Determination of in vitro antioxidant and UV-protecting activity of aqueous and ethanolic extracts from Galinsoga parviflora and Galinsoga quadriradiata herb

doi:10.1016/j.jphotobiol.2015.06.010

Journal of Photochemistry and Photobiology B: Biology

Volume 149, August 2015, Pages 189-195

https://doi.org/10.1016/j.jphotobiol.2015.06.010 Get rights and content

Highlights

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Aqueous extracts from the Galinsoga herb are effective as photoprotectors.
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The activity may be related to the content of caffeoylglucaric acids.
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Ethanolic extracts from the herb have cytotoxic activity.

Abstract

Galinsoga species are used in folk medicine as anti-inflammatory agents and accelerators for wound healing. They also have reported antioxidant activity. We examined aqueous and ethanolic extracts derived from the Galinsoga herb as potential photoprotectors, as the role of reactive oxygen species (ROS) has implicated in skin damage. The extracts used in the study were standardized by determining the sum of flavonoids, and the amount of caffeic acid and its derivatives. The antioxidant activity of the extracts was evaluated by examining the scavenging of two radicals (O₂⁻ and H₂O₂) generated in cell-free systems. We also examined the effect on ROS generation by human skin fibroblasts after UV irradiation. In addition we determined the cytotoxicity of the extracts and their protective effect against damage caused by UV irradiation (MTT test, LDH release test and staining with annexine V-FITC/PI).

Our findings show that the ethanolic extracts from the herb have cytotoxic effects, while the aqueous extracts from Galinsoga herb have protective activity, in part due to their ability to inhibit ROS generation.

In the conclusion the aqueous extracts from the both tested species may be effective as photoprotectors.

Introduction

Sunlight is composed of wavelengths from ultraviolet light (UVR) through infrared to visible light. The most harmful to the skin is ultraviolet light, which is divided into three categories dependent on wavelengths: UVA (315–400 nm), UVB (280–315 nm) and UVC (100–280 nm). While UVC is absorbed by the ozone layer and scarcely reaches the Earth, UVA and UVB rays, in addition to their positive effects, can also act negatively on living organisms. UVA penetrates deep into the epidermidis and dermis of the skin. UVB acts mainly in the epidermal basal cell layer of the skin, but is more genotoxic and about 1000 times more capable of causing sunburn than UVA. UVA and UVB rays provoke free radical production and can induce a significant decrease in skin antioxidants, leading to the skin being less able to protect itself against free radicals generated after sunlight exposure. Amongst others mostly singlet oxygen – an excited state of oxygen – is generated, which is a very powerful oxidant with a relatively long lifetime. However, in some reactions a superoxide anion is also produced, followed by dismutation to hydrogen peroxide. Sunlight mainly penetrates different layers of the skin, so they are therefore the most vulnerable to the adverse effects of excessive sun exposure. Chronic exposure to UVR leads to photoaging, immunosuppression, photocarcinogenesis, and the exacerbation of photodermatoses [1], [2], [3].

Galinsoga ciliata Raf. Blake (Galinsoga quadriradiata Ruiz et Pav) like G. parviflora Cav., are annual herbs belonging to the Asteraceae family, which originate from the Andes region. The chemical composition, activity and use of both species tends to be similar. Flavonoids: patulitrin (patuletin-7-O-β-d-glucoside), quercimeritrin (quercetin-7-O-β-d-glucoside), quercetagetrin (quercetagetin-7-O-β-d-glucoside), luteolin 7-β-d-glucopyranoside, apigenin 7-β-d-glucoside, galinsoside A (5′-hydroxy-7-methoxyflavanone 2′-O-β-d-glucopyranoside), galinsoside B (3′,4′-dihydroxy-7-methoxyflavanone 5-O-β-d-glucopyranoside), 7,3′,4′-trihydroxyflavanone and 3,5,7,3′,4′-pentahydroxyflavanone, and phenolic acids and depsides: vanilic, isovanilic, p-coumaric, p-hydroxybensoic, o-hydroxyphenyloacetic, caffeic, chlorogenic acid and caffeoylglucaric acids, among other compounds have been found in extracts of Galinsoga herb [4], [5], [6], [7], [8], [9], by phytochemical analysis.

