Fig 3 - uploaded by Pompei Bolfa
Content may be subject to copyright.
(A) Epidermal hyperplasia, intracellular and intercellular oedema and induction of sunburn cells at 24 hours after UVB exposure in four mice experimental groups (arrows indicate representative sunburn cells). Cv and BM extracts were administered topically (4 mg polyphenols/cm 2 ), 30 min before of UVB irradiation. Animals were sacrificed at 24 hours after UVB exposure; skin biopsies were sampled, preserved in 10% buffered formalin and processed for histopatological investigations. Representative photomicrographs of Trichrome-Masson and haematoxylin-eosin stain from each treatment group, n=10, with an original magnification x 400 (Scale bar=50 µm). (B) A total of ten microscopic fields were counted. The results were expressed as the mean percent of positive cells per field ±S.D.
Source publication
Solar ultraviolet radiation (UV) is a major cause of non-melanoma skin cancer in humans. Photochemoprevention with natural products represents a simple but very effective strategy in the management of cutaneous neoplasia. The study investigated the protective activity of Calluna vulgaris (Cv) and red grape seeds (Vitis vinifera L, Burgund Mare vari...
Contexts in source publication
Context 1
... mouse skin was examined for the presence of sunburn cells 24 hours after irradiation (240 mJ/cm 2 ). Representative photomicrographs of haematoxylin-eosin and Trichrome-Masson stained sections obtained from unirradiated, UV-irradiated and treated skin are shown in Fig. 3. Examination revealed the presence of sunburn cells, which exhibited the classic patterns of apoptosis: pyknotic nuclei and condensed cytoplasm and intracellular oedema (Fig. 3). Haematoxylin and eosin-stained sections of unirradiated mouse skin were used as controls and showed only few cells undergoing normal cell death, with a random ...
Context 2
... 2 ). Representative photomicrographs of haematoxylin-eosin and Trichrome-Masson stained sections obtained from unirradiated, UV-irradiated and treated skin are shown in Fig. 3. Examination revealed the presence of sunburn cells, which exhibited the classic patterns of apoptosis: pyknotic nuclei and condensed cytoplasm and intracellular oedema (Fig. 3). Haematoxylin and eosin-stained sections of unirradiated mouse skin were used as controls and showed only few cells undergoing normal cell death, with a random distribution (1.50±0.57%). In the UV- irradiated skin, the number of sunburn cells peaked at 24 hours after irradiation (25.50±5.44%; p<0.0002). Some of the sunburn cells ...
Context 3
... Pretreatment with Cv extract insignificantly reduced the number of sunburn cells (19.20±12.64%; 25% inhibition; p>0.05), while BM extract administration determined an important reduction of apoptotic cells number (10.38±2.92%; 60% inhibition; p<0.02). Keratinocytes vacuolization and hydropic changes, characteristic for UVB irradiated mouse skin (Fig. 3, panel A) were less severe in the skin samples pretreated with Cv and BM extracts. In the dermis, the inflammatory reaction was scanty perivascular (Fig. ...
Context 4
... an important reduction of apoptotic cells number (10.38±2.92%; 60% inhibition; p<0.02). Keratinocytes vacuolization and hydropic changes, characteristic for UVB irradiated mouse skin (Fig. 3, panel A) were less severe in the skin samples pretreated with Cv and BM extracts. In the dermis, the inflammatory reaction was scanty perivascular (Fig. ...
Citations
... BM demonstrated antioxidant properties (directly correlated with its content in polyphenols) and photoprotective effects on UVB-induced reactive oxygen species production (54). ...
Melanoma, a cancer derived from melanocytes is very difficult to treat, especially in advanced cases. There are several encouraging studies of the efficacy of photodynamic therapy (PDT) in melanoma. However, PDT has to overcome the main defense mechanisms like: defects in the apoptotic pathways, pigmentation, sequestration of the photosensitisers (PS) inside melanosomes and increased oxidative stress defense.
Two meso-substituted porphyrins, meso-5,10,15,20-tetrakis (4-hydroxyphenyl) porphyrin (THOPP) and meso-5-(4-hydroxyphenyl)-10, 15, 20-tris (4-methoxyphenyl) porphyrin (THOMPP) were used as PS to investigate several mechanisms underlining the PDT anti-melanoma effects, on a lightly pigmented melanoma cell line (WM35), in vitro. γH2AX foci formation (a measure of DNA double strand brakes) was used for the assessment of DNA damage by means of immune-fluorescence and western blot. Cytoskeleton alterations were detected by phalloidin staining. Tyrosinase activity and melanin pigment were quantified by spectrophotometry, tyrosinase protein by western blot, total peroxidase activity by resorurfin reaction (Amplex Red).
