Hypericum alpestre extract exhibits in vitro and in vivo anticancer properties by regulating the cellular antioxidant system and metabolic pathway of L-arginine.

Conventional treatment methods are not effective enough to fight the rapid increase in cancer cases. The interest is increasing in the investigation of herbal sources for the development of new anticancer therapeutics. This study aims to investigate the antitumor capacity of Hypericum alpestre (H. alpestre) extract in vitro and in vivo, either alone or in combination with the inhibitors of the  l-arginine/polyamine/nitric oxide (NO) pathway, and to characterize its active phytochemicals using advanced chromatographic techniques. Our previous reports suggest beneficial effects of the arginase inhibitor NG-hydroxy-nor- l-arginine and NO inhibitor NG-nitro-Larginine methyl ester in the treatment of breast cancer via downregulation of polyamine and NO synthesis. Here, the antitumor properties of H. alpestre and its combinations were explored in vivo, in a rat model of mammary gland carcinogenesis induced by subcutaneous injection of 7,12-dimethylbenz[a]anthracene. The study revealed strong antiradical activity of H. alpestre aerial part extract in chemical (DPPH/ABTS) tests. In the in vitro antioxidant activity test, the H. alpestre extract demonstrated pro-oxidant characteristics in human colorectal (HT29) cells, which were contingent upon the hemostatic condition of the cells. The H. alpestre extract expressed a cytotoxic effect on HT29 and breast cancer (MCF-7) cells measured by the MTT test. According to comet assay results, H. alpestre extract did not exhibit genotoxic activity nor possessed antigenotoxic properties in HT29 cells. Overall, 233 substances have been identified and annotated in H. alpestre extract using the LC-Q-Orbitrap HRMS system. In vivo experiments using rat breast cancer models revealed that the H. alpestre extract activated the antioxidant enzymes in the liver, brain, and tumors. H. alpestre combined with chemotherapeutic agents attenuated cancer-like histological alterations and showed significant reductions in tumor blood vessel area. Thus, either alone or in combination with Nω -OH-nor- l-arginine and Nω -nitro- l-arginine methyl ester, H. alpestre extract exhibits pro- and antioxidant, antiangiogenic, and cytotoxic effects.

Comparative Analysis of Phytochemical Profiles and Selected Biological Activities of Various Morphological Parts of Ligustrum vulgare.

Ligustrum vulgare (LV), widely cultivated in Europe and often used in hedges, has been historically recognized in folk medicine for its potential health benefits. This study focused on exploring the untargeted identification of secondary metabolites in ethanol extracts (70% v/v) from different morphological parts (young shoots, leaves, flowers and fruits) of LV at various stages of plant development, using ultra-high-performance liquid chromatography with high-resolution mass spectrometry (UHPLC-HRMS). Additionally, the selected biological activities (antioxidant activity, cyclooxygenase-2 inhibition (COX-2), α-amylase inhibition and cytotoxicity) of the tested extracts were determined. Untargeted metabolomics showed that LV extracts were a rich source of phenylethanoid compounds, flavonoids, iridoids and their derivatives. The flowers of LV had the highest content of oleuropein (33.43 ± 2.48 mg/g d.w.). The lowest antioxidant activity was obtained for ripe and post-seasonal fruits, while in the case of other samples, the activity was at a similar level. All tested extracts showed α-amylase and COX-2 inhibitory activity. In addition, LV extracts showed strong antiproliferative properties in colorectal (HT29) and liver (HepG2) cancer cell lines. The obtained results show the difference in the content of bioactive compounds in various morphological parts of Ligustrum vulgare. These differences may influence the multifaceted medicinal potential of this plant.

The Action Mechanisms, Anti-Cancer and Antibiotic-Modulation Potential of Vaccinium myrtillus L. Extract.

BACKGROUND: Herbal medicinal products containing Vaccinium myrtillus L. (bilberry) fruits and fruit extracts are widely available in the market. Although bilberry leaves and stems are considered as bio-waste, they contain much higher levels of phenolic compounds than fruits. The study aimed to investigate the antimicrobial and anticancer potential of aerial part extracts from Vaccinium myrtillus L. (V. myrtillus, VM) plants harvested at high altitudes in Armenian landscape and characterize the bioactive phytochemicals. MATERIAL AND METHODS: For evaluation of antioxidant properties, chemical-based tests (total phenolic and flavonoid content, and antiradical activity in 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) tests) and cellular antioxidant activity (CAA) assay were applied. Genotoxicity and anticancer properties of the extract alone and in combination with fluorouracil were explored in human cancer and normal cell lines. Antibacterial properties of V. myrtillus extract alone and in combination with antibiotics, as well as their effect on proton-flux rate through cell membrane were explored on bacterial strains. The characterization of active phytochemicals was done using Liquid Chromatography-Quadrupole-Orbitrap High-Resolution Mass Spectrometry (LC-Q-Orbitrap HRMS). RESULTS: The V. myrtillus aerial part extract demonstrated promising antioxidant properties in all tests. The selective cytotoxic activity was documented against various cancer cell lines (human colon adenocarcinoma (HT29), human breast cancer (MCF-7) and human cervical carcinoma (HeLa)), while it did not inhibit the growth of tested human normal primary renal mixed epithelial cells (HREC) even at 10-fold higher concentrations. The extract did not have genotoxic properties in comet assay making it a potential source for the development of anticancer preparations. The investigated extract did not directly inhibit the growth of Escherichia coli (E. coli) and Salmonella typhimurium (S. typhimurium) strains at up to 1 mg/mL concentration. However, V. myrtillus extract enhanced the kanamycin intake and increased its efficiency against E. coli strain. The phytochemical characterization of the extract showed the presence of different groups of phenolics. CONCLUSIONS: Based on obtained data, we suggest the aerial parts of the V. myrtillus plant as an alternative source of bioactive natural products for food supplements, nutraceuticals, functional foods and medicine.