Galinsoga species extracts are applied topically to skin, to treat dermatological diseases, such as eczemas, lichens and hard-healing-wounds, and also to treat snakebites. Orally they are administered to treat flu and common colds [10], [11], [12], [13], [14].

Antioxidants, also of natural origin, and polyphenolic compounds derived from plants, are reported among the factors that protect the skin from the harmful effects of solar radiation [15], [16], [17], [18], [19], [20], [21]. The Galinsoga herb is traditionally used in various skin diseases, it contains polyphenols, and has antioxidant activity which has been shown in previous studies [22], [23]. The aim of our present study was to determine the antioxidant activity of aqueous and ethanolic extracts (most commonly used in medicine) derived from the Galinsoga herb against two main ROS, O₂⁻ and H₂O₂, produced by UV radiation. The extracts used in the study were standardized by determining the content of flavonoids, and the sum of caffeic acid derivatives (caffeoyl glucarates and caffeoly quinic acids) and caffeic acid using HPLC method. The antioxidant activity of these extracts was evaluated by examining the capacity of scavenging of radicals generated in cell-free systems. We also investigated the effect on ROS generation by human skin fibroblasts after UV irradiation. In addition we also tested for the potential cytotoxicity of the extracts and their protective effect against damage caused by UV irradiation.

Section snippets

Chemicals

Dichlorofluorescin diacetate (DCFH₂-DA), formic acid 99%, horseradish peroxidase (HRP), hydrogen peroxide (H₂O₂), luminol, nitrobluetetrazolium (NBT), quercetin, xanthine, and xanthine oxidase were purchased from Sigma–Aldrich Chemie GmbH (Steinheim, Germany). Chlorogenic acid was purchased from Fluka Chemica (Buchs, Switzerland). Triton X-100 solution and Cytotoxicity Detection Kit (LDH) were purchased from Roche Diagnostics (Mannheim, Germany). Annexin V Apoptosis Detection Kit was purchased

Results and discussion

Initially we standardized the extracts by determining the content of flavonoids, and the sum of caffeic acid derivatives (caffeoyl glucarates and caffeoly quinic acids) and caffeic acid using HPLC method. This analysis revealed that the highest content of flavonoids was determined in G. parviflora ethanolic extract (685.4 ± 69.6 mg Q g⁻¹), while the highest content of sum of caffeic acid derivatives and caffeic acid in G. parviflora aqueous extract (44.3 ± 2.7 mg ChA g⁻¹) (The results of quantitative

