Comparative evaluation of traditional prescriptions from Cichorium intybus L. for wound healing: Stepwise isolation of an active component by in vivo bioassay and its mode of activity

Ethnobotanical field surveys revealed that various parts of Cichorium intybus L. has been used for wound healing in Turkish folk medicine. The present study aimed at verifying the efficiency of various traditional prescriptions prepared from the aerial parts and roots of C. intybus experimentally and to define the components responsible from the activity by bioassay-guided procedures. Initially, wound healing activity of the aerial parts, leaves, and roots as well as ashes of either leaves or roots were investigated. Subsequently, roots of the plant were submitted to further detailed investigations. The wound healing activity of the methanolic extract, its subextracts, and fractions were evaluated by using in vivo linear incision and circular excision wound models in rats. The hydroxyproline content of the tissues treated with test ointments were also assessed for the activity evaluation. Moreover, in order to find out a possible involvement of antioxidant activity in wound healing, the test samples were also investigated by DPPH radical scavenging activity and total phenolic concentration were also determined. Additionally anti-inflammatory activity was assessed by using the method of Whittle, which is based on the inhibition of acetic acid-induced increase in capillary permeability. Through the bioassay guided fractionation one compound was isolated and its structure was elucidated by spectroscopic methods. For the determination of the activity mechanisms, the fractions were screened for hyaluronidase, collagenase and elastase enzyme inhibitory activities. Methanolic extract of C. intybus roots was found to possess potent wound healing activity. Then this extract was subjected to successive solvent extraction with n-hexane, dichloromethane (DCM), ethyl acetate and n-butanol. Each solvent extracts were also applied on the same wound models. DCM subextract was found to be the most active one and through chromatographic techniques DCM subextract was fractionated into several fractions and compound 1 was isolated as the compound being responsible from the wound healing activity. The experimental study revealed that C. intybus methanolic extract displays wound healing effect and β-sitosterol was determined as the active compound responsible from the activity. [Display omitted]