Natural Products Counteracting Cardiotoxicity during Cancer Chemotherapy: The Special Case of Doxorubicin, a Comprehensive Review.

Cardiotoxicity is a frequent undesirable phenomenon observed during oncological treatment that limits the therapeutic dose of antitumor drugs and thus may decrease the effectiveness of cancer eradication. Almost all antitumor drugs exhibit toxic properties towards cardiac muscle. One of the underlying causes of cardiotoxicity is the stimulation of oxidative stress by chemotherapy. This suggests that an appropriately designed diet or dietary supplements based on edible plants rich in antioxidants could decrease the toxicity of antitumor drugs and diminish the risk of cardiac failure. This comprehensive review compares the cardioprotective efficacy of edible plant extracts and foodborne phytochemicals whose beneficial activity was demonstrated in various models in vivo and in vitro. The studies selected for this review concentrated on a therapy frequently applied in cancer, anthracycline antibiotic-doxorubicin-as the oxidative stress- and cardiotoxicity-inducing agent.

Comparative evaluation of different methods for determining phytochemicals and antioxidant activity in products containing betalains - Verification of beetroot samples.

This study presents methods that can be used to assess the health quality of products containing betalains. The paper compares and verifies data on the phytochemical composition of three different pigmented beetroot cultivars using spectrophotometric, HPLC-DAD, HPTLC and LC-Q-Orbitrap-HRMS techniques. Additionally, we compared the total antioxidant activity in both the cell-free and cellular systems. Betalain contribution to antioxidant activity was also determined using post-column derivatization and it was found that in the case of red beetroot it is about 50%. Photometric measurements are recommended for a simple and inexpensive analysis of the total betacyanin and betaxanthin content. Liquid chromatography techniques produced more precise information on phytochemical composition in the tested samples. The combination of liquid chromatography with high-resolution mass spectrometry produced the largest amount of quantitative and qualitative data; in beetroot samples sixty-four phytochemicals have been identified therefore, this approach is recommended for more detailed metabolomics studies.

The Relationship between Phytochemical Composition and Biological Activities of Differently Pigmented Varieties of Berry Fruits; Comparison between Embedded in Food Matrix and Isolated Anthocyanins.

The aim of this research was to correlate the composition of phenolic compounds and bioactivities (antioxidant, cytotoxic, antigenotoxic, and influence on selected enzymatic activities) exhibited by extracts from differently pigmented raspberry (yellow and red), grape (white and red), mulberry (white and black), and currant (white, red, and black) varieties. It was presumed that phytocomplexes of the same species will be similar while differing significantly only by the content of anthocyanins in colored varieties, which will impact biological properties. Additionally, to compare food matrix embedded and isolated anthocyanins, the influence of purified solution, in concentrations matching the total concentration of anthocyanins in appropriate colored plant samples, on investigated biological activities was studied. The phenolic compound composition and antioxidant profiles showed that anthocyanin content was correlated only with antioxidant capacity of tested plant extracts. The other determined biological activities failed to reveal any simple relationship between chemopreventive potential and anthocyanin composition in fruits studied nor any similarity to biological properties of isolated cyanidin-3-O-glucoside. These observations indirectly support the food synergy concept, that points to interactions between different phytochemicals as a factor deciding about the final bioactivity of edible plants.

The Caucasian flora: a still-to-be-discovered rich source of antioxidants.

Cellular redox homeostasis is a state of balance between the formation of Usually Reactive Oxygen and / or Nitrogen Species (ROS/RNS), endogenous antioxidant defence systems, and exogenous dietary antioxidants. The disturbance of redox homeostasis, by the overproduction of endogenous ROS/RNS, may increase the risk of development of so-called civilisation diseases. The solution seems to be either the increased production of endogenous or consumption of exogenous antioxidants. Plant-borne antioxidants act via different chemical and molecular mechanisms, such as decreasing the level of oxidative damage in cells directly by reacting with ROS/RNS or indirectly - by inhibition of the activity and expression of free radical generating enzymes or by enhancing the activity or expression of intracellular antioxidant defence enzymes. Despite the fact that the Caucasian flora is rich of health promoting edible/medicinal plants, recent studies concerning the biological activity of these plants are very scarce. This review is summarising the state-of-art on the health-promoting potential of plants representing the Caucasian flora, whose antioxidant capacity have been investigated in various in vitro models.