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Galinsoga parviflora (Cav.) is a member of the Asteraceae family traditionally used for treatment of various ailments such as malaria, flu, cold, colorectal cancer, liver problems and inflammation. The medicinal properties of G. parviflora are due to the presence of various secondary metabolites including flavonoids, saponins, terpenoids and tannins. The literature survey revealed that G. parviflora possesses several pharmacological properties such as antibacterial, antifungal, antioxidant and antidiabetic. This review systematically discusses the potential of G. parviflora for managing medical conditions. The information is collected from various online databases such as Google Scholar, ScienceDirect, Springer, Web of Science, Plant of the World Online and PubMed. Among other information provided in this review, ethnomedicinal uses, phytochemistry and pharmacological activities are discussed extensively. Additonally, the potential benefits, challenges and future opportunities are presented.
Defense potential of secondary metabolites in medicinal plants under UV-B stress
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Ultraviolet-B (UV-B) radiation has, for many decades now, been widely studied with respect to its consequences on plant and animal health. Though according to NASA, the ozone hole is on its way to recovery, it will still be a considerable time before UV-B levels reach pre-industrial limits. Thus, for the present, excessive UV-B reaching the Earth is a cause for concern, and UV-B related human ailments are on the rise. Plants produce various secondary metabolites as one of the defense strategies under UV-B. They provide photoprotection via their UV-B screening effects and by quenching the reactive oxygen- and nitrogen species produced under UV-B influence. These properties of plant secondary metabolites (PSMs) are being increasingly recognized and made use of in sunscreens and cosmetics, and pharma- and nutraceuticals are gradually becoming a part of the regular diet. Secondary metabolites derived from medicinal plants (alkaloids, terpenoids, and phenolics) are a source of pharmaceuticals, nutraceuticals, as well as more rigorously tested and regulated drugs. These metabolites have been implicated in providing protection not only to plants under the influence of UV-B, but also to animals/animal cell lines, when the innate defenses in the latter are not adequate under UV-B-induced damage. The present review focuses on the defense potential of secondary metabolites derived from medicinal plants in both plants and animals. In plants, the concentrations of the alkaloids, terpenes/terpenoids, and phenolics have been discussed under UV-B irradiation as well as the fate of the genes and enzymes involved in their biosynthetic pathways. Their role in providing protection to animal models subjected to UV-B has been subsequently elucidated. Finally, we discuss the possible futuristic scenarios and implications for plant, animal, and human health pertaining to the defense potential of these secondary metabolites under UV-B radiation-mediated damages.
Caffeic acid derivatives isolated from Galinsoga parviflora herb protected human dermal fibroblasts from UVA-radiation
2019, Phytomedicine
Among solar radiation, ultraviolet light is the most harmful for the skin, because of intracellular reactive oxygen species formation, leading to oxidative stress, cell damage and apoptosis. Crucial role in skin protection against oxidative stress play antioxidant enzymes regulated by Nrf2 transcription factor. Some plant-derived polyphenols are known to protect skin fibroblasts against UV through induction of Nrf2-dependent antioxidant genes expression.
We previously found out that water extracts from Galinsoga sp. herb protected human dermal fibroblasts against UVA-induced oxidative stress and apoptosis. However, which compounds were responsible for such protective action remained unclear. Here, we investigated photoprotective potential and mechanism of action of two main isolated compounds, 2,3,5(2,4,5)-tricaffeoylaltraric acid and 2,4(3,5)-dicaffeoylglucaric acid, on human dermal fibroblasts (NHDF).
NHDF cells were pretreated with tested compounds (6.25–50 µM) and irradiated with UVA (25 J/cm²). Intracellular ROS and GSH level, cell viability, cell membrane integrity and apoptosis were measured. HO-1 protein expression and Nrf2 transcription factor activation were also assessed.
Cells pretreated with tested compounds prior to UVA showed inhibition of intracellular ROS formation and increase of GSH level. Significant increase of cell viability was also observed, as well as decrease of LDH release and a the rate of apoptotic cells in comparison to untreated cells. Furthermore, tested compounds increased HO-1 expression and activated the Nrf2 transcription factor in NHDF cells.
Present study demonstrated that caffeic acid derivatives present in Galinsoga parviflora herb, in particular tricaffeoylaltraric acid may protect dermal fibroblasts against UVA-induced oxidative stress through activation of intracellular antioxidative system. Such caffeic acid derivatives are bioactive compounds which might prevent UV-induced photoageing and photocarcinogenesis.
Analysis of chlorogenic acids isomers and caffeic acid in 89 herbal infusions (tea)
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In results reported in the literature, Figueroa et al. (2014) showed values between 67.0 and 104 μg∙100 mL−1 in mint infusions. Only a few studies have reported the presence of caffeic acid in gallant soldier, and currently there are no studies that quantify this compound in an infusion of this plant (Bazylko et al., 2015a, 2015b). From the analyzed infusions, 71% showed 3-CQ values between 1.7 μg∙100 mL−1 for juniper (Juniperus communis) and 14000 μg∙100 mL−1for yerba mate (Ilex paraguariensis), and five showed content above 1000 μg∙100 mL−1.
