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The aim of this study was to investigate the ethanol extracts of four medicinal plants, Achillea millefolium L., Hyssopus officinalis L., Equisetum arvense L. and Echinacea purpurea L. and their polyherbal formula, used in traditional medicine for wound healing. The study analyzed their total phenolics content using Folin-Ciocalteu method and identified the main constituents by HPLC. Their antioxidant activity was evaluated by DPPH and ABTS assays and the formula’s capacity to enhance collagen synthesis in L929 fibroblast cell culture was determined by Sircol assay. The results showed that the polyherbal extract had phenolic constituents with pharmacological properties: chlorogenic acid, caffeic acid, luteolin and apigenin. It was showed that the polyherbal formula presented higher antioxidant activity than plant extracts and induced a stimulation of collagen synthesis by fibroblasts, which could contribute to wound strength. In conclusion, the proposed polyherbal formula demonstrated high potential as therapeutic agent in wound healing.
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Studia Universitatis “Vasile Goldiş”, Seria Ştiinţele Vieţii
Vol. 25, issue 1, 2015, pp.41-46
© 2015 Vasile Goldis University Press (
*Correspondence: Dr. Valentina Alexandru, National Institute of R&D for Biological Sciences, 296, Splaiul Independentei, sector 6,
060031, Bucharest, Romania, Tel/fax: +40-21-2200882, E-mail:
Article published: February 2015
Valentina Alexandru1*, Alexandra Gaspar1, Simona Savin1, Agnes Toma1,
Rodica Tatia1, Elvira Gille2
1National Institute of R&D for Biological Sciences, 296, Splaiul Independentei, sector 6, 060031,
Bucharest, Romania
2National Institute of R&D for Biological Sciences, „Stejarul” Biological Research Centre, 6,
Alexandru cel Bun Street, 610004, Piatra Neamt, Romania
ABSTRACT. The aim of this study was to investigate the ethanol extracts of four medicinal plants, Achillea
millefolium L., Hyssopus officinalis L., Equisetum arvense L. and Echinacea purpurea L. and their polyherbal
formula, used in traditional medicine for wound healing. The study analyzed their total phenolics content
using Folin-Ciocalteu method and identified the main constituents by HPLC. Their antioxidant activity was
evaluated by DPPH and ABTS assays and the formula’s capacity to enhance collagen synthesis in L929
fibroblast cell culture was determined by Sircol assay. The results showed that the polyherbal extract had
phenolic constituents with pharmacological properties: chlorogenic acid, caffeic acid, luteolin and apigenin. It
was showed that the polyherbal formula presented higher antioxidant activity than plant extracts and induced
a stimulation of collagen synthesis by fibroblasts, which could contribute to wound strength. In conclusion,
the proposed polyherbal formula demonstrated high potential as therapeutic agent in wound healing.
Keywords: polyherbal formula, wound healing, antioxidant activity, collagen, medicinal plants
Oxidative stress is caused by an imbalance between
the production of reactive oxygen species (ROS) and
the endogenous antioxidant system. The human body
cells are equipped with multiple mechanisms to fight
against ROS and to maintain the cellular redox
homeostasis (Bergendi et al., 1999). When the
antioxidant protection mechanism became unbalanced,
the exogenous antioxidants, such as those from plants,
can help reducing the oxidative damage. The phenolic
compounds (phenolic acids, flavonoids, flavanols,
anthocyanins, etc.) from medicinal plants have been
reported to be potent free radical scavengers (Mathew
et al., 2006). The antioxidant properties of phenolic
compounds have been substantiated by their high
reactivity and potential to chelate metal ions (Rice-
Evans et al., 1997).
In acute and chronic wounds, the expression of
enzymatic antioxidants increased, while their activity
decreased, due to high oxidative stress (James et al.,
2001). Besides, several studies reported that depletion
of non-enzymatic antioxidants was more pronounced in
chronic wounds than in acute wounds (Shukla et al.,
1999; Steiling et al., 1999). Addition of substances
with antioxidant effect was proved to be important in
the successful treatment of skin wounds (Houghton et
al., 2005).
Healing of wounds involves the activity of an
intricate network of blood cells, cytokines and growth
factors, resulting in restoration of normal skin tissue
condition (Clark, 1991). The interest in evaluating the
utility of plant extracts for wound healing has been
increased during the last decade. The importance of
plant secondary metabolites as potential agents that
interfered with various wound repair stages has been
demonstrated, both in vitro and in vivo (Parasanta et
al., 2013; Tsala et al., 2013).
Traditional medicine often used multiple herb
formulae for a wide range of treatments. In skin wound
healing, four medicinal herbs, Achillea millefolium L.
(Compositae), Hyssopus officinalis L. (Labiatae),
Equisetum arvense L. (Equisetaceae) and Echinacea
purpurea L. (Compositae) were used either alone or in
combination with other herbs. These plants contributed
to wound healing and tissue regeneration by multiple
mechanisms, which still need assessment and
validation by scientific studies.
The present study aimed to evaluate, for the first
time, their combination in a particular polyherbal
formula. Its assessment consisted of the identification
and quantification of polyphenolic compounds and the
determination of total antioxidant activity. In order to
support the use of this four-herb formula as a new,
natural product for skin wound healing, it was also
investigated its in vitro effect on collagen secretion by
L929 fibroblast cells in culture.
The plants Equisetum arvense L., Achillea
millefolium L., Hyssopus officinalis L and Echinacea
purpurea L. were collected from Neamt and Suceava
counties, located in the North of Romania. The plant
material was authenticated by prof. dr. Nicolae Stefan
(Botany Department, Faculty of Biology, “Alexandru
Ioan Cuza” University, Iasi). Voucher specimens were
deposited at the Herbarium of Iasi Botanical Garden,
Romania. HPLC-grade gallic acid, chlorogenic acid,
caffeic acid, coumaric acid, ferulic acid, rutoside,
myricetin, luteolin, quercetin, apigenin, acetonitrile and
methanol were purchased from Sigma-Aldrich
Alexandru V., Gaspar A., Savin S., Toma A., Tatia R., Gille E.
Studia Universitatis “Vasile Goldiş”, Seria Ştiinţele Vieţii
Vol. 25, issue 1, 2015, pp. 41-46
© 2015 Vasile Goldis University Press (
(Germany). Butylated hydroxytoluene (BHT), Folin-
Ciocalteu’s phenol reagent, 6-hydroxy-2,5,7,8-
tetramethylchroman-2-carboxylic acid (Trolox), 2,2-
diphenyl-1-picryl-hydrazyl (DPPH) and 2,2’-
azinobis(3-ethylbenzothiazoline-6-sulfonic acid)
diammonium salt (ABTS) and all other chemicals and
solvents of analytical grade were purchased from
Sigma-Aldrich (Germany). The fibroblast cell line
NCTC clone L-929 was purchased from the European
Collection of Cell Cultures (ECACC), minimum
essential medium Eagle (MEM) from Sigma-Aldrich
(Germany) and fetal calf serum (FCS) from Biochrom
AG (Germany). Sircol collagen assay kit was
purchased from Biocolor Ltd. (Newtownabbey, UK).