Prophylaxis of Non-communicable Diseases: Why Fruits and Vegetables may be Better Chemopreventive Agents than Dietary Supplements Based on Isolated Phytochemicals?

The World Health Organization (WHO) report from 2014 documented that non-communicable socalled civilization diseases such as cardiovascular disease, chronic respiratory diseases, cancer or type 2 diabetes are responsible for over 50% of all premature deaths in the world. Research carried out over the past 20 years has provided data suggesting that diet is an essential factor influencing the risk of development of these diseases. The increasing knowledge on chemopreventive properties of certain food ingredients, in particular, those of plant origin, opened the discussion on the possibility to use edible plants or their active components in the prevention of these chronic diseases. Health-promoting properties of plant foods are associated with the presence of secondary metabolites that can affect many biological mechanisms of critical importance to the proper functioning of the human organism. Particularly, there have been numerous investigations indicating strong physiological effects of bioactive plant phenols belonging to the flavonoid family. These observations initiated mass production of dietary supplements containing flavonoids commercialized under the name antioxidants, even if their chemical properties did not justify such a term. However, epidemiological studies revealed that isolated bioactive phytochemicals are not as effective as fruits and vegetables containing these substances whereas they are of interest of the functional food industry. In this paper, the critical assessment of reasons for this turn of events has been attempted and the concept of food synergy has been suggested as a future strategy of dietary chemoprevention.

Phytochemical composition and biological activities of differently pigmented cabbage (Brassica oleracea var. capitata) and cauliflower (Brassica oleracea var. botrytis) varieties.

BACKGROUND: Brassica plants contain a wide spectrum of bioactive components that are responsible for their health-promoting potential such as vitamins, polyphenols and glucosinolates. This study attempted to relate the composition of bioactive phytochemicals and chosen biological activities (antioxidant, cytotoxic, anti-genotoxic, and influence on enzymatic activities) for extracts from differently pigmented cabbage (white and red) and cauliflower (white and purple) varieties. The assumption was that tested varieties of the same plant would exhibit similar chemical composition differing mostly in anthocyanin content and that the latter will be reflected in biological activity. RESULTS: Profiles of antioxidants obtained using post-column derivatization with ABTS radical confirmed, that the content and composition of anthocyanins is strongly correlated with the antioxidant capacity of tested plant extracts measured by spectrophotometric methods (ABTS, FC, DPPH, FRAP), and Cellular Antioxidant Activity (CAA) test. The results of determinations of other biological activities showed that opposite to purified bioactive phytochemicals, in the case of actual plant foods, there was no simple relationship between anthocyanin content and chemopreventive potential. CONCLUSION: Obtained results suggest that there must be some kind of interaction between different phytochemicals, which decides on the final health promoting activity of edible plants as suggested by for example the food synergy concept. © 2019 Society of Chemical Industry.

The comparison of betalain composition and chosen biological activities for differently pigmented prickly pear (Opuntia ficus-indica) and beetroot (Beta vulgaris) varieties.

Betalains are a group of plant originated pigments with chemopreventive potential. The aim of this study was to relate the composition of betalains and chosen biological activities (antioxidant, cytotoxic, anti-genotoxic and influence on enzymatic activities) for extracts from differently pigmented varieties of prickly pear (yellow, orange and red) and beetroot (white and red). The assumption was that phytocomplexes of tested varieties of the same plant species would exhibit generally similar chemical composition differing mostly in betalain content, which will be reflected by their biological activity. Betalain composition analysis and antioxidant profiles confirmed that the content and composition of these pigments is strongly correlated with the antioxidant activity of tested plant extracts measured by spectrophotometric methods and CAA test. However, the results of determinations of other biological activities showed that in the case of actual plant foods, there was no simple relationship between betalain content or composition and chemopreventive potential.

Determination of antioxidantactivity of phytochemicals in cellular models by fluorescence/luminescence methods.

As soon as the role of Reactive Oxygen Species (ROS) in so-called civilization diseases, which include non-infectious chronic diseases such as cancer, diabetes or high blood pressure has been discovered, and the possibility of employing antioxidants as a remedy for these diseases have been proposed, scientists developed a broad spectrum of methods to determine antioxidant activity of pure chemicals and plant extracts, as well as dietary supplements. Most of these methods are based on simple redox reactions between antioxidant and ROS (for example ABTS, DPPH, or FRAP tests). However, chemical methods of assessing antioxidant activity are rarely biologically relevant. They do not mirror the real effect of antioxidants in living organisms, because they are used in non-physiological conditions of temperature and pH; neither they take metabolism nor intracellular transport under consideration. The perfect model for assessment of antioxidant activity in living organisms would be human or animal model, but such determinations are very complicated and often ambiguous. The current best alternative to chemical and human tests are assays employing cell culture models being less expensive than human tests, yet still reflecting biological systems more convincingly than chemical assays. Cellular antioxidant assays are performed under physiological pH and temperature, but most importantly, they take metabolism and intracellular transport under consideration. In this review, we present cellular tests used to determine antioxidant activity that are based on luminescence and fluorescence methods.