Protective effect of bilberry (Vaccinium myrtillus L.) on cisplatin induced ovarian damage in rat

Cisplatin is one of the most effective chemotherapeutic agents but injury may occur at higher doses. The aim of this study was to investigate the effect of bilberry on cisplatin induced toxic effects in rat ovary. Twenty-one female Wistar–Albino rats were utilized to form three groups: In group 1 (control group), each rat received intraperitoneal injection of 1 mL of 0.9 % NaCl saline solution during 10-days. In group 2 (cisplatin group), a single dose of 7.5 mg/kg b.w. cisplatin was given. In group 3 (cisplatin + bilberry group), a single dose of 7.5 mg/kg cisplatin and bilberry at 200 mg/kg b.w. were given for 10 days. Ovaries were surgically removed in all groups and prepared for biochemical and light microscopic investigations at the examination times. Malondialdehyde (MDA) levels and activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) of tissue samples were measured. Histopathological damages in cisplatin administrated rats were seen such as severe edema, vascular congestion, hemorrhage and follicular degeneration in the ovary tissue. Moderate pathological alterations were observed in rats treated with bilberry plus cisplatin. Cisplatin administration significantly increased MDA production and decreased SOD, CAT, GPx and GST activities in the ovarian tissue when compared to the control group (p < 0.05). Cisplatin + bilberry administration increased antioxidant enzymes activities and reduced MDA levels. Bilberry administration seems to reduce the cisplatin induced ovarian toxicity thus it alleviates free radical damage. But it dose not protect completely rat ovary tissues.     

Isolation of high quality RNA from bilberry (Vaccinium myrtillus L.) fruit

A simple and efficient method is described for isolating high quality RNA from bilberry fruit. The procedure is based on the use of hexadecyltrimethyl ammonium bromide (CTAB), polyvinylpyrrolidone (PVP), and β-mercaptoethanol in an extraction buffer in order to eliminate the polysaccharides and prevent the oxidation of phenolic compounds. This method is a modification of the one described for pine trees, and yields high-quality RNA suitable for cDNA based methodologies. This method is applicable for a variety of plant tissues.     

Ascorbic acid metabolism during bilberry (Vaccinium myrtillus L.) fruit development

Bilberry (Vaccinium myrtillus L.) possesses a high antioxidant capacity in berries due to the presence of anthocyanins and ascorbic acid (AsA). Accumulation of AsA and the expression of the genes encoding the enzymes of the main AsA biosynthetic route and of the ascorbate-glutathione cycle, as well as the activities of the enzymes involved in AsA oxidation and recycling were investigated for the first time during the development and ripening of bilberry fruit. The results showed that the AsA level remained relatively stable during fruit maturation. The expression of the genes encoding the key enzymes in the AsA main biosynthetic route showed consistent trends with each other as well as with AsA levels, especially during the first stages of fruit ripening. The expression of genes and activities of the enzyme involved in the AsA oxidation and recycling route showed more prominent developmental stage-dependent changes during the ripening process. Different patterns of activity were found among the studied enzymes and the results were, for some enzymes, in accordance with AsA levels. In fully ripe berries, both AsA content and gene expression were significantly higher in skin than in pulp.     

Responses to harvesting intensity in a clonal dwarf shrub,the bilberry (Vaccinium myrtillus L.)

The effects of herbivory were simulated on stands of bilberry (Vaccinium myrtillus L.) in a boreal Empetrum-Myrtillus type forest. Five harvesting intensities were used (0% (A), 25% (B), 50% (C), 100% (D) of the bilberry ramets or all the ramets of all species (E)). Density and biomass of the stands, and growth of the ramets were monitored for five growth seasons, from 1986 to 1990. After damage new ramets emerged rapidly from dormant buds at the base of removed ramets. Between 70 and 97% of the density relative to the control level was regained by the final harvest. However, only between 11 and 64% of the biomass relative to the control level was recovered. Clipping reduced the branch growth, both in the new ramets and in the new parts of the old, unclipped ramets. Severe treatments (D and E) decreased the growth more than did light harvesting (B and C). Survival and fecundity of the ramets were not affected. The result therefore suggested that the bilberry is not able to recover totally from severe herbivory damage between the years of peak rodent population that shows a 3–4 year cycle. Nevertheless, extreme grazing pressure is rare in boreal ecosystems as alternative food is available for herbivores, and predators also limit the herbivore population. Hence the bilberry exhibits moderate tolerance of the usual level of herbivory damage.     

