In Vitro Antioxidant and Enzyme Inhibition Activity of Tanacetum argyrophyllum (K. Koch) Tzvelev var. argyrophyllum Extract.

Objectives: Tanacetum L. belongs to Asteraceae family and is represented by 46 species in Türkiye. Tanacetum genus is known for its insecticide and insect repellent effect. T. argyrophyllum contains sesquiterpene lactone derivatives. These compounds are responsible for its various activities, especially cytotoxic, antitumor, phytotoxic, antimicrobial, antiviral, and antifungal activity. There are not enough biological activity studies on the plant that are likely to have a wide variety of activities in terms of the compounds it contains. The aim of the present study is to evaluate various biological activities of 80% aqueous methanol extract prepared from aerial parts of T. argyrophyllum (K. Koch) Tzvelev var. argyrophyllum collected from Sivas province of Turkey. Materials and Methods: Antioxidant activity of the methanol extract was determined by 2,2'-diphenyl-1-picryl-hydrazyl and 2,2-azinobis (3-ethyl benzothiazoline-6-sulfonic acid) radical scavenging activity, total phenolic, and total flavonoid content tests. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities were investigated via Ellman's spectrometric method. Results: Total phenolic content was found as 71.67 mg/gallic acid equivalent g and total flavonoid content was 25.225 mg/quercetin equivalent g on a dry extract weight basis. In this work, AChE, BChE, and α-glycosidase enzymes were inhibited by the extract of T. argyrophyllum var. argyrophyllum. IC50 values for these enzymes were found as 266.79 µg/mL for AChE and 176.91 µg/mL for BChE. Also, the α-glycosidase activity exhibited a dose-dependent manner with increasing concentration. Conclusion: According to the results, T. argyrophyllum var. argyrophyllum can be used as an ingredient of functional foods as well as herbal products for diabetic and Alzheimer's disease patients.

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.

Bioassay-guided isolation of wound healing active compounds from Echium species growing in Turkey.

ETHNOPHARMACOLOGICAL RELEVANCE: The roots and root barks of Echium sp. have been used to treat ulcers, burns and wounds in traditional Turkish medicine. AIM OF THE STUDY: On the basis of them traditional use and literature references, four Echium species were selected for evaluation of them wound healing potential. Isolation of active component(s) from the active extracts through the bioassay guided fractionation procedures. MATERIAL AND METHODS: In vivo the wound healing activity of the plants was evaluated by linear incision experimental models. The chloroform extract of Echium italicum L. was fractionated by successive chromatographic techniques. Wound healing activity of each fraction was investigated following the bioassay-guided fractionation procedures. Moreover, the tissue samples of isolated compounds were examined histopathologically. The healing potential was comparatively assessed with a reference ointment Madecassol®, which contains 1% extract of Centella asiatica. RESULTS: Significant wound healing activity was observed from the ointment prepared with ethanol extract at 1% concentration. The ethanol root extract treated in groups of animals showed a significant increase (37.38%, 40.97% and 35.29% separately for E. italicum L, Echium vulgare L. and Echium angustifolium Miller) wound tensile strength in the incision wound model. Subfractions showed significant but reduced wound healing activity on in vivo wound models. Shikonin derivatives "Acetylshikonin", "Deoxyshikonin" and "2-methyl-n-butyrylshikonin+Isovalerylshikonin", were isolated and determined as active components of active final subfraction from E. italicum L. roots. The results of histopathological examination supported the outcome of linear incision wound models. CONCLUSION: The experimental study revealed that Echium species display remarkable wound healing activity.

In vitro cholinesterase inhibitory and antioxidant effect of selected coniferous tree species.