Extraction procedures
The aerial parts of each plant were air dried, in the
dark and minced using a blender. In order to obtain the
polyherbal extract, dried herbs were mixed as follows:
4 g Equisetum arvense, 3 g Achillea millefolium, 2.5 g
Echinacea purpurea, 0.5 g Hyssopus officinalis. The
mixture (10 g) was extracted in 100 mL ethanol (70 %,
v/v), at room temperature, in the dark, for 10 days.
Then, the polyherbal extract was separated from the
residue by filtration through Whatman No.1 filter paper
and concentrated under vacuum, at 40 °C using a rotary
evaporator (VVMicro, Heidolph, Germany. For cell
culture experiments, the solid residue of the polyherbal
extract, resulted after concentration under vacuum, was
weighed, dissolved in distilled water and sterilized by
filtration through 0.2 µm membrane. On the
experiment day, several extract dilutions were prepared
in the culture medium. Individual plant ethanol extracts
were prepared in the same conditions, in order to be
used as controls.
Total phenolics content assay
Total phenolics content of the herb extracts was
determined using a modified Folin-Ciocalteu method
(Singleton et al., 1999). Briefly, 2.5 mL herb extract
was mixed with 2.5 mL Folin-Ciocalteu reagent and,
after 5 min, 2 mL sodium carbonate (12%, w/w) were
added. The mixture was allowed to stand at room
temperature, for 15 min. The optical density (OD) of
the resulting blue complex was measured at 731 nm
using an UV-Vis spectrophotometer (Jasco V-650,
Japan). Total phenolic content was calculated from the
linear equation of the calibration curve obtained for
chlorogenic acid. The results were expressed as mg
chlorogenic acid equivalents (ChAE)/g dry extract.
DPPH free radical scavenging activity assay
The method is based on scavenging DPPH stable
radical in the presence of hydrogen donor antioxidant,
along with color turn from purple to yellow. We
measured the free radical scavenging activity of each
extract using the method of Hatano et al. (1988) with
some modifications. Briefly, different herb extract
concentrations (10, 25, 50, 100, 250, 500 µg/mL) were
added to DPPH methanol solution (0.25 mM) and each
mixture was incubated in the dark, for 30 min. The OD
was measured at 517 nm against the blank (DPPH
methanol solution), using an UV/VIS
spectrophotometer (Jasco V650, Japan). The inhibition
percentage was calculated using the following formula:
Inhibition (%) = (ODblank-ODsample) / ODblank x100 (1)
The sample concentration that inhibited 50% of
DPPH free radicals (IC50, µg/mL) was calculated from
the graph plotting inhibition percentage against extract
concentration by linear regression analysis. BHT was
used as positive control.
ABTS radical cation scavenging assay
The method is based on the capacity of a sample to
scavenge the ABTS radical cation (ABTS•+), compared
to Trolox as standard antioxidant. We determined the
antioxidant activity of each extract according to the
method of Rice-Evans and Miller (1994). Briefly, 2.5
mL ABTS stock solution (7 mM) in potassium
persulfate (2.45 mM) was mixed with 0.1 mL sample
(herb extract) or standard (Trolox) and 0.4 mL ethanol,
and the mixture was allowed to stand at room
temperature, for 3 min. Then, the OD was recorded at
731 nm against the blank, containing all reagents
except the tested extract, at an UV/VIS
spectrophotometer (Jasco V 650). The results were
expressed as Trolox equivalents antioxidant capacity
(TEAC) calculated using the formula:
TEAC (μM Trolox equivalents/g dry weight) = CTrolox x
f x (ODsample ODblank) / (ODTrolox ODblank) (2)
where Ctrolox is Trolox concentration and f is the
sample dilution factor.
HPLC analysis
The separation, identification and quantification of
phenolic compounds from polyherbal extract were
performed by HPLC, using an Agilent 1200 system
(Agilent, USA) equipped with diode array detector and
Eclipse XDB-C18 (150 x 4.6 mm i.d.; 5 µm particles)
chromatographic column, after injection of 10 µl
sample. The mobile phase used for phenolics
separation was a mixture of phase A (2 mM sodium
acetate buffer, pH 3.05) and phase B (acetonitrile), in
linear gradient mode, as follows: 0-30 min, 2-20% B in
A; 30-40 min, 20-30% B in A; 40-50 min, 30% B in A;
50-60 min, 30%-2% B in A. The flow rate was 1
mL/min. Chromatograms were recorded at a
wavelength of 260 nm for phenolic acids and 320 nm
for flavonoids. The constituents present in the
polyherbal extract were identified by comparing the
recorded UV profile and their retention times with
those obtained for a mixture of known standards of
phenolic acids (gallic acid, chlorogenic acid, caffeic
acid, p-coumaric acid, ferulic acid) and flavonoids
(rutoside, myricetin, luteolin, quercetin, apigenin). In
order to calculate the content of each polyphenol
identified in the polyherbal extract, calibration curves
for standards were built as five-point plots, in the range
of 0.976 15.625 µg/mL.
Soluble collagen assay
Mouse fibroblasts cell culture (NCTC cell line
clone L929) was used to study the effect of polyherbal
extract on collagen secretion. Cells were seeded in the
wells of 24-well culture plate, at a density of 5x104
cells/well in MEM supplemented with10% FCS.
Phenolic content, antioxidant activity and effect on collagen synthesis of a traditional wound healing polyherbal formula
Studia Universitatis “Vasile Goldiş”, Seria Ştiinţele Vieţii
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After 24 h of incubation in standard conditions, the
cells adhered to plastic and the culture medium was
changed with MEM supplemented with 5% FCS,
containing different concentrations of polyherbal
extract (35-140 µg/mL). The plates were incubated at
37 ºC, in humidified 5% CO2 air atmosphere, for 48 h
and 72 h, respectively. The control group consisted of
untreated cells cultivated in MEM with 5% FCS.
Collagen secretion in the culture medium was
determined using Sircol collagen assay kit, according
to manufacturer’s instructions. Briefly, the harvested
culture media were centrifuged at 1,500 rpm, for 4 min
and, then, 100 μl supernatant was mixed with 1 mL
Sircol dye, for 30 min. The mixture was centrifuged at
10,000 rpm, for 5 min to precipitate the collagen-dye
complex. Then, the pellets were dissolved in 1 mL
alkali reagent and vortexed. The OD of the solution
was read at 540 nm using Sunrise microplate reader
(Tecan, Austria).