Extracts of bilberry (Vaccinium myrtillus L.) fruits improve liver steatosis and injury in mice by preventing lipid accumulation and cell death

ABSTRACT

Bilberry has been reported to have anti-oxidant and anti-inflammatory properties. We studied the effect of bilberry (Vaccinium myrtillus L.) fruits extracts (BEs) on the pathogenesis caused by lipid accumulation in fatty liver and non-alcoholic steatohepatitis (NASH). 5 μg/ml of BEs was enough to suppress lipid accumulation in the fatty liver model of the mouse hepatic AML12 cells. BEs increased cell viability and anti-oxidant capacity, presumably by activating (phosphorylating) Akt/STAT3 and inducing MnSOD/catalase. BEs also significantly reduced Rubicon and induced p62/SQSTM1, possibly contributing to reduce cellular lipids (lipophagy). When the mice were fed supplemented with BEs (5% or 10%, w/w), hepatic steatosis, injury, and hypercholesterolemia/hyperglycemia were significantly improved. Furthermore, histological and cytokine studies indicated that BEs possibly suppress hepatic inflammation (hepatitis) and fibrosis. Therefore, BEs improved liver steatosis and injury, and potentially suppress fibrosis by suppressing inflammatory response, which therefore may prevent the progression of fatty liver to NASH.     

Recovery of the bilberry (Vaccinium Myrtillus L.) from artificial spring and summer frost

Vegetative and sexual recovery of bilberry (Vaccinium myrtillus L.) was monitored in the field for three growing seasons after artificially applied spring and early summer frost. Bilberry recovered vegetatively (density and biomass) by vigorous production of new ramets and by production of large shoots in the damaged ramets. Recovery did not occur sexually (production of flowers), however. Summer frost was slightly more detrimental than the spring frost, but the difference was significant only in the percentage of flowering ramets. The results showed that bilberry is buffered against moderate frost injury. The rapid vegetative recovery guarantees survival of the plants after frost damage, whereas sexual reproduction may need accumulation of resources for many seasons. Consequently, the ability of bilberry to recover from sporadic frosts increases its potentiality to persist in a changing environment, where increasing temperature leads to earlier snowmelt in the spring, and increases the risk of frost injuries in plants that overwinter under snow.     

Predicting biomass of bilberry (Vaccinium myrtillus) using rank distribution and root-to-shoot ratio models

Plant Ecology - Investigation of relationships between biomass of different plant compartments is important because their varying turnover rates affect the amount and composition of aboveground and...     

Chemopreventive effect of bilberry (Vaccinium myrtillus) against cisplatin-induced oxidative stress and DNA damage as shown by the comet assay in peripheral blood of rats

The aim of this study was to evaluate the chemopreventive effect of bilberry on cisplatin induced oxidative stress and DNA damage in rat blood. Twenty-one female Wistar-Albino rats were divided into three groups: group I — untreated; group II — treated with cisplatin (single dose 7.5 mg/kg b.w.); and group III — treated with cisplatin (single dose 7.5 mg/kg b.w.) and bilberry (200 mg/kg b.w. for 10 days). Antioxidant enzyme systems including superoxide dismutase, catalase, glutathione peroxidase and the level of malondialdehyde (MDA) that might occur on erythrocytes have been determined and single cell gel electrophoresis (comet) was utilized to evaluate the DNA damage in lymphocytes. Treatment with cisplatin has increased the levels of MDA and decreased antioxidant enzymes in erythrocytes. Comet assay showed significantly higher values at dose of 7.5 mg/kg cisplatin as the result of oxidative DNA damage when compared to the control group. Cisplatin treatment with bilberry resulted in a highly significant (P < 0.05) decreased in the lymphocytes DNA when compared to the cisplatin group. Bilberry has been effective on antioxidant enzyme systems and MDA level and significantly decreased the comets. Our results indicate that bilberry is capable of preventing genotoxic and cytotoxic damage caused by cisplatin in peripheral blood cells in rats.     