OBJECTIVE: To explore cholinesterase inhibitory and antioxidant effect of six coniferous trees (Abies bornmulleriana, Picea pungens, Juniperus communis, Cedrus libani, Taxus baccata, and Cupressus sempervirens var. horizantalis). METHODS: Acetone (Ace), ethyl acetate (EtOAc), and ethanol (EtOH) extracts prepared from the needles and shoots of the six coniferous trees were screened for their acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity at 100 µg/mL. Antioxidant activity of the extracts was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and N,N-dimethyl-p-phenylendiamine (DMPD) radical scavenging, metal-chelation capacity, ferric-(FRAP) and phosphomolibdenum-reducing antioxidant power (PRAP) assays. All of the assays were performed in ELISA microplate reader. Total phenol and flavonoid amounts in the extracts were determined spectrophotometrically. RESULTS: Among thirty-six extracts in total, the shoot-Ace extract of Cupressus sempervirens var. horizantalis exerted the highest inhibition against AChE [(54.84±2.51)%], while the needle-Ace extract of Cedrus libani was the most effective in inhibiting BChE [(67.54±0.30)%]. The highest DPPH radical scavenging effect, FRAP and PRAP was observed in the shoot-Ace and EtOAc extracts from Taxus baccata, whereas all the extracts showed a variable degree of scavenging effect against DPMD radical. The shoot-EtOAc extract of Cedrus libani had the highest metal-chelation capacity [(58.04±0.70)%]. The shoot extracts of Taxus baccata were determined to have the richest total phenol content, which may contribute to its marked antioxidant activity. CONCLUSIONS: The conifer species screened in this study may contain cholinesterase-inhibiting and antioxidant properties, which might be useful against Alzheimer's disease.

In vitro cholinesterase inhibitory and antioxidant effect of selected coniferous tree species

Objective: To explore cholinesterase inhibitory and antioxidant effect of six coniferous trees (Abies bornmulleriana, Picea pungens, Juniperus communis, Cedrus libani, Taxus baccata, and Cupressus sempervirens var. horizantalis). Methods: Acetone (Ace), ethyl acetate (EtOAc), and ethanol (EtOH) extracts prepared from the needles and shoots of the six coniferous trees were screened for their acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity at 100 μg/mL. Antioxidant activity of the extracts was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and N,N-dimethyl-p-phenylendiamine (DMPD) radical scavenging, metal-chelation capacity, ferric-(FRAP) and phosphomolibdenum-reducing antioxidant power (PRAP) assays. All of the assays were performed in ELISA microplate reader. Total phenol and flavonoid amounts in the extracts were determined spectrophotometrically. Results: Among thirty-six extracts in total, the shoot-Ace extract of Cupressus sempervirens var. horizantalis exerted the highest inhibition against AChE [(54.84±2.51)%], while the needle-Ace extract of Cedrus libani was the most effective in inhibiting BChE [(67.54±0.30)%]. The highest DPPH radical scavenging effect, FRAP and PRAP was observed in the shoot-Ace and EtOAc extracts from Taxus baccata, whereas all the extracts showed a variable degree of scavenging effect against DPMD radical. The shoot-EtOAc extract of Cedrus libani had the highest metal-chelation capacity [(58.04±0.70)%]. The shoot extracts of Taxus baccata were determined to have the richest total phenol content, which may contribute to its marked antioxidant activity. Conclusions: The conifer species screened in this study may contain cholinesterase-inhibiting and antioxidant properties, which might be useful against Alzheimer’s disease.

Appraisal on the wound healing and anti-inflammatory activities of the essential oils obtained from the cones and needles of Pinus species by in vivo and in vitro experimental models.

ETHNOPHARMACOLOGICAL RELEVANCE: According to ethnobotanical data, Pinus species have been used against rheumatic pain and for wound healing in Turkish folk medicine. Essential oils from the cones and needles of five different Pinus species (Pinus brutia Ten., Pinus halepensis Mill., Pinus nigra Arn., Pinus pinea L. and Pinus sylvestris L.) were evaluated for their in vivo wound healing and anti-inflammatory activities. MATERIALS AND METHODS: In vivo wound healing activity of the ointments prepared from essential oils was evaluated by linear incision and circular excision experimental wound models subsequently histopathological analysis and hydroxyproline content. Furthermore, the essential oils were screened for anti-hyaluronidase activity. 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. RESULTS: The essential oils obtained from the cones of Pinus pinea and Pinus halepensis demonstrated the highest effects on the wound healing activity models. On the other hand, the rest of the essential oils did not show any significant wound healing and anti-inflammatory activities. CONCLUSION: The experimental study revealed that essential oils obtained from the cones of Pinus pinea and Pinus halepensis display remarkable wound healing activity.

Estimation of cholinesterase inhibitory and antioxidant effects of the leaf extracts of Anatolian Ficus carica var. domestica and their total phenol and flavonoid contents.