PDT induced high levels of DNA damage, cytoskeleton alterations and enhanced pigmentogenesis. THOPP mediated PDT was the most efficient. The melanogenesis stimulated by PDT was directly correlated to the PDT induced cellular damage and provided no protection against therapy. Thus, PDT induced melanogenesis combined with severe DNA damage was able to overcome the mechanisms of resistance and increased the efficiency of PDT in WM35 melanoma cells. These results are encouraging for a possible use of PDT, as an adjuvant therapy in lightly pigmented melanomas.
... In parallel to the above study, a pretreatment combination of hydroethanolic extracts of BM with Calluna vulgaris (4 mg/40 μL/cm 2 ) had protective effects against a single dose of UV-B irradiation (240 mJ/cm 2 ) and decreased IL-6 and TNF-α levels compared to UV-B group in the same mouse species. It seems that the grape extract works like a sunscreen and could modulate inflammatory and apoptotic responses to UV-B irradiation in mice (Filip et al., 2011a). ...
Vitis vinifera fruit (grape) contains various phenolic compounds, flavonoids and stilbenes. In recent years, active constituents found in the fruits, seeds, stems, skin and pomaces of grapes have been identified and some have been studied. In this review, we summarize the active constituents of different parts of V. vinifera and their pharmacological effects including skin protection, antioxidant, antibacterial, anticancer, antiinflammatory and antidiabetic activities, as well as hepatoprotective, cardioprotective and neuroprotective effects in experimental studies published after our 2009 review. Clinical and toxicity studies have also been examined. Copyright © 2016 John Wiley & Sons, Ltd.
... BM demonstrated antioxidant properties (directly correlated with its content in polyphenols) and photoprotective effects on UVB-induced reactive oxygen species production (54). ...
We studied the effect of grape seed extract Burgund Mare (BM) on oral carcinogenesis and compared it with that of curcumin (CU). Wistar rats were divided into six groups (n = 10): 4-nitro-quinoline-1-oxide (4NQO) oral carcinogenesis was induced to groups 1 - 5; groups 2 and 3 received BM and CU respectively during initiation and groups 4 and 5 BM and CU during post-initiation of carcinogenesis; group 6 represented the negative control group. Total malondialdehyde (MDA) and reduced glutathione (GSH) were assayed fluorometrically in oral tissue (gingival, jugal, palatal, lingual mucosa) and serum. Histopathological exam was performed and a dysplasia score given to each oral mucosal lesion. Ki67, cyclin D1, p63, Bcl2 and p53 were immunohistochemically evaluated. BM and CU reduced tissue MDA values elevated by 4NQO (P = 0.000). The difference between CU and BM effect was significant in the initiation (P = 0.02) but not in the post-initiation phase of carcinogenesis (P = 0.58). Tissue GSH levels decreased by 4NQO (P < 0.001) were not significantly modified by BM or CU. Serum MDA levels increased by 4NQO (P = 0.000) were significantly lowered by CU (P = 0.04) and BM (P = 0.04) during initiation and by CU during post-initiation of carcinogenesis (P = 0.01). CU was more potent than BM during post-initiation of carcinogenesis (P = 0.01). Serum GSH lowered by 4NQO (P = 0.55) was significantly decreased by BM and CU (P < 0.012), with no significant difference between groups receiving BM or CU. Moderate dysplasia was the most advanced dysplasia induced and gingival localization the most frequent. Both BM and CU lowered dysplasia scores, with BM being the most efficient during post-initiation of carcinogenesis (P = 0.001). Ki67, cyclin D1, p63, Bcl2 and p53 expression increased with dysplasia scores. BM showed chemopreventive properties during initiation and post-initiation of oral carcinogenesis, reducing local and general oxidative stress and the intensity of dysplasia. During post-initiation of carcinogenesis BM and CU exhibited similar effects.
... In a previous report, we showed the protective effect of Cv extract against UVB radiation in vitro, on a HaCaT cell line, by reducing oxidative stress and DNA damage and increasing cell survival [8]. Also, in vivo, topical application of Cv extract on mouse skin markedly decreased UVB-induced oxidative stress, apoptosis and UVB-induced cytokines levels (IL-6, TNF-a) [9]. Pretreatment with Cv extract reduced NF-jB and MMP-9 activity after single UVB exposure, but after multiple doses of UVB, in 10 consecutive days, Cv activated NF-jB and increased MMP-9 activity [10]. ...
... However, Cv had the opposite effect, e.g. activated NF-jB and increased MMP-9 activity after multiple, consecutive UVB exposures [9]. ...
