An ethnopharmacological study on Verbascum species: From conventional wound healing use to scientific verification

doi:10.1016/j.jep.2010.08.004

Journal of Ethnopharmacology

Volume 132, Issue 2, 11 November 2010, Pages 408-413

https://doi.org/10.1016/j.jep.2010.08.004 Get rights and content

Abstract

Aim of the study

The leaves, flowers, and whole aerial parts of Verbascum L. (Scrophulariaceae) species are used to treat eczema and other types of inflammatory skin conditions and as a desiccant for wounds in Turkish traditional medicine. In the present study, the methanolic extracts of 13 Verbascum species growing in Turkey, including Verbascum chionophyllum Hub.-Mor., Verbascum cilicicum Boiss., Verbascum dudleyanum (Hub.-Mor.) Hub.-Mor., Verbascum lasianthum Boiss., Verbascum latisepalum Hub.-Mor., Verbascum mucronatum Lam., Verbascum olympicum Boiss., Verbascum pterocalycinum var. mutense Hub.-Mor., Verbascum pycnostachyum Boiss. & Heldr., Verbascum salviifolium Boiss., Verbascum splendidum Boiss., Verbascum stachydifolium Boiss. & Heldr and Verbascum uschackense (Murb.) Hub.-Mor. were assessed for their in vivo wound healing activity.

Material and methods

In vivo wound healing activity of the plants were evaluated by linear incision and circular excision experimental models subsequently histopathological analysis. The healing potential was comparatively assessed with a reference ointment Madecassol^®, which contains 1% extract of Centella asiatica.

Results

The methanolic extracts of Verbascum olympicum, Verbascum stachydifolium and Verbascum uschackense demonstrated the highest activities on the both wound models. Moreover, the methanolic extracts of Verbascum latisepalum, Verbascum mucronatum, and Verbascum pterocalycinum var. mutense were found generally highly effective. On the other hand, the rest of the species did not show any remarkable wound healing effect. Results of the present study support the continued and expanded utilization of these plant species employed in Turkish folk medicine.

Conclusion

The experimental study revealed that Verbascum species display remarkable wound healing activity.

Graphical abstract

The leaves, flowers, and whole aerial parts of Verbascum L. (Scrophulariaceae) species are used to treat eczema and other types of inflammatory skin conditions for desiccating wounds in traditional Turkish medicine.

Introduction

The leaves and flowers of Verbascum densiflorum Bertol., Verbascum phlomoides L. and Verbascum thapsus L. are reported to have expectorant, mucolytic and demulcent properties which are used to treat respiratory disorders such as bronchitis, dry coughs, tuberculosis and asthma in Turkish traditional medicine. The species are also used to treat haemorrhoids, rheumatic pain, superficial fungal infections, wounds and diarrhoea, and have inhibitory activities against the murine lymphocytic leukaemia and influenza viruses A2 and B. The oil prepared from the flowers is used to help soothe earache and is applied externally for eczema and other types of inflammatory skin conditions. These species are reported to be mildly diuretic and to have a soothing and anti-inflammatory effect on the urinary tract, as well as acting as a mild sedative. They are traditionally consumed as a tea to relieve abdominal pains (Baytop, 1999, Ucar Turker and Camper, 2002, Ucar Turker and Gurel, 2005).

The aerial parts of Verbascum pumilum Boiss. and Heldr. have been used for the treatment of anal fistula. The dried and powdered leaves have been used as a desiccant for wounds, and the decocted leaves have been used for abdominal pain and bronchitis, taken as a tea. The flowers of Verbascum orientale (L). All have been used for pruritic conditions in urogenital organs (Sezik et al., 2001).

The roots and the aerial parts of Verbascum cheiranthifolium Boiss. var. cheiranthifolium and the aerial parts of Verbascum chrysochaete Stapff have been used for eczema, rheumatism, earache, haemorrhoids and menstrual pain. The flowers of Verbascum lasianthum as well as the flowers and the leaves of Verbascum symes Murb. et Rech fil. are reported to be used for haemorrhoids in the southwest Anatolia (Tuzlaci and Erol, 1999, Gurhan and Ezer, 2004). In order to evaluate their folkloric utilization, in vivo anti-inflammatory and antinociceptive activities of Verbascum lasianthum Boiss. ex Bentham flowers were investigated in our previous studies. Methanolic extract of the flowers, its secondary metabolites, aucubin and ilwensisaponin A were shown to possess significant inhibitory activity in the carrageenan-induced hind paw edema model and in p-benzoquinone-induced writhings in mice, per os without inducing any apparent acute toxicity or gastric damage (Kupeli et al., 2007a, Kupeli et al., 2007b). Moreover, antinociceptive and anti-inflammatory activities of seven endemic Verbascum species including Verbascum chionophyllum Hub.-Mor., Verbascum cilicicum Boiss., Verbascum dudleyanum (Hub.-Mor.) Hub.-Mor., Verbascum latisepalum Hub.-Mor., Verbascum pycnostachyum Boiss.& Heldr., Verbascum salviifolium Boiss., Verbascum splendidum Boiss. were investigated by our research groups. The methanolic extracts of the flowers of Verbascum chionophyllum and Verbascum pycnostachyum, the aerial parts of Verbascum latisepalum and Verbascum salviifolium displayed significant antinociceptive and anti-inflammatory activity at 200 mg/kg dose, per os, without inducing any apparent acute toxicity as well as gastric damage (Tatli et al., 2008).