Ficus carica var. domestica Tsch. & Rav. (common fig) is widely grown in Turkey and exported for its edible fruits. In this study, the n-hexane, chloroform, acetone, methanol, n-butanol, and water extracts of the leaves of F. carica var. domestica were screened for their cholinesterase inhibitory and antioxidant activities. Cholinesterase inhibition against acetyl- (AChE) and butyrylcholinesterase (BChE) was measured by the spectrophotometric method of Ellman at concentrations of 25, 50, and 100 microg/mL., while antioxidant activity was tested using three in vitro methods; 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, metal-chelation capacity, and ferric-reducing antioxidant power (FRAP). Total phenol and flavonoid contents of the extracts were determined spectrophotometrically. Our results revealed that the n-hexane and acetone extracts exerted a notable inhibition against both AChE (62.9 +/- 0.9% and 50.8 +/- 2.1%, respectively) and BChE (76.9 +/- 2.2% and 45.6 +/- 1.3%, respectively). However, they had low activity in the antioxidant tests. The chloroform extract was found to be the richest in total flavonoid content (252.5 +/- 1.1 mg/g quercetin equivalent), while the n-butanol extract had the highest total phenol amount (85.9 +/- 3.2 mg/g extract gallic acid equivalent).

Flavonoids with anti-Helicobacter pylori activity from Cistus laurifolius leaves.

Cistus laurifolius flower buds are used traditionally in folk medicine against gastric ailments. In a prior study we showed that the chloroform extract of Cistus laurifolius had a potent anti-ulcer activity. It has been known that there is a causal relationship between peptic ulcer and Helicobacter pylori infection. Then in a previous study, we demonstrated that chloroform extract of Cistus laurifolius possessed a significant anti-Helicobacter pylori activity. We designed this study to isolate and define the active component(s) involved in the anti-Helicobacter pylori activity of the extract through activity-guided fractionation procedures. The chloroform extract was fractionated by using various chromatography techniques, i.e., Sephadex LH-20 column chromatography and preparative thin layer chromatography and six compounds were isolated (1-6). Each of these six compounds' anti-Helicobacter pylori activity was tested in vitro and was measured as minimum inhibition concentration (MIC) values by using agar dilution method. The compound 2 had the highest activity against Helicobacter pylori (MIC 3.9 microg/mL). Its chemical structure was elucidated as quercetin 3-methyl ether (isorhamnetin) by various spectroscopic techniques. We believe that the therapeutic effect of Cistus laurifolius in ulcer is at least partially related to its effect on Helicobacter pylori. We hope that the isolated flavonoid having anti-Helicobacter pylori activity ultimately can be utilized as an alternative or additive agent to the current therapy.

Anti-ulcerogenic activity of some plants used as folk remedy in Turkey.

Five herbal remedies used as gastroprotective crude drugs in Turkey were assessed for anti-ulcerogenic activity using the EtOH-induced ulcerogenesis model in rat. The crude drugs investigated comprises fruits of Elaeagnus angustifolia L. (Elaeagnaceae), fresh fruits of Hibiscus esculentus L. (Malvaceae), fresh roots of Papaver rhoeas L. (Papaveraceae), leaves of Phlomis grandiflora H.S. Thomson (Lamiaceae) and fresh fruits of Rosa canina L. (Rosaceae). Extracts were prepared according to the traditional indications of use. Under our experimental conditions, all extracts exhibited statistically significant gastroprotective effect with better results for Phlomis grandiflora and Rosa canina (100%). At the concentration under study, both crude drugs were more effective than the reference compound misoprostol at 0.4 mg/kg. Even the weakest anti-ulcerogenic effect observed for Papaver rhoeas roots was found statistically potent (95.6%). Histopathological studies confirmed the results of the in vivo test.

In vivo gastroprotective effects of five Turkish folk remedies against ethanol-induced lesions.

Through evaluation of the data accumulated in Data Bank of Turkish Folk Remedies (TUHIB), five plant remedies, which are used to treat stomach ache were selected to test for their anti-ulcerogenic potency. In order to confirm the claimed activities, either decoction or methanol extracts were prepared from the roots of Asphodelus aestivus and Cichorium intybus, herbs of Equisetum palustre and Viscum album ssp. album and fruits of Laurus nobilis, according to their folkloric application way and tested for their effects on ethanol-induced gastric ulcer model in rats. Pharmacological experiments clearly demonstrated that the relevant extracts of all the plants given orally showed significant stomach protection against this model of ulcerogenesis. Results were further evaluated by using histopathological techniques.