... ). Sunburn cells were identified in the epidermis as cells with a homogeneous, densely-staining glossy eosinophilic cytoplasm and a small hyperchromatic condensed pyknotic nucleus that can readily be seen with routine HE-stained histological sections of the skin using light microscopy (Lu et al., 2005). The mean percentage ± SD of sunburn cells per field in the epidermis (basal plus suprabasal layer) was calculated from the number of these cells divided to the total number of cells, by counting 10 HPF, as previously reported (Filip et al., 2011a,b). ...
... itro. The use of grape antioxidants are promising against a broad range of cancer cells by targeting epidermal growth factor receptor (EGFR) and its downstream pathways, inhibiting over-expression of COX-2 and prostaglandin E2 receptors, or modifying oestrogen receptor pathways, resulting in cell cycle arrest and apoptosis (Zhou and Raffoul, 2012). Filip et al. (2011) in their studies on Photoprotective effects of two natural products on ultraviolet B-induced oxidative stress and apoptosis in SKH-1 mouse skin reported that their results suggest that Calluna vulgaris and Vitis vinifera extracts might be chemopreventive candidates for reducing UV-induced risk for skin cancer. Currants and Sultanas (Vit ...
Medicinal plants have been used in healthcare since time immemorial. Studies have been carried out globally to verify their efficacy and some of the findings have led to the production of plant-based medicines. The global market value of medicinal plant products exceeds $100 billion per annum. This paper discusses the role, contributions and usefulness of medicinal plants in tackling the diseases of public health importance, with particular emphasis on the current strategic approaches to disease prevention. A comparison is drawn between the 'whole population' and 'high-risk' strategies. The usefulness of the common-factor approach as a method of engaging other health promoters in propagating the ideals of medicinal plants is highlighted. The place of medicinal plants in preventing common diseases is further examined under the five core principles of the Primary Health Care (PHC) approach. Medicinal plants play vital roles in disease prevention and their promotion and use fit into all existing prevention strategies. However, conscious efforts need to be made to properly identify, recognise and position medicinal plants in the design and implementation of these strategies. These approaches present interesting and emerging perspectives in the field of medicinal plants. Recommendations are proposed for strategising the future role and place for medicinal plants in disease prevention.
... In our previous works, we found that crude grape seed extract, Burgund Mare variety (BM), protected SKH-1 hairless mice skin against UVB-induced damage by reducing the lipid peroxidation and nitric oxide production, and also by diminishing caspase-3 activity (Filip et al., 2011b) and cytokine levels (IL-6 and TNF-a) (Filip et al., 2011a). In addition, BM extract inhibited formation of sunburn cells and cyclobutane pyrimidine dimers (CPDs) induced by single or multiple doses of UVB irradiation (Filip et al., 2011c). ...
... The antioxidant capacity of BM was higher than in gallic acid, measured both by electron paramagnetic resonance technique (k BM = 0.017, k gallic acid = 0.013) (Filip et al., 2011c) or by using ABTS (52.89 ± 0.02 vs. 6.3 ± 0.4 eq. mM Trolox) (Filip et al., 2011a) and free radical DPPH tests (0.072 ± 0.002 vs. 0.083 ± 0.004 mmol/mmol DPPH) (Dicu et al., 2010). The previously published HPLC data (Perde-Schrepler et al., 2013) indicated the presence of procyanidin B, catechin hydrate, epigallocatechin, epicatechin and gallic acid as main compounds. ...
... The BM extract was applied topically on the skin in a dose of 4 mg TP/cm 2 , in vehicle (acetone 40%), 30 min before each UVB exposure. UVB irradiation was performed with Waldmann UV 181 broadband UVB source as described before (Filip et al., 2011a,b). The UVB emission was monitored with a radiometer Variocontrol radiometer (Waldmann GmbH, Germany). ...
... Further, both in vitro (6) and in vivo studies (7-9) revealed antioxidant and anti-inflammatory effects of Cv extract. Recently, we demonstrated that pretreatment with Cv extract reduced lipid peroxides and nitric oxide generation and inhibited the UVBinduced apoptosis and inflammation in mice (8,9) as well as the formation of DNA photolesions in HaCaT keratinocytes (10). These effects were assigned to the high content of polyphenols identified in the extract composition (10). ...
... 2,2-diphenyl-l-picryl-hydrazyl (DPPH)·free radical assay (19) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate (ABTS) test were used to evaluate the antioxidant activity (8). ...
... The extract was applied topically on the skin in a dose of 2.5 (low dose) respectively 4 mg/cm 2 (high dose), in vehicle, 30 min before each UVB exposure. UVB irradiation was performed with Waldmann UV 181 broadband UV-B source as described previously (8,9). The UVB emission was monitored with a radiometer Variocontrol radiometer (Waldmann GmbH, Germany). ...