Statistical analysis
All chemical analyses were run in triplicate and
three cell culture independent experiments were
performed in three replicates. Data were reported as
mean ± standard deviation (SD). Pair comparison of
control and each sample was carried out by t-test.
Significant statistical differences were considered at p
< 0.05.
Total phenolics content and antioxidant
The results of total phenolics content in E. arvense,
H. officinalis, A. millefolium, E. purpurea ethanolic
extracts and their polyherbal formula extract are
showed in Fig. 1. The amount of total phenolics in
plant extracts varied from 8.95 mg ChAE/g dry extract
for E. purpurea, to 12.43 mg ChAE/g dry extract for A.
millefolium. Significant (p < 0.05) higher phenolic
compounds level was detected in the polyherbal extract
(14.42 mg ChAE/g dry extract), compared to each
plant extract.
Fig. 1 Total phenolics content in plant extracts and polyherbal (PH) extract. *p < 0.05, compared to each plant extract
In order to determine the antioxidant activity of the
polyherbal extract, in comparison to individual plant
extracts, two complementary test systems have been
applied, DPPH and ABTS assays. The results of DPPH
assay showed the extract concentration that resulted in
50% DPPH free radical inhibition (IC50) (Fig. 2).
Fig. 2 Radical scavenging activities of plant extracts and polyherbal (PH) extract on DPPH radical. Results were
expressed as IC50 (mean ± SD). BHT was used as standard reference. *p<0.05, compared to BHT; #p<0.05, compared
to plant extracts.
Alexandru V., Gaspar A., Savin S., Toma A., Tatia R., Gille E.
Studia Universitatis “Vasile Goldiş”, Seria Ştiinţele Vieţii
Vol. 25, issue 1, 2015, pp. 41-46
© 2015 Vasile Goldis University Press (
IC50 values decreased in the folowing order: H.
officinalis >E. arvense >E. purpurea >A. millefolium >
polyherbal extract. Therefore, the polyherbal extract
presented a significant (p < 0.05) higher radical
scavenging activity than individual plant extracts, but
significant (p < 0.05) lower than BHT, a well-known
synthetic antioxidant.
The antioxidant activities of plant extracts
evaluated by ABTS assay varied from 98.13 µM
Trolox equivalents/g dry extract for E. arvense to
176.19 µM Trolox equivalents/g dry extract for A.
millefolium (Table 1). The polyherbal extract had the
highest antioxidant activity (254.88 μM Trolox
equivalents/g dry extract). This result was in
accordance with that obtained by DPPH assay and
correlated with its phenolic compounds content.
Table 1.
Antioxidant activities of plant extracts and polyherbal formula extract evaluated by ABTS assay
Plant species
ABTS assay
(μM Trolox equivalents/g dry weight)
Hyssopus officinalis
171.73 ± 4.54
Echinacea purpurea
168.22 ± 8.48
Achillea millefolium
176.19 ± 4.96
Equisetum arvense
98.13 ± 3.84
Polyherbal extract
254.88 ± 12.67*
*p < 0.05, compared to each plant extract
All these data showed that the polyherbal extract
exhibited higher phenolics content and antioxidant
capacity, compared to its component extracts of A.
millefolium, E. purpurea, E. arvense and H. officinalis.
As a result, the polyherbal extract was tested in
subsequent analyzes.
Chemical composition of the polyherbal
The established HPLC method was applied as
analytic approach to determine the major compounds
of the polyherbal extract. The recorded HPLC profile
presented nine main peaks, at 1.312, 6.352, 18.616,
21.899, 24.819, 25.169, 29.190, 29.541 and 45.067 min
(Fig. 3).
Fig. 3 Chromatographic profile of polyphenolic constituents from the polyherbal extract recorded by HPLC
A mixture of known pure compounds was also
chromatographed and used as external standards of
phenolic acids (gallic acid, chlorogenic acid, caffeic
acid, coumaric acid and ferulic acid) and flavonoids
(rutoside, myricetin, luteolin, quercetin and apigenin).
The values of retention time for these standards and
their calibration curves parameters are presented in
Table 2. Table 2.
Analytical results of calibration curves of ten polyphenolic compounds used as standards in HPLC analysis
Retention time
Regression equation of
the calibration curvea
Correlation factor
Gallic acid
y = 28.963x + 41.860
Chlorogenic acid
y = 14.856x + 19.038
Caffeic acid
y = 24.325x + 39.374
Coumaric acid
y = 19.319x + 33.477
Ferulic acid
y = 24.906x + 42.230
y = 14.240x + 20.226
y = 33.706x + 99.057
y = 58.234x + 179.806
y = 45.482x + 139.321
y = 33.958x + 121.724
aThe calibration curves were plotted in linear regression analysis of the integrated peak area (y) versus concentration (x)
Phenolic content, antioxidant activity and effect on collagen synthesis of a traditional wound healing polyherbal formula
Studia Universitatis “Vasile Goldiş”, Seria Ştiinţele Vieţii
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In order to determine the content of the identified
compounds in polyherbal extract, quantitative
calculations were performed by peak area integration.
The results of HPLC analysis showed that the
polyherbal extract presented high levels of chlorogenic
acid (1.226 mg/g dry extract), rutoside (1.605 mg/g dry
extract), apigenin (0.982 mg/g dry extract) and luteolin
(0.692 mg/g dry extract) (Table 3). Low levels of
caffeic acid, coumaric acid and quercetin were
quantified in the polyherbal extract (Table 3).
Table 3.
Content of phenolic acid and flavonoid compounds
identified in the polyherbal extract
Gallic acid
Chlorogenic acid
Caffeic acid
Coumaric acid
Ferulic acid
ND - not detected
Previous studies showed that these phenolic
compounds presented several pharmacological
properties and exerted anti-inflammatory, antioxidant,
antiviral, antibacterial and vulnerary activities (Fuchs
et al., 1993; Morishita et al., 2001; Song et al., 2008;
Lopez-Lazaro, 2009; Kostyuk et al., 2010).
Effect of the polyherbal extract on collagen
Wound healing is a fundamental response to tissue
injury. The present knowledge described three phases
of this process: inflammatory phase, proliferative phase
and remodelling phase. In the proliferative phase,
fibroblasts produced a variety of substances, essential
for wound repair, including glycosaminoglycans and
collagen (Madden et al., 1968). In the remodeling
phase, new collagen was formed and tissue tensile
strength was increased due to intermolecular cross-
linking of collagen, via vitamin C-dependent
hydroxylation (Prockop et al., 1979; Stadelmann et al.,
1998). In Fig. 4 is presented the influence of polyherbal
extract on the synthesis of soluble collagen, after
cultivation in different concentrations with L929
fibroblast cells.