Severe salt stress in Vaccinium myrtillus (L.) in response to Na+ ion toxicity

NaCl is spread on the northern roads as a winter de-icing agent. Na⁺ stress may occur in roadside forests especially during growing season, because snowmelt increases Na⁺ concentrations in the roadside forest soil. To simulate summertime Na⁺ stress and its effect on viability and anthocyanin concentrations, bilberry (Vaccinium myrtillus L.) plants were subjected to NaCl (0, 6, 30 and 60 g m⁻²) in a boreal mesic heath forest in northern Finland (65°N). It was hypothesised that Na⁺ stress decreases tissue water content (TWC) in below- and aboveground stems, where resulting water stress increases anthocyanin level for osmotic regulation. Uptake of Na⁺ from the soil to the below- and aboveground stems was detected by a fast sequential atomic absorption spectrometry. Na⁺ accumulated in belowground stems, but it was translocated into the aboveground stems to a lesser extent. At the end of the growing season, TWC increased in belowground stems and decreased in aboveground stems. Also anthocyanin concentrations decreased in aboveground stems. The viability of the aboveground stems decreased to 30% along with the Na⁺ accumulation. Despite the strong viability response in aboveground stems, TWC was constant in the middle of the growing season and decreased relatively little at the end of the growing season. It is thus proposed that direct effect of Na⁺ ions, rather than Na⁺-induced water stress, is the primary reason behind the strong viability response and a severe salt stress in bilberry.     

Substitution of Vaccinium myrtillus L. for Aronia melanocarpa (Michx.) Elliott in a commercial batch

Abstract

The fruits of Vaccinium myrtillus and Aronia melanocarpa are utilized in pharmaceutical and food preparations as rich sources of anthocyanoside pigments. However, these two drugs should not be considered equivalent because their chemical profiles are different. In this work, we report a case of substitution of V. myrtillus for A. melanocarpa in a commercial batch. The comparative morphological investigation of the two drugs is described.     

Protective effect of an acidic glycoprotein obtained from culture of Chlorella vulgaris against myelosuppression by 5-fluorouracil

 An acidic glycoprotein prepared from a culture of Chlorella vulgaris (CVS) was examined for its protective effect on 5-fluorouracil(5FU)-induced myelosuppression and indigenous infection in mice. Subcutaneous administration of CVS greatly reduced the mortality of non-tumor-bearing mice given a high dose of 5FU, and could increase the LD₅₀ value of 5FU for these mice. After 5FU treatment, indigenous infection developed probably as a result of the impairment of the host defense system. CVS reduced the incidence of indigenous infections and this effect was attributable to the acceleration of recovery from 5FU-induced myelosuppression. Early recovery of hematopoietic stem cells, or cells responding to interleukin-3 or granulocyte/macrophage-colony-stimulating factor, was especially observed in the bone marrow of CVS-treated mice on days 4 – 9 after the injection of 5FU. When tumor-bearing mice were given CVS during treatment with 5FU, CVS prolonged the survival of mice without affecting the antitumor activity of 5FU. In addition, CVS was itself shown to exert an antitumor effect. These results suggested that CVS may be beneficial for the alleviation of side-effects in cancer chemotherapy without affecting the antitumor activity of the chemotherapeutic agent. Received: 15 August 1995 / Accepted: 23 April 1996     

Anthocyanins and glutathione S-transferase activities in response to low temperature and frost hardening in Vaccinium myrtillus (L.)

Anthocyanin (Acy) contents and GST activities of bilberry (Vaccinium myrtillus L.) were investigated in two experiments conducted in June (Exp. I: active growth) and in August September (Exp. II: beginning of frost hardening) in Northern Finland (65 degrees N). Bilberry plants were subjected to +2 degrees C and +18 degrees C in Exp. I or +5/0 degrees C (day/night) and +18/+13 degrees C (day/night) in Exp. II. GST activities were assessed using either 1-chloro-2,4-dinitrobenzene (CDNB) or trans-cinnamic acid (tCA) as substrates. We found temperature to have no effect on Acy during either active growth or frost hardening. Acy increased several-fold from active growth to the beginning of frost hardening, but no increment was observed during the development of frost hardening. This suggests a role of Acy in photoprotection at low temperatures rather than their direct involvement in the development of freezing tolerance. The lack of response of GST activity to frost hardening and to temperature in autumn may indicate an indirect role of GSTs in frost hardening as protective enzymes. GST activity was the same with the two substrates studied (CDNB, tCA), supporting the assumption that GSTs could catalyze reactions with endogenous phenylpropanoids.     

Comparison of two methods used to analyse lipid peroxidation from Vaccinium myrtillus (L.) during snow removal, reacclimation and cold acclimation.

Malondialdehyde (MDA) concentration is a widely used method to analyse lipid peroxidation in biological material. In plant tissues, however, certain compounds (anthocyanins, carbohydrates) may interfere with measurements which may lead to an overestimation of the MDA levels. Two methods were compared for analysing lipid peroxidation, either uncorrected or corrected for interfering compounds. The comparison was performed in three separate experiments with respect to cold treatments (snow removal in winter, reacclimation in summer and cold acclimation in autumn) in bilberry (Vaccinium myrtillus L.). During winter and autumn the methods seem to measure different compounds, but during active growth in the summer the difference between the methods was less. This is obviously due to carbohydrates which act as cryoprotectants and increase in concentration during cold acclimation as well as due to the anthocyanins. It is thus suggested that the validity of the uncorrected method to measure MDA and thereby lipid peroxidation is best in plant tissue which is in an active growth state.     

