L, Lu echese A, et al. 2004. Risk of drug- induced congenital defects. Eur J Obestet Gynecol Reprod Biol 2004; 117:9-10.

5.Orup HI, Holmes LB, Keith DA, Coull BA. 2003. Craniofacial skeletal deviations following in utero exposure to the anticonvulsant phenytoin. Monotherapy and polytherapy orthod. Orthod Craniofac Res 2003; 6: 2-19.

6.Prater MR, Zimmerman KL, Lee Ward D, Holladay SD. Reduced birth defects caused by maternal immune stimulation in methylnitrosourea-exposed mice: Association with placental improvement. Birth Defects Res 2004; 70: 862-869.

7.Holladay SD, Sharova LV, Punareewattana K, Hrubec TC, Gogal RM, Prater MR. Maternal immune stimulation in mice decreases fetal malformations caused by teratogens. Int Immunopharmacol 2002; 2:25- 332.

8.Holladay SD, Sharova LV, Smith BJ, Gogal RM, Ward DL, Blaylock BL. Nonspecific stimulation of the maternal immune system. I. Effects on teratogen-induced fetal malformations. Teratology 2000; 62:413-419.

9.Syska E, Schmidt R, Schubert J. The time of palatal fusion in mice: a factor of strain susceptibility to teratogens. J Craniomaxillofac Surg 2004; 32: 2-4.

10.Toder V, Strassburger D, Irlin Y, Carp H, Pecht M,Trainin N. Nonspecific immunopotentiators and pregnancy loss: complete Freund adjuvant reverses high fetal resorption rate in CBA x DBA/2 mouse combination. Am J Repord Immunol 1990; 24: 63-66.

11.Carp HJ, Toder V, Torchinsky A, Portuguese S, Lipitz S, Gazit E, et al. Allogeneic leukocyte immunization after five or more miscarriage immunotherapy trialists group. Huma Reprod 1997; 12:250-255.

12.Ho HN, Chen SU, Yang YS, Huang SC, Lee TY, Gill TJ. Age, environment, and lymphocyte immunization influence the spontaneous resorption rate in the CBA/J x DBA/2J mouse model. Am J Repord Immunol 1994; 31:47-51.

13.Nomura T, Hata S, Kusafuka T. Suppression of developmental anomalies by maternal macrophages in mice. J Exp Med 1990; 172:1325-1330.

14.Ivnitsky I, Torchinsky A, Gorivodsky M, Zemliak I, Orenstein H, Savion S, et al. 1998. TNF-alpha expression in embryos exposed to a teratogen. Am J Reprod Immunol 1998; 40:431-440.

15.Savion S, Brengauz-Breitmann M, Torchinsky A, Toder V. A possible role for granulocyte macrophage-colony stimulating factor in modulating teratogen-induced effects. Teratog Carcinog Mutagen 1999; 19:171-182.

16.Hardman JG, Limbird Lee E. The pharmacological basis of therapeutics. 10th ed. McGrow-Hill; 2001.

17.Slott VL, Hales BF. Sodium 2-mercaptoethane sulfonate protection against cyclophosphamide-induced teratogenicity in rats. Toxicol Appl Pharmacol 1986; 82:80-86.

18.Barrett B. Medicinal properties of Echinacea. Phytomedicine 2003; 10: 66-86.

19.Khaksary Mahabady M, Ranjbar R, Arzi A, Papahn AA, Najafzadeh H. 2006. A comparison study of effects of Echinacea extract and levamisole on phenytoin-induced cleft palate in mice. Regul Toxicol Pharmacol 2006; 46:163-166.

20.Yolanda P. Alizarin staining. Laboratory exercises in developmental biology. Academic Press limited; 1993.

21.Ivnitsky I, Torchinsky A, Savion S, Shepshelovich J, Orenstein H, Toder V, et al. TGF beta 2 in embryos with inborn anomalies: effect of maternal immunopotentiation. Am J Reprod Immunol 2001; 45: 41-51.

22.Winn LM, Wells PG. Maternal administration of superoxide dismutase and catalase in phenytoin teratogenicity. Free Radic Biol Med 1999; 26: 266-274.

23.Sharova LV, Gogal RM, Sharov AA, Chrisman MV, Holladay SD. Stimulation in urethane- exposed pregnant mice increase expression level of spleen leukocyte genes for TGF beta 3 GM- CSF and other cytokines that may play a role reduced chemical - induced birth defects. Int Immunopharmacol 2002; 10:1477-1489.

24.Skarek T, Tynecka Z, Glowniak K, Lutostanska E. Echinacea L.-Inducer of interferons. Herba Polonica 1996; 42:110-117.

25.Bukovsky M, Vaverkova S, Kostalova D. Immunomodulating activity of Echinacea gloriosa L., Echinacea angustifolia DC, and Rudbeckia speciosa wenderoth ethanol-water extracts. Pol J Pharmacol 1995; 4:175-177.

26.Mishima S, Saito K, Maruyama H, Inoue M, Yamashita T, Ishida T, et al. Antioxidant and immuno-enhancing effects of Echinacea purpurea. Biol Pharm Bull 2004; 27:1004-1009.

27.Linde K, Barrett B, Wolkart K, Bauer R, Melchart D. Echinacea for preventing and treating the common cold. Cochrane Database Syst Rev 2006; 25:CD000530.

28.Matthias A, Banbury L, Stevenson LM, Bone KM, Leach DN, Lehmann RP. Alkylamides from Echinacea modulate induced immune responses in macrophages. Immunol Invest 2007; 36:117-130.

29.Matthias A, Banbury L, Bone KM, Leach DN, Lehmann RP. 2008. Echinacea alkylamides modulate induced immune responses in T-cells. Fitoterapia 2008; 79: 53-58.