Fig. 4 Determination of collagen secretion by L929 fibroblast cells incubated with different concentrations of polyherbal
extract, for 48 h and 72 h, using Sircol assay. Three independent experiments were performed with three replicates for
each sample. Values are mean ± SD. *p < 0.05, compared to control (untreated cells).
The results showed a significantly (p < 0.05)
increase of collagen synthesis in the culture medium of
fibroblasts treated with 70 and 140 µg/mL polyherbal
extract, after 48 h and 72 h of cultivation. It was
observed that the collagen synthesis was almost 2 times
higher in cultures treated with 140 µg/mL polyherbal
extract, for 72 h, compared to the value obtained in the
control group (0 µg/mL polyherbal extract).
Previous studies reported that natural polyphenols
presented reducing properties, protection of
intracellular lipids from oxidation and influenced
collagen synthesis (Mucha et al., 2013). Our
experimental data suggested that increased collagen
synthesis in L929 fibroblast cell culture was correlated
to high phenolics content and, by default, with the
antioxidant activity of the polyherbal extract.
These results support the traditional use of this
four-herb formula for wound care. The polyherbal
extract had higher total phenolics content and
antioxidant activity, compared to individual plant
extracts. It was showed its in vitro capacity to stimulate
collagen synthesis in a culture of fibroblast cells.
Therefore, this combination of plant extracts may be
useful as therapeutic agent in wound healing. Future
studies could be performed, in order to find out its
unexplored efficacy and high potential as a source of
natural health care products.
Alexandru V., Gaspar A., Savin S., Toma A., Tatia R., Gille E.
Studia Universitatis “Vasile Goldiş”, Seria Ştiinţele Vieţii
Vol. 25, issue 1, 2015, pp. 41-46
© 2015 Vasile Goldis University Press (
This study was supported by the Executive Unit for
Financing Higher Education, Research, Development
and Innovation (UEFISCDI), Project No. 62071 and
Romanian Project BIODIV No. 102.
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... Kaempferol 3-O-sophoroside and kaempferol 3-O-sophoroside-7-O-glucoside, isolated from E. giganteum , are reported to be used as potent pharmacological agents in treating gastrointestinal diseases, oxidative stress, and histologic changes (Campos-Vidal et al., 2022). Apigenin 5-O-glucoside, isolated from E. arvense (Zhang et al., 2015), was revealed as an effective Caffeic acid, p-coumaric acid (Uslu et al., 2013;Alexandru et al., 2015); vanillic acid (Uslu et al., 2013); ferulic acid ( (Syrchina et al., 1980) ...
... Benzoic acid, isolated from the stems of E. arvense, E. fluviatile, and E. telmateia (Hiraga et al., 1997), is a commonly used preservative in acidic foods, beverages and also a widely used compound in pharmaceutical industry (Zu et al., 2017). Gallic acid, chlorogenic acid, myricetin (Alexandru et al., 2015), trans-caffeoyl-3-β-glucoside (Park and Tomohiko, 2011), (6R,7aS)-5,6,7,7a-tetrahydro-6-hydroxy-4, 4,7a-trimethyl benzofuran-2(4H)-one , and a phenylpropanoid, rosmarinic acid (Alves et al., 2016) were isolated from the aerial parts of E. hyemale and these compounds have antioxidant, anti-inflammatory, anticancer, antidiabetic, osteoporosis protection, gastrointestinal, neuropsychological, cardiovascular disorders, anti-obesity, hepatoprotective, and antineoplastic properties (Naveed et al., 2018;Kahkeshani et al., 2019;Imran et al., 2021;Guan et al., 2022). ...
... Methanolic extracts of the whole plant parts of E. arvense showed 90% DPPH inhibition, and 70% nitric oxide inhibition and 60% H 2 O 2 inhibition (Amit et al., 2013). Similarly, a number of studies have found that the whole plant parts and aerial parts of E. arvense have significant antioxidant effects (Jia-Gua et al., 2006;Stajner et al., 2006Stajner et al., , 2009Oka et al., 2007;Milutinovic et al., 2014;Cetojevic-Simin et al., 2010;Uslu et al., 2013;Alexandru et al., 2015;Huh and Han, 2015). Aqueous extracts and fractions of E. arvense aerial parts showed notable DPPH and hydroxyl free radical scavenging effects with an EC 50 of 1.52 ± 0.07 mg/mL (ethyl acetate fraction), 0.64 ± 0.03 mg/mL (n-butanol fraction), and 2.27 ± 0.11 mg/mL (aqueous extracts) . ...
Introduction: The genus Equisetum (Equisetaceae) is cosmopolitan in distribution, with 41 recognized species. Several species of Equisetum are widely used in treating genitourinary and related diseases, inflammatory and rheumatic problems, hypertension, and wound healing in traditional medicine practices worldwide. This review intends to present information on the traditional uses, phytochemical components, pharmacological activities, and toxicity of Equisetum spp. and to analyze the new insights for further study. Methods: Relevant literature has been scanned and collected via various electronic repositories, including PubMed, Science Direct, Google Scholar, Springer Connect, and Science Online, from 1960 to 2022. Results: Sixteen Equisetum spp. were documented as widely used in traditional medicine practices by different ethnic groups throughout the world. A total of 229 chemical compounds were identified from Equisetum spp. with the major group of constituents being flavonol glycosides and flavonoids. The crude extracts and phytochemicals of Equisetum spp. exhibited significant antioxidant, antimicrobial, anti-inflammatory, antiulcerogenic, antidiabetic, hepatoprotective, and diuretic properties. A wide range of studies have also demonstrated the safety of Equisetum spp. Conclusion: The reported pharmacological properties of Equisetum spp. support its use in traditional medicine, though there are gaps in understanding the traditional usage of these plants for clinical experiments. The documented information revealed that the genus is not only a great herbal remedy but also has several bioactives with the potential to be discovered as novel drugs. Detailed scientific investigation is still needed to fully understand the efficacy of this genus; hence, very few Equisetum spp. were studied in detail for phytochemical and pharmacological investigation. Moreover, its bioactives, structure-activity connection, in vivo activity, and associated mechanism of action ought to be explored further.
... Many authors in this context have reported the phytochemical composition of the poly-herbal compounds in relation to the wound healing potentials [7,15,17,22,25,29,35]. In this connection, many authors have reported secondary metabolites having an antioxidant nature, such as flavonoids and active phenol compounds present to be responsible for the wound healing process [8,18,24,29,71]. ...