Photochemoprevention with natural products represents a new concept in the attempt to reduce the occurrence of skin cancer. However, the molecular mechanisms caused by ultraviolet light exposure remain still unclear. The aim of the study was to assess the mechanisms involved in the action of a Calluna vulgaris (Cv) extract, administered in single or multiple doses (10 consecutive days), on UVB-induced skin damage in SKH-1 hairless mice. The extract was topically applied 30 min before each UVB exposure in two doses (2.5 and 4 mg total polyphenolic content/40 μl/cm(2)). At 24 hours after the last treatment, total mitogen-activated protein kinase (p44/42MAPkinase, ERK 1/2), nuclear factor-κB (phospho-NF-κB p65), matrix metalloproteinases (MMP-2, MMP-9) and metalloproteinase inhibitor 1 (TIMP-1) levels were measured in skin using enzyme-linked immunosorbent assay (ELISA). MMP-2 and -9 activities were additionally evaluated by zymography. One topical application of Cv extract reduced the secretion (p<0.004) and inhibited MMP-9 activity UVB-mediated (54% inhibition) via inhibition of NF-κB activation (68% inhibition). In multiple UVB exposures, both doses of Cv extract induced the increase of ERK 1/2 level in correlation with activation of NF-κB and reduced the secretion (p<0.04) and activation of MMP-9 (62% inhibition). Pretreatment with Cv diminished the MMP-2 protein secretion only in one dose UVB-irradiated group (p<0.0001) and decreased TIMP-1 level (p<0.001). These results demonstrated the dual behavior of Cv extract in skin protection against single versus multiple doses of UVB irradiation.
... Similarly, resveratrol could induce phosphorylation of CAT, POD and Cu/Zn-SOD and by this way enhanced their activities without modifying their expression nor abundance. At the opposite, resveratrol was shown to reduce endothelial oxidative stress by modulating the gene expression of SOD1 and GPx1 (63), which is unlikely to occur in our present case because of the enucleated status of erythrocytes. ...
Doxorubicin (Dox), a widely used antitumor anthracycline antibiotic, plays an undisputed key role in the treatment of many neoplasic diseases. In this study, the protective role of resveratrol against Dox-induced erythrocytes and plasma toxicity has been evaluated in rats. Animals were treated with resveratrol (25 mg/kg b.w.) by intraperitoneal injection during 8 days. At the 4(th) day of treatment, rats were intraperitoneally injected with a single dose of Dox (20 mg/kg b.w.). At the end of the treatment, blood samples were collected following standard procedure and processed for oxidative stress parameters (malondialdehyde (MDA), carbonyl protein, free iron, calcium and H(2)O(2) levels), transaminases and antioxidant enzymes as catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Data showed that Dox drastically increased erythrocytes and plasma MDA, free iron, H(2)O(2) and carbonyl proteins but decreased calcium levels and also decreased erythrocytes CAT, POD and SOD activity. Besides, Dox decreased plasma CAT and SOD but unexpectedly increased POD activity. Dox also increased plasma ALT and AST levels and decreased them into erythrocytes. Co-treatment with resveratrol counteracted almost all Dox's effects. In conclusion, Dox induced a pro-oxidative stress into erythrocytes and resveratrol exerted real antioxidant properties which can be attributed, at least in part, to free iron and calcium modulation.
... 32 Other data from animal studies support the photoprotective nature of grape-seed extract. 33,34 These are extremely safe and common compounds that are unlikely to have sustainability issues or cause adverse effects. Many other herbal extracts, many of them probably redox modulators, have been reported to counteract photoaging in vitro (Table 1). ...
Some herbal agents taken orally appear to be able to reduce local and systemic negative effects of excessive ultraviolet (UV) light exposure. What is notable in this category is the well-researched Phlebodium aureum (golden serpent fern), frequently referred to in the literature by an old name Polypodium leucatomos. Sufficiently high oral doses of Camellia sinensis (green tea), as well as topical applications, also appear to be photoprotective. In addition, Panax ginseng (Asian ginseng) may be helpful as a systemic immunomodulator to prevent the harm done by excessive UV light on local and general immune function. What is much less well-established is the potential of the immunomodulating fungus Cordyceps sinensis to have this same benefit. Redox modulators such as propolis and proanthocyanidins (from Pinus maritima or Vitis vinifera) have also shown the ability topically to prevent sunburn and UV-related toxicities. More research is needed but natural products look promising as photoprotectives. Note: The next article related to this topic will discuss herbal photosensitizers, or agents that enhance the therapeutic properties of UV light.