The present study is aimed to investigate the possible wound healing effect of the methanol extracts of 13 Verbascum species using in vivo experimental models i.e., linear incision and circular excision wound models.

Section snippets

Plant material

Plant materials were collected from different localities between 2000 and 2008 in Turkey. Voucher specimens were authenticated by Prof. Dr. Hayri Duman (Gazi University, Faculty of Science, Etiler, Ankara, Turkey) and were deposited at the Herbarium of the Pharmacognosy Department, Faculty of Pharmacy, Hacettepe University and the Herbarium of Botany Department, Faculty of Science and Art, Gazi University, Ankara, Turkey. Collection sites, parts used and herbarium numbers of the selected

Results and discussion

In this study, the methanol extracts of 13 Verbascum L. species growing in Turkey were investigated for their in vivo wound healing activity which was assessed by linear incision and circular excision wound models. More to the point, skin samples were also evaluated histopathologically. The experimental results were given in Table 2, Table 3, Table 4.

As shown in Table 2, topical application of the ointment prepared with the methanolic extracts of Verbascum stachydifolium, Verbascum uschackense

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Indeed, with years, the extensive growth of pharmaceutical, nutraceutical, cosmeceutical, or food industries have significantly increased the demand of medicinal plants and their respective bioactive compounds [1]. In Turkish traditional medicine, the leaves, flowers, and whole aerial parts of Verbascum L. species are used to manage eczema and other types of inflammatory skin disorders and as a desiccant for wounds [2]. A total of 1364 plants are recorded in this genus according to The Plant List (2013), whereby 116 are accepted, 206 are synonyms and 1055 are unassessed [3].
Members from the Verbascum genus are well appraised for their traditional uses against a variety of ailments including anti-inflammatory related disorders. This study thus attempts at validating the biological properties of two underexplored Verbascum members namely Verbascum euphraticum Benth. (V. euphraticum) and Verbascum oocarpum Murb. (V. oocarpum) in terms of their antioxidant and enzymatic inhibitory activities. Concentration of specific bioactive compounds was also established and molecular dynamic analysis were conducted on amassed data. Results showed that total phenolic and total flavonoid contents were higher for the methanolic extract of V. oocarpum compared to the methanolic extract of V. euphraticum. Phytochemical quantification revealed that the extraction solvents influenced the phytochemical composition of the extracts. Derivatives of phenolic acids such as 3-caffeoylquinic acid and 3,5-dicaffeoylquinic acid together with some phenolic acids i.e., gallic, vanillic, caffeic, syringic, p-coumaric and ferulic acid, have been detected in the extracts of both Verbascum species. Methanolic extracts of V. euphraticum and V. oocarpum yielded the highest total content of monitored compounds resulting in 162.758 and 3716.861 μg/g of dried weight extract, respectively. The methanolic extracts of both plants showed highest antioxidant potential while the ethyl acetate and hexane extracts were potent inhibitors of cholinesterase, α-amylase and α-glucosidase. Molecular dynamic results indicated that vanillic acid is able to make strong interactions with copper ions of tyrosinase, resulting into an effective anti-tyrosinase activity. In light of the above, both Verbascum species could be considered for future phytopharmaceutical development.
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For instance, they are acknowledged as a remedy for respiratory tract infections, especially in case of irritating coughs, accompanied by bronchial congestions (Kahraman et al., 2010). Furthermore, the whole aerial parts, leaves, and flowers of Verbascum species are used to cure eczema, skin inflammations, or as wound desiccants (Süntar et al., 2010). Other uses as astringent, antiseptic, emollient, demulcent, expectorant, narcotic, sedative, diuretic, antimalarial, and as a treatment for migraine, tumors including asthma have also been reported (Tatli and Akdemir, 2006).