... Many studies have suggested various targets for these active compounds that ultimately help to heal. These are mediated by multiple cascades, which includes mitogenic pathways [22,24,25,50], extracellular matrix synthesis pathway [11], free radical scavenging pathway [13,29e31,33,36,41], atherosclerosis pathway [19] and anti-inflammatory pathways [48,49,51]. It is therefore evident that the majorly studied pathway from the aforementioned is the free radical scavenging pathway. ...
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Background: The disruptions in skin integrity contribute to its disorientation, and tissue annihilations result in acute or chronic wound formation. Polyherbal formulations are widely used in traditional systems of mecdicine like ayurveda for wound healing. The combination of these traditional therapies with clinical therapies has helped in the development of various wound-healing products. Method: In this systematic review, the therapeutic potency of several polyherbal formulations from different medicinal floras is summed together in response to their impact on wound healing. The literature search was performed on Pubmed, Scopus, and ScienceDirect databases between 2010-2020. PRISMA methodology was applied to extract relevant information about polyherbal formulations. Result: A total of 54 articles were selected under all themes for the data extraction as per the PRISMA guidelines. These 54 articles have high-quality scores ≥3. Forty-three records were used for the narrative analysis, while nine records were used for the critical analysis in the narrative review. Further, theme-wise key data sets were screened from the selected literature and summarized in a tabular form. Bibliometric analysis of the Scopus database has also drawn attention to limited academic literature showcasing randomized clinical trials in the current subject. Most of these polyherbal formulations are tested in laboratory-scale studies, thus portraying further research options. Conclusion: Polyherbal formulations are effective in promoting the wound-healing process. They can stimulate a variety of physiological functions that accelerates the process of healing. These formulations merit further investigation in clinical trials, and production up scaling will aid in the creation of a new horizon of polyherbal wound healing products.
... However, if the endogenous antioxidant system fails to balance the production and degradation of ROS, the excessive levels of ROS create a condition of oxidative stress, which activates pro-apoptotic proteins, generating toxic effects on cells, causing inflammation and delay in the healing process. Exogenous antioxidants such as those found in plant metabolites are an alternative to reduce oxidative damage, having a beneficial effect on wound healing [32,[68][69][70][71][72]. In previous studies carried out in our laboratory, the antioxidant capacity (SA 50 ) of ChEEP was determined, and the extract showed an SA 50 = 15.75 µg/mL [27], in agreement with the antioxidant activity index is considered a very strong antioxidant activity [73]. ...
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Skin wound healing is a complex biochemical process of tissue repair and remodeling in response to injury. Currently, the drugs used to improve the healing process are inaccessible to the population, are costly, and have side effects, making the search for new treatment alternatives necessary. Propolis is a natural product produced by bees that is widely recognized and used in folk medicine for its multiple biomedical activities. However, therapeutic information regarding Mexican propolis is limited. This study aimed to evaluate the wound-healing effect of the Chihuahua ethanolic extract of propolis (ChEEP). Macroscopic and histological analyses were performed using a mouse wound-healing model. The topic acute toxicity assay showed that propolis at 10% w/v had no toxic effects. ChEEP has antibacterial activity against the Gram-positive bacteria Staphylococcus aureus and Staphylococcus epidermidis. Moreover, it exhibited good anti-inflammatory activity evaluated through mouse ear edema induced by 12-O-tetradeca-noylphorbol-13-acetate (TPA). A full-thickness incision lesion was created in mice and treated topically with 10% ChEEP. At Day 14 post-treatment, it was observed that propolis increased wound contraction and reduced healing time and wound length; furthermore, propolis increased the tensile strength of the wound, as determined with the tensiometric method, and promoted the formation of type I collagen at the site of injury, as evaluated with Herovici stain. These findings suggest that the topical administration of ChEEP can improve skin wound healing, probably due to the synergistic effect of its components, mainly polyphenols, in different steps of the wound-healing process. It should be noted this is the first time that the wound-healing activity of a Mexican propolis has been evaluated.
... In clinical trials, topical phytotherapy combining E. arvense extracts with other herbal extracts showed benefits to brittle nail disorder. Ointment containing E. arvense (3%) showed wound-healing effects [28]. Other clinical trials reported the efficacies of E. arvense for benign prostate hyperplasia and chronic musculoskeletal pain [29]. ...
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Trends in skin and hair treatments focus on natural products due to undesired effects of chemically synthetic ingredients. This study aims to investigate the cosmeceutical effects of Equisetum debile (horsetail) extracts relating to anti-hyperpigmentation via tyrosinase, anti-wrinkle formation via matrix metalloproteinases (MMPs), and anti-androgenic alopecia via 5α-reductase. Ethanolic extracts were sequentially partitioned into semi-purified fractions hexane, dichloromethane, ethyl acetate, methanol, and methanol insoluble residue. The ethyl acetate fraction possessed the highest total phenolic content (39.24 ± 0.72 mg gallic acid/g), the strongest anti-tyrosinase activities (583.33 ± 23.59 mg kojic acid/g), and potent collagenase inhibitions (IC50 MMP-1 and MMP-2 of 0.82 ± 0.09 and 0.94 ± 0.11 mg/mL, respectively). All extracts showed considerable inhibitions of 5α-reductase ranging from 44.59 ± 0.40 to 83.07 ± 3.46% with the strongest activity from the dichloromethane fraction (1.48 ± 0.06 mg finasteride/g). In conclusion, E. debile extracts exhibit cosmeceutical potentials. This study suggests that the E. debile ethyl acetate fraction could be used as a promising ingredient to organically treat hyperpigmentation and delay the skin aging process. In addition, compared to the current recommended intake of finasteride (1 mg/day) for androgenic alopecia, the dichloromethane fraction is proposed as an alternative source to naturally remediate hair loss.
... Different botanical drug formulations of C. arvensis, L. stoechas, and H. italicum exhibited in vitro regenerative properties as these increase the fibroblast proliferation and migration [93]. A botanical drug consisting of Achillea millefolium L., Hyssopus officinalis L., Equisetum arvense L. and Echinacea purpurea L. showed increased collagen synthesis activity by mouse fibroblasts in vitro, thereby suggestive of wound healing capacity [94]. Another botanical drug consisting of Vitex negundo L., Emblica officinalis Gaertn, and Tridax procumbens L. had demonstrated wound healing activity by rapid movements of keratinocytes and fibroblasts to the wound site, increased rates of skin regenerations, wound contraction, and collagen synthesis [95]. ...