Experimental evidence from Cantabrian mountain heathlands suggests new recommendations for management of Vaccinium myrtillus L

Background: Management actions in mountain heathlands oriented to increasing the cover of Vaccinium myrtillus promote mowing of Calluna vulgaris to avoid competition. However, such action ignores the fact that plant–plant interactions range from competition to facilitation under different stress conditions.

Aims: To test whether the interactions between these two species are related to competition or facilitation, which would change the perspective for their management in mountain heathlands.

Methods: A total of 40 experimental plots located in a montane heathland (northern Spain) were selected. Calluna or Vaccinium plants were either cut at ground level or not cut, and plots were fertilized with nitrogen or not in a three-factorial design. Vegetative and reproductive shoot mass and length of the current year’s shoots were estimated in two consecutive years for both species. Herbivory on Vaccinium by browsing ungulates was compared in control plots and plots where Calluna had been cut 2 yrs ago.

Results: The two study species co-occurred in 72.4% of the sampled plots, although Calluna was more abundant. Vaccinium growth was greater in plots with Calluna than in plots where Calluna had been cut. Browsing on Vaccinium was also higher in plots were Calluna had been cut, and the effect of fertilization was only significant on the reproductive biomass of Calluna.

Conclusions: Contrary to initial proposals, our results suggest that Vaccinium does not always benefit from the cutting of Calluna in heathlands. Thus, the management of Cantabrian mountain heathlands should consider maintaining Calluna in order to facilitate the growth of Vaccinium.     

Protective effect of soybean against hepatocarcinogenesis induced by DL-ethionine

There has been increasing interest in the value of using soybean to delay or reduce the tumor incidence. This study was undertaken to investigate the possible protective effects of soybean against hepatocarcinogenesis induced by DL-ethionine. Accordingly, we measured biochemical changes occurring in serum and liver of rats treated with DL-ethionine in the presence or absence of soybean. Male albino rats were fed a control diet containing the hepatocarcinogen, DL-ethionine, or the control diet plus soybean 30%, or the control diet plus soybean plus DL-ethionine 0.25% for three months and then returned to a control diet for up to nine months. Rats fed a control diet plus DL-ethionine showed a gradual decrease in liver DNA, RNA, total protein, and liver weight and enzyme activities of liver transaminases (GOT and GPT) and alkaline phosphatase over the 7-month study period. This was followed by a large increase in the liver parameters at the end of the 9(th) month, except for 5'-nucleotidase and glucose-6-phosphatase that showed a large decrease. On the other hand, a gradual increase in the serum enzyme activities of GOT, GPT, 5-nucleotidase, alkaline phosphatase, and in the albumin/globulin (A/G) ratio is observed in the group of rats fed a control diet plus DL-ethionine compared to the control group over 8 months, and this was followed by a large increase in all serum parameters studied at nine-months. The administration of 30% soybean to the rat diet in addition to DL-ethionine maintained all parameters studied at near control values until the end of the 9(th) month. This study suggests that soybean has a protective effect against the hepatocarcinogenesis induced by DL-ethionine.     

Protective effect of jasmonic acid and potassium against cadmium stress in peas (Pisum sativum L.)

A combination of mineral nutrients and plant growth regulators should be assessed to improve crop performance under various abiotic stresses. There is a need to include plant growth regulators in fertilization regime of various crops along with essential mineral nutrients, especially when they are irrigated with polluted water with higher levels of heavy metals. The performance of pea was evaluated under cadmium (Cd) stress coupled with potassium (K) and jasmonic acid (JA) supplementation. The Cd stress (50 μM) was applied to soil (sandy loam) grown pea plants as basal dose after a month of sowing. The control and stressed plants were then supplemented with K (5 M), JA (0.5 mM) and their collective application along with control as distilled water. Cd stress showed a marked reduction in growth pattern, however, the collective supplementation sufficiently improved the growth pattern of stressed peas plants as evidenced by improvement in shoot length (cm), root length (cm), number of leaves per plant, leaf area (cm²), plant fresh and dry weight (gm). Potassium application under Cd stress significantly enhanced internodal distance (cm) while the number of seeds per pod and relative water contents remained nonsignificant. The applied treatment (JA + K) under Cd stress prominently improved enzymatic activities, which were measured as nitrate reductase activity (NRA), nitrite reductase activity (NiRA), superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT). Cd stress impacted the biochemical profile by enhancing antioxidant capacity (AC), antioxidant activity (AA), total phenols (TP), while reducing total soluble protein (TSP), chlorophyll ‘a’, chlorophyll ‘b’ and carotenoids. The combined application of JA and K under Cd stress enhanced AC, AA, TP, Chl a and b, TSP and carotenoids. The results indicate that foliar application of JA and K efficiently negated the harmful effects of Cd stress on peas.     

