Bioactivities of Ethanolic Extract and its Fractions of Cistus laurifolius L. (Cistaceae) and Salvia wiedemannii Boiss. (Lamiaceae) Species.

BACKGROUND: Cistus laurifolius L. (Cistaceae) and Salvia wiedemannii Boiss. (Lamiaceae) have been used for treatment of some illnesses in Turkish folk medicine. In the present study, the ethanolic extract and its fractions obtained using re-extraction by hexane (Hx), chloroform (CHCl3), butanol, and remaining-water (r-H2O) of C. laurifolius were screened for their in vitro bioactivities. MATERIALS AND METHODS: Activities were determined against both standard and the isolated strains of Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Enterococcus faecalis, as well as yeasts such as Candida albicans and Candida parapsilosis by microdilution method. Also, antiviral activity of C. laurifolius and S. wiedemannii extracts were tested on herpes simplex virus-1 (HSV-1) and parainfluenza-3 (PI-3) using Madin-Darby bovine kidney and vero cell lines. RESULTS: Tested extracts of C. laurifolius (minimum inhibitory concentration 32 µg/mL) exerted a strong antimicrobial activity against Gram-negative bacteria of E. coli, P. mirabilis, K. pneumoniae, and A. baumannii. CONCLUSION: The Hx extract of C. laurifolius (cytopathogenic effect of 32-8 µg/mL) had antiviral activity on PI-3. Also, the r-H2O, CHCl3, and ethanol extracts (16-<0.25 µg/mL) of S. wiedemannii had significant antiviral activity on HSV-1, same as control. SUMMARY: The objective of this study was to evaluate the bioactivity of plant extracts used in folk medicineEthanolic extract and its fractions obtained using re-extraction by hexane (Hx), chloroform (CHCl3), butanol, and remaining-water (r-H2O) of Cistus laurifolius L. (Cistaceae) and Salvia wiedemannii Boiss. (Lamiaceae) were screened against both standard and the isolated strains of E. coli, P. aeruginosa, P. mirabilis, K. pneumoniae, A. baumannii, S. aureus, E. faecalis, C. albicans and C. parapsilosis by microdilution methodAntiviral activity were tested on HSV-1 and PI-3 using MDBK and Vero cell linesExtracts of C. laurifolius exerted a strong antimicrobial activity against E. coli, P. mirabilis, K. pneumoniae, and A. baumannii(MIC; 32 µg/mL)The Hx extract of C. laurifolius had antiviral activity on PI-3 (CPE; 32-8 µg/mL). Also, the r-H2O, CHCl3, and ethanol extracts (16-<0.25 µg/mL) of S. wiedemannii had significant antiviral activity.

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.

ETHNOPHARMACOLOGICAL RELEVANCE: Ethnobotanical field surveys revealed that various parts of Cichorium intybus L. has been used for wound healing in Turkish folk medicine. AIM OF THE STUDY: 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. MATERIAL AND METHODS: 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. RESULTS: 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. CONCLUSION: The experimental study revealed that C. intybus methanolic extract displays wound healing effect and ß-sitosterol was determined as the active compound responsible from the activity.

Efficacy of Daphne oleoides subsp. kurdica used for wound healing: identification of active compounds through bioassay guided isolation technique.

ETHNOPHARMACOLOGICAL RELEVANCE: In Turkish traditional medicine, the aerial parts of Daphne oleoides Schreber subsp. kurdica (DOK) have been used to treat malaria, rheumatism and for wound healing. The aim was to evaluate the ethnopharmacological usage of the plant using in vivo and in vitro pharmacological experimental models, and to perform bioassay-guided fractionation of the 85% methanolic extract of DOK for the isolation and identification of active wound-healing component(s) and to elucidate possible mechanism of the wound-healing activity. MATERIALS AND METHODS: In vivo wound-healing activity was evaluated by the linear incision and the circular excision wound models. Anti-inflammatory and antioxidant activities, which are known to support the wound healing process, were also assessed by the Whittle method and the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging assays, respectively. The total phenolic content of the extract and subextracts was estimated to establish any correlation between the phenolic content and the antioxidant activity. The methanolic extract of DOK was subjected to various chromatographic separation techniques leading to the isolation and identification of the active component(s). Furthermore, in vitro hyaluronidase, collagenase and elastase enzymes inhibitory activity assays were conducted on the active components to explore the activity pathways of the remedy. RESULTS: After confirmation of the wound-healing activity, the methanolic extract was subjected to successive solvent partitioning using solvents of increasing polarity creating five subextracts. Each subextract was tested on the same biological activity model and the ethyl acetate (EtOAc) subextract had the highest activity. The EtOAc subextract was subjected to further chromatographic separation for the isolation of components 1, 2 and 3. The structures of these compounds were elucidated as daphnetin (1), demethyldaphnoretin 7-O-glucoside (2) and luteolin-7-O-glucoside (3). Further in vivo testing revealed that luteolin-7-O-glucoside was responsible for the wound-healing activity of the aerial parts. It was also found to exert significant anti-inflammatory, antioxidant, anti-hyaluronidase and anti-collagenase activities. CONCLUSION: The present study explored the wound-healing potential of Daphne oleoides subsp. kurdica. Through bioassay-guided fractionation and isolation techniques, luteolin-7-O-glucoside was determined as the main active component of the aerial parts. This compound exerts its activity through inhibition of hyaluronidase and collagenase enzymes activity as well as interfering with the inflammatory stage.