The genus Verbascum, commonly known as "mullein", bears several species traditionally used to treat various infections, inflammations and wounds. In this study, extracts from V. bombyciferum, an endemic Verbascum species from Turkey, obtained by different solvents and extraction techniques, were investigated for their bioactive contents using spectrophotometry and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and evaluated for their antioxidant, as well as enzyme inhibitory potentials. While spectrophotometric determination revealed the homogenizer assisted extraction/methanolic extract to yield the highest total phenolic and flavonoid contents (41.80 ± 0.70 mg GAE/g and 39.44 ± 0.30 mg RE/g, respectively), the most significant total bioactive compounds (polyphenols) in the amount of 272.862 mg kg⁻¹, established by HPLC-MS/MS analysis, was noted for the maceration/methanolic extract. Overall, the extracts showed varied antioxidant potency, displaying free radical scavenging ability (DPPH assay: 1.04–51.33 mg TE/g; ABTS assay: 80.04–85.60 mg TE/g), reducing power (CUPRAC: 51.48–183.26 mg TE/g; FRAP: 36.33–90.04 mg TE/g; metal chelating activity (18.48–19.80 mg EDTAE/g) and total antioxidant capacity (phosphomolybdenum assay: 0.78–1.92 mmol TE/g). However, extract of homogenizer assisted extraction/methanol exhibited the highest antioxidant activity in most assays. Only the maceration/methanolic extract inhibited all the studied enzymes (acetylcholinetserase: 2.02 ± 0.20 mg GALAE/g; butyrylcholinesterase: 4.57 ± 0.09 mg GALAE/g; tyrosinase 21.44 ± 3.41 mg KAE/g and amylase: 0.46 ± 0.01 mmol ACAE/g). This study is the first attempt to highlight the potential industrial applications of the endemic plant V. bombyciferum, as a natural source of bioactive compounds that could be considered for future development of phytopharmaceuticals, targeting specific oxidative stress-linked diseases including diabetes, skin hyperpigmentation, and neurological disorders.
GC-MS and LC-ESI-MS analysis of biologically active fractions from Verbascum letourneuxii; efficient protocol for in vitro propagation
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Numerous species of the genus Verbascum have been reported to possess a wide range of biological activities and various medicinal properties. Some of the best reported properties are anti-inflammatory effect (Tatli and Akdemir, 2006; Speranza et al., 2009), immunostimulants (Vijayavitthal et al., 1998), cytotoxicity (Talib and Mahasneh, 2010 a,b), wound healing activity (Korkina et al., 2007; Süntar et al., 2010), antiviral activity (Zgórniak-Nowosielska et al., 1991), antimicrobial effect (Senatore et al., 2007; Amirnia et al., 2011) and antioxidant activity (Sezik et al., 2001; Turker and Camper, 2002; Öztürk et al., 2019). Plant tissue culture is an important conservation technique for plant material production.
Verbascum species (family: Scrophulariaceae) have a wide range of medicinal properties; one of these species is Verbascum letourneuxii which is an important rarely distributed plant in Egypt. The antioxidant and cytotoxic activities of the 70% aqueous methanol extract and the four successive fractions (diethyl ether, ethyl acetate, chloroform and methanol) were evaluated. The results of the DPPH assay (100 μg/mL) suggested differential scavenging properties as ordered in descending: methanol > aqueous methanol > diethyl ether > chloroform > ethyl acetate with % values of 85.7, 80, 76, 64 and 23, respectively, compared to ascorbic acid (98.4%). Moreover, the diethyl ether and the chloroform fractions showed significant cytotoxic effects (100 μg/mL). The diethyl ether fraction showed IC₅₀ 52.5 μg/mL against HepG2, while the chloroform fraction revealed IC₅₀ 70.1, 63.6 and 73.4 (μg/mL) against HepG2, MCF-7 and PC3 compared to doxorubicin with IC₅₀ 27.4, 28.2 and 28.7 (μg/mL), respectively. The analysis of the GC/MS for diethyl ether and chloroform fractions led to detection and recognition of 53 chemical compounds, while the LC–ESI–MS analysis of the MeOH fraction revealed the identification and characterization of 21 compounds (eight iridoid glycosides, five phenylethanoids, five phenylpropanoids and three flavonoids). Additionally in this study, a first protocol for in vitro propagation of V. letourneuxii was established for conservation of this plant.
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In this line, activities of these plants used in traditional medicine have been widely investigated to understand the molecular basis and to extend their clinical use. Verbascum spp., which are used in the treatment of burns, hemorrhoids, respiratory and digestive system diseases in traditional medicine, consists 360 species (Süntar et al. 2010; Georgiev et al. 2011). In Turkey flora, 232 species are found and of those, 196 species are restricted to Turkey (Kozan et al. 2011).