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Skin, the largest organ of the body, plays a vital role in protecting inner organs. Skin stem cells (SSCs) comprise a group of cells responsible for multiplication and replacement of damaged and non-functional skin cells; thereby help maintain homeostasis of skin functions. SSCs and differentiated cells of the skin such as melanocytes and keratinocytes, have a plethora of applications in regenerative medicine. However, as SSCs reside in small populations in specific niches in the skin, use of external stimulants for cell proliferation in vitro and in vivo is vital. Synthetic and recombinant stimulants though available, pose many challenges due to their exorbitant prices, toxicity issues and side effects. Alternatively, time tested traditional medicine preparations such as polyherbal formulations are widely tested as effective natural stimulants, to mainly stimulate proliferation, and melanogenesis/prevention of melanogenesis of both SSCs and cells of skin origin. Complex, multiple targets, synergistic bioactivities of the phytochemical constituents of herbal preparations amply justify these as natural stimulants. The use of these formulations in clinical applications such as in skin regeneration for burn wounds, wound healing acceleration, enhancement or decrease of melanin pigmentations will be in great demand. Although much multidisciplinary research is being conducted on the use of herbal formulas as stem cell stimulants, very few related clinical trials are yet registered with the NIH clinical trial registry. Therefore, identification/ discovery, in depth investigations culminating in clinical trials, as well as standardization and commercialization of such natural stimulants must be promoted, ensuring the sustainable use of medicinal plants. Graphical Abstract
... Like the current data. Many previous studies confirmed that E. arvense extract has high amounts of total phenols, total flavonoids, and total antioxidants [63][64][65][66] ...
Methotrexate (MTX) is a cytotoxic drug used to treat a wide range of cancers and non-cancerous conditions. However, it can cause unfavorable acute toxic effects in several organs, including the testis. Equisetum arvense L. (E. arvense) extract is effective in counteracting oxidative stress-related disorders. This study assessed the preventive effect of E. arvense extract against MTX-induced testicular toxicity. Gas chromatography-mass spectroscopy (GC-MS) was used to analyze the active constituents of E. arvense extract. Testicular toxicity was induced via MTX injection (0.5 mg/kg/ twice a week for 4 weeks). Forty male albino rats were divided into 4 groups: I- control (Cont); II: MTX; III: E. arvense (500 mg/kg/daily for 10 weeks); and IV: E. arvense + MTX. E. arvense main active constituents were squalane (15%), ascorbic acid per methyl (9.55%), phytol (8.69%), 2-pyrroline 1,2-dimethyl (8.63%), and octacosane (8.23%). Treatment of MTX injected rats with E. arvense produced a significant rise in body weight, serum testosterone and luteinizing hormone. E. arvense significantly increased the sperm counts, viability, and motility relative to the MTX injected rats. The levels of testicular oxidative stress and inflammation significantly reduced in the MTX rats treated with E. arvense. Furthermore, E. arvense markedly improved the testicular tissue and seminiferous tubules’ pathologic features in MTX-treated rats. E. arvense significantly decreased lipid peroxidation products, interleukin-1 beta, tumor necrosis factor-alpha while increasing superoxide dismutase levels. E. arvense prevented MTX-induced testicular damage via anti-inflammatory and antioxidant activities.
... The ability of CA to stimulate the wound healing process was already investigated in the past, indicating that this bioactive molecule presented not only low cytotoxicity toward fibroblast cells [136] but also the tendency to increase their proliferation towards the repair of injured tissues [137]. Alexandru and co-workers [138] demonstrated that CA extracted from four medical plants stimulated collagen synthesis by fibroblast cells, confirming its high potential as a therapeutic agent in wound healing. CA also represents one of the active ingredients of the Polygonum aviculare extract, which was observed to promote the migration of keratinocyte and fibroblast cells, efficiently accelerating the epithelialization process of wounds in a murine model [77]. ...
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Alterations of skin homeostasis are widely diffused in our everyday life both due to accidental injuries, such as wounds and burns, and physiological conditions, such as late-stage diabetes, dermatitis, or psoriasis. These events are locally characterized by an intense inflammatory response, a high generation of harmful free radicals, or an impairment in the immune response regulation, which can profoundly change the skin tissue’ repair process, vulnerability, and functionality. Moreover, diabetes diffusion, antibiotic resistance, and abuse of aggressive soaps and disinfectants following the COVID-19 emergency could be causes for the future spreading of skin disorders. In the last years, hydroxycinnamic acids and derivatives have been investigated and applied in several research fields for their anti-oxidant, anti-inflammatory, and anti-bacterial activities. First, in this study, we give an overview of these natural molecules’ current source and applications. Afterwards, we review their potential role as valid alternatives to the current therapies, supporting the management and rebalancing of skin disorders and diseases at different levels. Also, we will introduce the recent advances in the design of biomaterials loaded with these phenolic compounds, specifically suitable for skin disorders treatments. Lastly, we will suggest future perspectives for introducing hydroxycinnamic acids and derivatives in treating skin disorders.
... In the present study, another compound extracted from the Verbascum Thapsus was phenol. Plant extracts that have high phenolic components, due to their antioxidant, antibacterial, and anti-inflammatory activity, have a high potential for rapid repair of damaged skin tissues; Besides, through stimulating collagen synthesis by fibroblasts, they can help increase wound consistency [41]. In the study of Yadav (2018), the use of phenol-rich topical ointment potentially accelerated the hydroxyproline content and collagen synthesis in the wound [42]. ...
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Background The pain and discomfort caused by episiotomy affect the quality of life of the mothers, so rapid and complete repair of the episiotomy is very important. Due to the effective ingredients of Verbascum Thapsus , it has been used since ancient times to treat wounds. Therefore, this study aimed to evaluate the effect of Verbascum Thapsus on episiotomy wound healing. Methods The study was designed as a randomized, double-blind, controlled clinical trial. Ninety-three primiparous women who were referred to Fatemeh Zahra Hospital in Saveh in 2015 were randomly divided into two groups of intervention ( Verbascum Thapsus ) and control (placebo). Both groups covered the episiotomy wound twice a day for 10 days with 2 cm of prescribed creams. Wound healing was assessed using the REEDA scale before the intervention and on days 1,3 and 10 after the intervention. Results Before the intervention, there was no statistically significant difference in terms of demographic characteristics, obstetrics, and REEDA scores between the two groups ( p < 0.05). The mean scores of REEDA on days 1 and 3 in the intervention group were better than the control group but were not statistically significant. However, on the tenth day after the intervention, the mean scores of REEDA were significantly better in the Verbascum group than the placebo ( p = 0.01). Conclusions According to the results of this study, it seems that Verbascum Thapsus is effective in repairing episiotomy wounds. The researchers hope that the results of this study can provide clinical evidence for the use of this herbal medicine in the wound healing process. Trial registration This study was registered in the Iranian Registry of Clinical Trials (IRCT) with the code “ IRCT201404073106N15 ” on 02/12/2015.