Activation of flavonoid biosynthesis by solar radiation in bilberry (<Emphasis Type="Italic">Vaccinium myrtillus</Emphasis> L.) leaves

The effect of solar radiation on flavonoid biosynthesis was studied in bilberry (Vaccinium myrtillus L.) leaves. Expression of flavonoid pathway genes of bilberry was studied in the upper leaves of bilberry, exposed to direct sunlight, in the shaded leaves growing lower in the same plants and in fruits. Bilberry-specific digoxigenin–dUTP-labeled cDNA fragments of five genes from the general phenylpropanoid pathway coding phenylalanine ammonia-lyase and from the flavonoid pathway coding chalcone synthase, flavanone 3-hydroxylase, dihydroflavonol 4-reductase, and anthocyanidin synthase were used as probes in gene expression analysis. Anthocyanins, catechins, proanthocyanidins, flavonols and hydroxycinnamic acids from the leaves and fruits were identified and quantified using high-performance liquid chromatography combined with a diode array detector. An increase in the expression of the studied flavonoid pathway genes was observed in leaves growing under direct sun exposure. Also, the concentrations of anthocyanins, catechins, flavonols and hydroxycinnamic acids were higher in the leaves exposed to direct sunlight. However, the concentration of polymeric procyanidins was lower in sun-exposed leaves, whereas that of prodelphinidins was slightly increased. The results give further support for the protective role of flavonoids and hydroxy cinnamic acids against high solar radiation in plants. Also, the roles of different flavonoid compounds as a defense against stress caused by sun exposure is discussed.Abbreviations ANS Anthocyanidin synthase - CHS Chalcone synthase - DFR Dihydroflavonol 4-reductase - F3H Flavanone 3-hydroxylase - GPD Glyceraldehyde-3-phosphate dehydrogenase - PAL Phenylalanine ammonia-lyase     

Protective effect of propolis against oxidative stress and immunosuppression induced by oxytetracycline in rainbow trout (Oncorhynchus mykiss, W.)

The aim of this study was to investigate the effects of propolis on oxytetracycline (OTC)-induced oxidative stress and immunosuppression in fish. OTC (100 mg per kg?1 body weight) was orally administered to fish for 14 days. A significant elevation in the level of malondialdehyde, as an index of lipid peroxidation, and reductions in antioxidant enzyme activities (superoxide dismutase, catalase, and glutathione peroxidase) and low molecular weight antioxidant (reduced glutathione) levels were observed in the blood, liver, kidney, spleen, and heart tissues of OTC-treated fish. OTC also had a suppressive effect on specific and non-specific immune system parameters, such as leucocyte counts, oxidative radical production (nitrobluetetrazolium activity), total plasma protein and immunoglobulin levels, and phagocytic activity. Pre-treatment, post-treatment, and simultaneous treatment with propolis (50 mg per kg?1 body weight, orally) attenuated the OTC-induced oxidative stress by significantly decreasing the levels of malondialdehyde in tissues. In addition, propolis significantly increased the level of reduced glutathione and the catalase, glutathione peroxidase, and superoxide dismutase activities. Upon the administration of propolis, the suppressed immune system parameters were significantly increased in fish treated with OTC. The present results suggest that pre-treatment, post-treatment, and simultaneous administration of propolis might alleviate OTC-induced oxidative stress and immunosuppression.     

Protective effect of acetyl-L-carnitine against oxidative stress induced by antiretroviral drugs

Both HIV infection per se and antiretroviral drugs might contribute to oxidative stress and mitochondrial dysfunctions. In this study we assess zidovudine, stavudine and didanosine on U937 and CEM cell lines. All these drugs induced apoptosis and increased intracellular hydrogen peroxide but not superoxide anions. The addition of acetyl-l-carnitine (ALC) was able to prevent the pro-oxidant effect of the drugs tested. Supplementation with ALC, deficient in certain cohorts of HIV-infected individuals, especially on high active antiretroviral therapy regimen, has been associated with favourable effects. These data suggest that one of these effects could be a direct anti-oxidant action.