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While fresh Clematis vitalba aerial parts have long been considered toxic and the plant is well known to be a revulsive and a skin irritant because of its protoanemonine content (Roth et al., 1994), its extracts have shown anti-inflammatory, anti-nociceptive, antipyretic, antimicrobial, and anti-obesity effects (Buzzini and Pieroni, 2003; Yesilada and Küpeli, 2007; Marrelli et al., 2013). The external use of Verbascum spp. for treating wounds has also been observed in Turkish folk medicine and its potent activity well confirmed in in-vivo studies, but only for a few species of the genus (Süntar et al., 2010). Fraxinus excelsior bark tea as a diuretic; this possible, but not yet sufficiently proven, activity has been addressed by very few studies (Eddouks et al., 2005; Wright et al., 2007, and references therein).
Ethnobiological research in SE Europe is especially important for providing concrete insights aimed at developing small-scale markets of local medicinal plants and food products.
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Inhibition of the DHT-induced PSA secretion by Verbascum xanthophoeniceum and Serenoa repens extracts in human LNCaP prostate epithelial cells
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is an endemic mullein plant species (genus Verbascum, Scrophulariaceae) of the Balkans and some parts of Turkey, particularly enriched in iridoids- and phenylethanoids. In Bulgarian and worldwide folk-medicine, Verbascum species have a traditional use in the treatment of inflammatory disorders of skin and epithelial tissues of the respiratory, gastro-intestinal and uro-genital tracts (Ivanov et al., 1971; Turker and Gurel, 2005; EMEA, 2009; Suntar et al., 2010). Verbascum species contain several bioactive compounds, including flavonoids and phenylethanoid glycosides among others, whose anti-inflammatory, anti-oxidative, anti-tumor and immune-stimulatory activities (Talib and Mahasneh, 2010; Georgiev et al., 2011a, 2011b) have been reported so far.
Verbascum xanthophoeniceum is a mullein plant, typical of Balkan region and some parts of Turkey, traditionally used as phytotherapeutic agent due to its anti-inflammatory properties. It is rich in phenylethanoid and iridoid metabolites whose anti-inflammatory properties are under characterization.
The role of Verbascum xanthophoeniceum crude methanolic extract and its isolated phenylethanoid glycoside verbascoside have been evaluated, in comparison to a saw palmetto extract, on a human in vitro model of androgen-regulated prostate epithelium, the LNCaP cell line. Cytotoxicity and DHT-induced free and total PSA secretion have been thoroughly studied.
We have found that similar to saw palmetto, Verbascum xanthophoeniceum extract and its isolated phenylethanoid glycoside verbascoside have no cytotoxicity in human LNCaP prostate epithelial cells, whereas an inhibitory effect on the DHT-induced free and total PSA secretion, a recognized anti-androgen like activity, has been shown in case of both Verbascum xanthophoeniceum extract and pure verbascoside. Furthermore, in the absence of the endogenous androgen DHT, an androgen-like activity in Verbascum xanthophoeniceum is detectable as it is for saw palmetto, suggesting that a mixed androgen-antiandrogen activity is present.
For the first time, Serenoa repens and Verbascum xanthophoeniceum extracts have shown an absence of cytotoxicity and an inhibitory effect on DHT-induced PSA secretion in an in vitro model of human prostate epithelium, whereas the phenylethanoid glycoside verbascoside appeared to explain only part of the Verbascum xanthophoeniceum inhibitory activity on PSA secretion.

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An ethnopharmacological study on Verbascum species: From conventional wound healing use to scientific verification

Abstract

Aim of the study

Material and methods

Results

Conclusion

Graphical abstract

Introduction

Section snippets

Plant material

Results and discussion

Phytochemistry

Journal of Ethnopharmacology

Phytochemistry

Journal of Ethnopharmacology

Journal of Ethnopharmacology

Journal of Ethnopharmacology

Journal of Ethnopharmacology

Journal of Ethnopharmacology

Journal of Ethnopharmacology

Fitoterapia

Antimicrobial and antimalarial activities of some endemic Turkish Verbascum species

FABAD Journal of Pharmaceutical Sciences

Therapy with Medicinal Plants in Turkey (Past and Present)

The effect of cortisone and anabolic steroids on the tensile strength of healing wounds

Annals of Surgery