Este estudo foi conduzido para verificar o efeito do extrato aquoso e da pomada à base de casca de Caryocar brasiliense sobre a retração de feridas em coelhos. Foram utilizados 12 coelhos Nova Zelândia Branco divididos em dois grupos e quatro lesões foram produzidas na região dorsal de cada animal. As lesões à direita foram tratadas com extrato aquoso (grupo 1) ou com a pomada (grupo 2) e as lesões à esquerda foram tratadas com solução salina 0,9% (grupo controle). Os níveis de colágeno e fibroblastos foram menores (P<0,05) em lesões tratadas com extrato aquoso, comparado com o grupo controle. Aos 7 e 14 dias após o procedimento, a retração das lesões era maior (P<0,05) quando tratadas com extrato aquoso e nas lesões tratadas com a pomada, a melhora ocorreu apenas no 7º dia, comparado com o tratamento controle. Concluiu-se que o extrato aquoso de casca de Caryocar brasiliense melhorou a retração de feridas por um período de tempo maior do que a pomada.
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Background Equisetum ramosissimum Desf. ( E. ramosissimum ) is a widely used traditional medicinal plant to treat urinary tract infections (UTIs) by ethnic people throughout the world. The utility of the plant in treating urinary-related disorders was evaluated against selected pathogenic bacteria which has major role in causing UTIs. Hence, the present study executed to extract phytochemicals like total phenolics and flavonoids, chemical profiling by GC–MS analysis and to test their antioxidant activity from stem extracts of E. ramosissimum . The extraction process was directed by petroleum ether, chloroform, ethyl acetate, methanol, and aqueous solvents. Results The GC–MS analysis yielded 24 phytoconstituents with linoleic acid, palmitic acid, nonacosane, hexahydrofarnesyl acetone, and octacosane as major compounds. Methanolic extract yielded maximum amount of phenolics (TPC) and flavonoids (TFC) with 600.02 ± 0.22 mg GAE/g and 631.38 ± 0.69 mg QE/g, respectively. Methanolic extract also exhibited notable free radical scavenging activity with an IC 50 of 123.89 ± 0.73, 150.10 ± 1.02, 146.01 ± 0.54, and 63.73 ± 6.12 µg/mL for DPPH, FRAP, ABTS, and O 2 ⁻ assays, respectively. The minimum inhibitory concentration (MIC) required to inhibit the growth of tested pathogenic bacteria was observed in aqueous and methanolic extracts with the value being 31.25 µg/mL against R. equi and V. cholerae . As like, methanolic and petroleum ether extracts efficiently inhibited the growth of B. subtilis with the MIC of 31.25 µg/mL. Conclusion It was concluded that the notable effect of methanolic and aqueous extracts against the uropathogenic bacteria reported in this study supported the traditional uses of this plant in treating UTIs. The results acquired from this investigation revealed that E. ramosissimum stem extract might be considered as an interesting candidate in the development of antibacterial agent against UTIs coupled with antioxidant properties.
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p>Data generated through systematic investigation, carried out on the evaluation of phyto-extracts on wound healing research during the last 20 years have been compiled. About 450 plant species having wound healing properties have been identified. The present knowledge of the wound healing process comprise coagulation, inflammation, proliferation, formation and accumulation of fibrous tissues, collagen deposition, epithelialization, contraction of wound with formation of granulation tissues, remodeling and maturation. The constituents of the plant extracts modulate one or more of the above stages. It was the endeavor to identify the active constituents responsible for antimicrobial activity, free radical scavenging properties, stimulators of enhanced collagen production and/or angiogenesis promoters with identification of lead scaffold chemical structures. Multiple phytochemicals concentrated and blended in optimal concentrations, are expected to be available in future years to carry out multi-tasking efforts in wound healing as more knowledge about the properties of the key constituents are unveiled. This article is open to POST-PUBLICATION REVIEW . Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.</p
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A wound is a disruption of the normal anatomical structure and function of a tissue. Wounds cure in an orderly and timely repair process which is characterized by thr-ee dynamic and interactive phases: inflammation, proliferation and the remodeling. The present review was designed to elaborate the cellular and molecular targets for plant secondary metabolites that target the various aspects of wound repair process. The common mechanism of action of natural products established through in vitro and animal studies include direct action on skin cells regeneration, increase in con-nective tissue deposition, antioxidant activity, inflammatory cells activity and mod-ulation of cytokine and growth factor production and/or function. All these demon-strated pharmacological effects could be exploited to overcome an acute or path-ological wound healing conditions. The therapeutic potential of various chemical classes of natural products that act through one or multiple targets are discussed.
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Inflammation is a key event in the skin normally occurring in response to the constant exposure to environmental and endogenous stimuli as well as to accidental damage. It also plays a central role in the pathogenesis of major cutaneous pathologies, ultimately resulting in skin carcinogenesis. As the acute mild inflammatory process is mainly adaptive in nature, chronic inflammation is a multi-factorial and complex noxious process, extremely difficult to combat. The wealth of data documenting the involvement of redox-dependent regulatory and damaging processes in the skin inflammation has prompted research on a steadily growing number of plant-derived active substances, mainly polyphenols, and selected principally on the basis of their free radical scavenging and antioxidant properties. In spite of the wide recognition of their anti-inflammatory efficacy in vitro, the clinical use for the prevention and treatment of major skin inflammatory conditions is in most cases yet to be conclusively proven. The complex nature of the cutaneous inflammatory process involves oxygen (ROS), nitrogen (RNS) and lipid reactive species, but is also driven by other mechanisms highlighted in these recent years, related to the regulation of gene expression, and to metabolic and signaling pathways that are ROS/RNS-independent. The screening of the enormous array of plant secondary metabolites, first of all polyphenols, for new effective and safe anti-inflammatory agents should be rather directed towards molecules targeting specific inflammatory pathways recognized to be active in the peculiar skin compartment.
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This study was carried out to investigate the wound healing effect of caffeic acid in skin-incised mice. Caffeic acid showed significant effects on anti-inflammatory activity and wound healing, such as myeloperoxidase activity, lipid peroxidation, phospholipase A(2) activity and collagen-like polymer synthesis, in incised-wound tissue. On the other hand, it significantly stimulated collagen-like polymer synthesis in NIH 3T3 fibroblast cells, while inhibited both silica-induced reactive oxygen species generation and melittin-induced arachidonic acid release and PGE(2) production in Raw 264.7 cells, and histamine release in RBL 2H3 cells stimulated by melittin or arachidonic acid. Therefore, caffeic acid appears to have a potent antioxidant and anti-inflammatory effect in cell culture system, which may be related to wound healing in skin-incised mice.