Several plants used to prepare aqueous beverages (teas) are known for their health benefits due to the presence of bioactive compounds, such as isomers of 3-caffeoylquinic (3-CQ), 4-caffeoylquinic (4-CQ), 5-caffeoylquinic (5-CQ), 3,4-dicaffeoylquinic (3,4-DQ), 3,5-dicaffeoylquinic (3,5-DQ), 4,5-dicaffeoylquinic (4,5-DQ) and caffeic acid. This study analyses the presence of these compounds in infusions from 89 plants. We acquired these plants from public markets and analyzed them using ultra-performance liquid chromatography. We found the listed chlorogenic acids in 93% of infusions. The infusions containing highest concentrations were yerba mate (Ilex paraguariensis), with 52.6 mg in 100 mL of infusion, followed by assa-peixe (Boehmeria caudata), white tea (Camellia sinensis), winter’s bark (Drimys winteri), green tea (Camellia sinensis), and elderflower (Sambucus nigra), with 6.0 mg in 100 mL of infusion. Several plant infusions had not been previously studied. Compared to other foods and beverages, these infusions are considered potential sources of chlorogenic acids.
Herbal extracts, lichens and biomolecules as natural photo-protection alternatives to synthetic UV filters. A systematic review
2016, Fitoterapia
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Taken together, these results suggested D. sessile D. Don as a potential ingredient in order to prevent skin photoaging and treat UVB-induced skin damage. As reported in another study [69], in vitro antioxidant and UV-protecting studies were performed on aqueous and ethanolic extracts from Galinsoga parviflora and Galinsoga quadriradiata. While ethanolic extracts showed cytotoxic effects and improved ROS generation after UVB irradiation, both aqueous extracts (for both plants) exhibited interesting photoprotective activities.
Besides the unquestionable positive effects of solar exposure for human health, UV rays have been widely investigated for toxicology aspects related to excessive UVB and UVA doses, which involve sunburns, skin aging, DNA skin damage and tumorigenesis. At present, synthetic and mineral sunscreens are used to protect against these damages but several natural molecules can provide UV protection, including also synergic effect or enhanced photo stability. Although a large number of herbal extracts and plant origin molecules can deserve potential applications, most of the study reported utilizes different method and different strategies of investigation, making thus difficult to understand the real versus claimed potential. This is possibly one of the reasons why, beside the large body of literature there are no officially approved natural commercial sun-filter but a consistent number of commercially available solar products (sunscreen) on the market that contain herbal derivatives. In this review we have evaluated the papers appeared in the last 15 years and we have critically collected the most significant data. Several databases, namely Scifinder, Pubmed, Google Scholar, ISI-Web of Science and Scopus, were used as literature sources; excluding patents and symposium or congress papers. Only articles in the English language have been selected. New formulation, new skin delivery systems, skin penetration enhancers and boosters are most likely the next frontier of investigation in order to better understand the role of whole herbal extracts in exerting their photo protective activity.
Caffeic acid derivatives isolated from the aerial parts of Galinsoga parviflora and their effect on inhibiting oxidative burst in human neutrophils
2016, Phytochemistry Letters
Citation Excerpt :
As 2,3,5- or 2,4,5-tricaffeoylaltraric acid (15) appears to be one of the most active compounds (inhibition of ROS production: IC50 = 0.7 μM), and additionally is one of the dominating compounds in gallant soldier extract, it may be treated as a marker of raw material/extract quality. The result obtained also explains which compounds are responsible for the observed antioxidant activities of G. parviflorum extracts (Bazylko et al., 2015b; Bazylko et al., 2012). We have also performed a study concerning the potential anti-inflammatory activity of 15, by determination of interleukin 8 (IL-8) and tumor necrosis factor (TNF-α) release from stimulated neutrophil.
Four new caffeoyl -glucaric and -altraric acid derivatives along with eleven known compounds were isolated from aerial parts of Galinsonga parviflora. Their structures were elucidated by high-resolution spectroscopic studies. The four new compounds were determined as being 2,3,4,5-tetracaffeoylglucaric acid (1), 2,4,5-tricaffeoylglucaric acid (2), 2,3,4- or 3,4,5-tricaffeoylaltraric acid (3) and 2,3(4,5)-dicaffeoylaltraric acid (4). A reliable criterion for the determination of the linkage position of caffeic acids moieties in glucaric acid derivatives has been proposed, on the basis of detailed analysis of the respective J-couplings, including substitution and solvent influence on the observed values. All hexaric acids derivatives appeared as inhibitors of reactive oxygen species production by stimulated neutrophils.

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Determination of in vitro antioxidant and UV-protecting activity of aqueous and ethanolic extracts from Galinsoga parviflora and Galinsoga quadriradiata herb

Highlights

Abstract

Introduction

Section snippets

Chemicals

Results and discussion

J. Autoimmun.

Bioresour. Technol.

Phytochem. Lett.

J. Ethnopharmacol.

J. Ethnopharmacol.

Arch. Biochem. Biophys.

Regul. Toxicol. Pharm.

J. Photochem. Photobiol. B

Plant Sci.

Biochem. Pharmacol.

Phytomedicine

Phytochemistry

J. Ethnopharmacol.

Oxidants and human disease: some new concepts

FASEB J.

Ultraviolet light induced alteration to the skin

Biomed. Pap. Med. Fac. Univ. Palacky Olomouc Czech Repub.