ORIGINAL PAPER Anethole and eugenol reduce in vitro and in vivo leukocyte migration induced by fMLP, LTB 4 , and carrageenan Camila Fernanda Esteva ˜o-Silva • Raquel Kummer • Fernanda Carolina Fachini-Queiroz • Renata Grespan • Gessilda Alca ˆntara Nogueira de Melo • Silmara Baroni • Roberto Kenji Nakamura Cuman • Ciomar Aparecida Bersani-Amado Received: 22 May 2013 / Accepted: 3 April 2014 / Published online: 1 May 2014 Ó The Japanese Society of Pharmacognosy and Springer Japan 2014 Abstract The aim of this study was to investigate the effect of anethole (AN) and eugenol (EUG) on leukocyte migration using in vitro chemotaxis and in situ micro- circulation assays. BALB/c mice were used for the in vitro chemotaxis assay, and Wistar rats for the in situ microcirculation assay. We evaluated (a) the in vitro leukocyte migration in response to chemotactic factors (formyl-methionyl-leucyl-phenylalanine [fMLP] and leu- kotriene B 4 [LTB 4 ]) and (b) the rolling, adhesion, and migration of leukocytes induced by an injection of car- rageenan (100 lg/cavity) into the scrotum of the animal. In the in vitro chemotaxis assay, AN and EUG at doses of 1, 3, 9, and 27 lg/ml significantly inhibited leukocyte migration when stimulated by the chemotactic agents fMLP and LTB4. In the in situ microcirculation assay, AN at doses of 125 and 250 mg/kg and EUG at a dose of 250 mg/kg significantly decreased the number of leuko- cytes that rolled, adhered, and migrated to perivascular tissue. The results indicate that AN and EUG exert inhibitory effects on leukocyte migration, highlighting their possible use to diminish excessive leukocyte migration in the inflammatory process. Keywords Anethole Á Eugenol Á Leukocyte recruitment Á Microcirculation Introduction Medicinal plants have been the basis of traditional medi- cine worldwide for hundreds of years [1], and frequently represent a major source of substances with biological activities that can be used to cure diseases [2]. The essential oils of plants and their constituents have many applications [3] and can be used in both the pharmaceutical and food industries [4, 5]. Some essential oils consist of numerous terpenoid compounds (e.g., monoterpene, ses- quiterpene, diterpene, triterpene, and tetraterpene) or phe- nylpropanoids. Phenylpropanoids are compounds formed from shikimic acid that generate p-coumaric acid and cinnamic acid. Cinnamic acid, after undergoing reduction of the side chain, generates propenylbenzene (anethole [AN]) and allylbenzene (eugenol [EUG]). An examination of the chemical structures of AN and EUG has shown that these compounds have the same biogenic origin [6], and they are considered to be structural analogues [4]. Illicium verum Hook f. is popularly known as star anise. It belongs to the Illiaceae family, which was originally distributed in tropical and subtropical areas of Asia. Trans- anethole (AN) is the major component of I. verum, corre- sponding to 90 % of the total essential oil [7]. Trans-ane- thole (1-methoxy-4-[1-propenyl]benzene) is often used in the food industry as a flavoring in beverages and food [4]. Fruits are commonly used as spices and in the treatment of gastrointestinal tract diseases [8, 9]. Studies have shown that AN has antioxidant, antibacterial [10], fungicidal [11, 12], anti-inflammatory [7, 13], anesthetic [14], and antin- ociceptive [15] effects. Eugenol is obtained from Syzygium aromaticum L. Merr. and L. M. Perry [5], or Eugenia caryophyllata L. Merr. and Perry [16], popularly known as clove. It is a large tree of the Myrtaceae family, and its vegetative cycle C. F. Esteva ˜o-Silva Á R. Kummer Á F. C. Fachini-Queiroz Á R. Grespan Á G. A. Nogueira de Melo Á S. Baroni Á R. K. N. Cuman Á C. A. Bersani-Amado (&) Laboratory of Inflammation, Department of Pharmacology and Therapeutic, State University of Maringa ´, Maringa ´, PR, Brazil e-mail: cabamado@uem.br 123 J Nat Med (2014) 68:567–575 DOI 10.1007/s11418-014-0839-7