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Epidemiological evidence suggests that flavonoids may play an important role in the decreased risk of chronic diseases associated with a diet rich in plant-derived foods. Flavonoids are also common constituents of plants used in traditional medicine to treat a wide range of diseases. The purpose of this article is to summarize the distribution and biological activities of one of the most common flavonoids: luteolin. This flavonoid and its glycosides are widely distributed in the plant kingdom; they are present in many plant families and have been identified in Bryophyta, Pteridophyta, Pinophyta and Magnoliophyta. Dietary sources of luteolin include, for instance, carrots, peppers, celery, olive oil, peppermint, thyme, rosemary and oregano. Preclinical studies have shown that this flavone possesses a variety of pharmacological activities, including antioxidant, anti-inflammatory, antimicrobial and anticancer activities. The ability of luteolin to inhibit angiogenesis, to induce apoptosis, to prevent carcinogenesis in animal models, to reduce tumor growth in vivo and to sensitize tumor cells to the cytotoxic effects of some anticancer drugs suggests that this flavonoid has cancer chemopreventive and chemotherapeutic potential. Modulation of ROS levels, inhibition of topoisomerases I and II, reduction of NF-kappaB and AP-1 activity, stabilization of p53, and inhibition of PI3K, STAT3, IGF1R and HER2 are possible mechanisms involved in the biological activities of luteolin.
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Genetic defects at the level of transscription and translation may explain why very little type II procollagen is synthesized by patients with type IV of Ehlers-Danlos syndrome. Similary defect synthesis of type I collagen may explain the decrease in the ratio of type I collagen to type III collagen in osteogenesis imperfecta. The synthesized type I collagen in this disorder is unusually sensitive to pepsin, possibly due to an amino acid substitution in the pro-alpha chain of type I procollagen. Defects in intracellular processing include a decrease in lysyl hydroxylase in the type VI variant of Ehlers-Danlos syndrome. Defects in extracellular processing include a decrease in procollagen aminoprotease seen in dermatosparexis and in Ehlers-Danlos syndrome type VII. A deficiency in the enzyme lysyl oxidase is seen in type V Ehlers-Danlos syndrome. A defective cross-linking of collagen may also explain the tissue changes in Marfan's syndrome and in homocysteinuria. Mechanisms for gene selection of collagen may be influenced by environmental factors such as lysosomal enzymes which can change the collagen synthesized by chondrocytes from type II to type I. Scurvy is a classical example of how collagen biosynthesis can be altered at the posttranslational level. Several agents have been used in an attempt to inhibit fibrosis. Glucocorticosteroids can inhibit the synthesis of collagen and other proteins. Penicillamin and beta-aminopropionitrile can inhibit the cross-linking of collagen, the latter via and inhibition of lysyl oxidase. Prolin analogues as cis-hydroxyprolin have been used to inhibit the intracellular post-translational enzymes. Colchicine disrupts microtubules and slows the rate of secretion of procollagen and it may also increase the secretion of collagenase.
There is currently much interest in phytochemicals as bioactive components of food. The roles of fruit, vegetables and red wine in disease prevention have been attributed, in part, to the antioxidant properties of their constituent polyphenols (vitamins E and C, and the carotenoids). Recent studies have shown that many dietary polyphenolic constituents derived from plants are more effective antioxidants in vitro than vitamins E or C, and thus might contribute significantly to the protective effects in vivo. It is now possible to establish the antioxidant activities of plant-derived flavonoids in the aqueous and lipophilic phases, and to assess the extent to which the total antioxidant potentials of wine and tea can be accounted for by the activities of individual polyphenols.
Long-term exposure to UV radiation leads to skin ageing and may initiate carcinogenesis. In both cases immunosuppressive activity of UV radiation plays an important role. The aim of the study is to present polyphenols commonly seen in flora and their properties protecting the skin from the damaging influence of UV rays. Polyphenols are a group of compounds which are present in plants. Their common features are: the ring structure of a molecule, hydroxyl groups in the rings and a conjugated double bond system. Such structure makes polyphenols active antioxidants. They also demonstrate anti-immunosuppressive properties.
Publisher Summary This chapter discusses the analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Analyses of the Folin-Ciocalteu (FC) type are convenient, simple, and require only common equipment and have produced a large body of comparable data. Under proper conditions, the assay is inclusive of monophenols and gives predictable reactions with the types of phenols found in nature. Because different phenols react to different degrees, expression of the results as a single number—such as milligrams per liter gallic acid equivalence—is necessarily arbitrary. Because the reaction is independent, quantitative, and predictable, analysis of a mixture of phenols can be recalculated in terms of any other standard. The assay measures all compounds readily oxidizable under the reaction conditions and its very inclusiveness allows certain substances to also react that are either not phenols or seldom thought of as phenols (e.g., proteins). Judicious use of the assay—with consideration of potential interferences in particular samples and prior study if necessary—can lead to very informative results. Aggregate analysis of this type is an important supplement to and often more informative than reems of data difficult to summarize from various techniques, such as high-performance liquid chromatography (HPLC) that separate a large number of individual compounds .The predictable reaction of components in a mixture makes it possible to determine a single reactant by other means and to calculate its contribution to the total FC phenol content. Relative insensitivity of the FC analysis to many adsorbents and precipitants makes differential assay—before and after several different treatments—informative.
Chlorogenic acids are polyphenolic compounds that occur ubiquitously in foods of plant origin. They are quinic acid esters of hydroxycinnamic acid.Recently, naturally occurring plant phenolics have attracted considerable attention in relation to their physiological potential. Depending upon the conditions, phenolic compounds can be either beneficial or detrimental to biological processes.We comprehensively summarized chlorogenic acids and related compounds in absorption, metabolism and biological activity. Chlorogenic, caffeic and quinic acids are well absorbed in humans and rats. Metabolic transformations of chlorogenic acids in the human system may be crucial for their biological effect.The antioxidant activities of chlorogenic acids are preserved by inhibiting the formation of reactive oxygen species or by scavenging them. As a result, chlorogenic acidsmay play beneficial role in the prevention of certain oxidative diseases. The observed pharmacological activities of medicinal plants relate to the chlorogenic acidsconstituents of them. Despite the abundance of biological data demonstrating the antioxidant activities of chlorogenic acids, it remains controversial whether these compounds are potent antioxidants or pro-oxidants. Chlorogenic and caffeic acids switch from anti- to pro-oxidant activity, depending on their concentration, on the presence of free transition metal ions, or on their redox status.