Assessment of anti-inflammatory and antinociceptive activities of Olea europaea L.

Leaves and fruits of Olea europaea L. (Family Oleaceae) are used for the treatment of various kinds of diseases, i.e., rheumatism and hemorrhoids, and as a vasodilator in vascular disorders for ages in folk medicine. In order to evaluate this information, anti-inflammatory and antinociceptive activities of the ethanolic and n-hexane extracts prepared from the fruits of O. europaea were investigated. For the anti-inflammatory activity, the carrageenan-induced hind paw edema and acetic acid-induced increased capillary permeability models were used, whereas for the antinociceptive activity, the p-benzoquinone-induced writhing and hot plate tests in mice were used. The results revealed that the ethanolic extract did not show a significant anti-inflammatory or analgesic activity, whereas the n-hexane extract displayed 12.7-27.8% inhibition on the carrageenan-induced hind paw edema model at the 400 mg/kg dose, without inducing any apparent acute toxicity as well as gastric damage. Further studies are warranted to define and isolate the active anti-inflammatory and antinociceptive components from this plant, which may yield safe and effective agents to be used in the treatment of inflammatory disorders.

Investigations on the in vivo wound healing potential of Hypericum perforatum L.

ETHNOPHARMACOLOGICAL RELEVANCE: Olive oil extract of the flowering aerial parts of Hypericum perforatum L. (Hypericaceae) is a popular folk remedy for the treatment of wounds in Turkey. AIM OF THE STUDY: In order to prove the claimed utilization of the plant, the effects of the extracts and the fractions were investigated by using bioassay-guided procedures. For the wound healing activity assessment, in vivo excision and incision wound models were applied. For the anti-inflammatory activity, an in vivo model, based on the inhibition of acetic acid-induced increase in capillary permeability was used as well. Moreover, a parallel study was run on Hypericum scabrum L., which is a widespread species of the gender but not known as a folk remedy for wound healing, to provide a preliminary data to compare and emphasize the selection of correct plant species. RESULTS: Initial investigations proved that the olive oil extract of Hypericum perforatum has a significant wound healing effect on excision (5.1-82.6% inhibition) and circular incision (20.2-100.0% inhibition) wound models. In order to determine the active wound healing ingredient(s), aerial parts of the plant was extracted with ethanol, noteworthy wound healing activity profile was observed with the wound models; between 18.3% and 95.6% in excision model and from 13.9% to 100.0% inhibitions in incision model were determined. The ethanolic extract was then submitted to successive solvent extractions with n-hexane, chloroform and ethyl acetate (EtOAc). Each solvent extract was also applied on the same wound models, consequently, EtOAc subextract was found to be the most active one by inhibiting wounds between 17.9% and 100.0% in excision model, subsequently between 9.4% and 100.0% in incision model. However, all subfractions obtained from the EtOAc subextract using Sephadex LH-20 column chromatography showed wound healing activity not more than the whole EtOAc subextract, which revealed that a possible synergistic activity that might be questioned. Among the active Sephadex fractions, Fr. A further yielded hyperoside, isoquercitrin, rutin and (-)-epicatechin and Fr. B yielded hypericin as the major components. Moreover, a dose-dependent anti-inflammatory activity was found for the ethanol extract, EtOAc subextract and Sephadex fractions of Hypericum perforatum. These results suggest that anti-inflammatory activity of the active fractions might have a contributory role in the wound healing effect of the plant. CONCLUSION: Results of the present study have proved that aerial parts of Hypericum perforatum possess remarkable wound healing and anti-inflammatory activities supporting the folkloric assertion of the plant in Turkish folk medicine. Flavonoids [hyperoside, isoquercitrin, rutin and (-)-epicatechin] and naphthoquinones (hypericins) were found as the active components of Hypericum perforatum. On the other hand, ethanol extract of Hypericum scabrum showed neither remarkable wound healing nor anti-inflammatory activity demonstrating the importance of correct plant species selection in therapeutic applications.