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1.
Molecules ; 22(4)2017 Apr 12.
Article in English | MEDLINE | ID: mdl-28417940

ABSTRACT

Uncaria tomentosa is a woody vine with a long history of use in traditional Peruvian medicine and nowadays supplements containing this vine as ingredient are available. Immunomodulating, anti-inflammatory and anticancer properties of Uncaria tomentosa have been suggested and attributed mainly to the presence of tetracyclic or pentacyclic oxindole alkaloids. However, the synergic action of different compounds occurring in extracts and modulation of redox processes may significantly influence the anticancer activity of Uncaria tomentosa. The aim of the present study was to investigate for the first time the cytotoxic effects of the tetracyclic alkaloids free aqueous extract (decoction) of dried Uncaria tomentosa leaf blades in normal and cancer cells, and to assess the effect of the tested extract on cisplatin (CDDP) cytotoxicity. Tested Uncaria tomentosa extract was not cytotoxic for NHDF cells, but demonstrated cytotoxic effect against HepG2 cells. The extract increased ROS production in HepG2 cells, which resulted in decreased GSH level, leading to apoptosis of these cells through activation of caspase-3 and caspase-7. A reduction of NF-κB active form was observed in cancer cells. In normal cells the extract did not affect ROS production, GSH level and NF-κB activity, and maintained cell viability. HepG2 cells incubation with Uncaria tomentosa decoction and simultaneously with CDDP resulted in an increase in CDPP cytotoxic activity against HepG2, while under the same conditions Uncaria tomentosa prevents NHDF cell viability reduction due to CDDP. The results indicate that Uncaria tomentosa leaves decoction modulates differently cancer and normal cells oxidative metabolism and, enhanced cytotoxicity of CDDP against cancer cells and at the same time increased normal healthy cells resistance to cisplatin. Further studies are needed to confirm our observations and to describe underlying molecular mechanism, and the potential usefulness of Uncaria tomentosa decoction in adjuvant therapy for cancer.


Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Cat's Claw/chemistry , Oxidation-Reduction/drug effects , Plant Extracts/pharmacology , Plant Leaves/chemistry , Reactive Oxygen Species/metabolism , Antineoplastic Agents, Phytogenic/chemistry , Apoptosis/drug effects , Caspase 3/metabolism , Caspase 7/metabolism , Cell Line , Cell Survival/drug effects , Cisplatin/pharmacology , Hep G2 Cells , Humans , NF-kappa B/metabolism , Plant Extracts/chemistry
2.
J Sci Food Agric ; 94(13): 2718-25, 2014 Oct.
Article in English | MEDLINE | ID: mdl-24532295

ABSTRACT

BACKGROUND: In this study we examined the influence of various forms of selenium (organic and inorganic) on the vivacity of Hericium erinaceum mycelium and structural changes and ultrastructure occurring during its development in submerged culture. RESULTS: The mycelium was grown on sodium selenite (Na2SeO3), Selol (with 20 and 50 g kg⁻¹ Se, respectively) and a mixture of Na2SeO3 and Selol. Samples of the mycelium were collected on day 3 and day 24 of the incubation and viewed under an electron microscope. Selol at concentration 20 g kg⁻¹ did not cause any damage to the cell ultrastructure, but it contributed to the thickening of the cell wall, which implied an influence on polysaccharide production. In the other cases, degradation changes appeared in the protoplasm and the thickness of the cell wall did not increase. CONCLUSION: The nature of the effect exerted by various sources of selenium in the culture medium on the formation of polysaccharides probably results from the differences in their chemical composition and differences in the toxicity of these compounds towards the cells, but is also connected with the decomposition of the wall surrounding degraded fungal cells.


Subject(s)
Basidiomycota/metabolism , Food Quality , Mycelium/metabolism , Oxidative Stress , Selenium Compounds/metabolism , Sodium Selenite/metabolism , Basidiomycota/growth & development , Basidiomycota/ultrastructure , Cell Wall/metabolism , Cell Wall/ultrastructure , Cytoplasm/metabolism , Cytoplasm/ultrastructure , Dietary Supplements/adverse effects , Food, Fortified/adverse effects , Fungal Polysaccharides/biosynthesis , Hydrolysis , Microbial Viability , Microscopy, Electron, Transmission , Mycelium/growth & development , Mycelium/ultrastructure , Poland , Selenium Compounds/adverse effects , Sodium Selenite/adverse effects
3.
Phytomedicine ; 17(14): 1133-9, 2010 Dec 01.
Article in English | MEDLINE | ID: mdl-20576410

ABSTRACT

The activity of Uncaria tomentosa preparations on cancer cells was studied using in vitro and in vivo models. IC (50) values were calculated for preparations with different quantitative and qualitative oxindole alkaloid composition: B/W(37) --bark extracted in water at 37 °C, B/W(b)--bark extracted in boiling water, B/50E(37) --bark extracted in 50% ethanol at 37 °C, B/E(b)--bark extracted in boiling 96% ethanol, B/96E(37) --bark extracted in 96% ethanol at 37 °C and B/SRT--bark extracted in water and dichloromethane. Generally, the results obtained showed a high correlation between the total oxindole alkaloid content (from 0.43% to 50.40% d.m.) and the antiproliferative activity of the preparations (IC(50) from >1000 µg/ml to 23.57 µg/ml). B/96E(37) and B/SRT were the most cytotoxic preparations, whereas the lowest toxicity was observed for B/W(37). B/96E(37) were shown to be active against Lewis lung carcinoma (LL/2) [IC(50) =25.06 µg/ml], cervical carcinoma (KB) [IC(50) =35.69 µg/ml] and colon adenocarcinoma (SW707) [IC(50) =49.06 µg/ml]. B/SRT was especially effective in inhibiting proliferation of cervical carcinoma (KB) [IC(50) =23.57 µg/ml], breast carcinoma (MCF-7) [IC(50) =29.86 µg/ml] and lung carcinoma (A-549) [IC(50) =40.03 µg/ml]. Further animal studies on mice bearing Lewis lung carcinoma showed significant inhibition of tumor growth by B/W(37) administered for 21 days at daily doses of 5 and 0.5 mg (p=0.0009). There were no significant changes in the cell cycles of tumor cells with the exception of cell decrease at the G2/M phase after the administration of B/96E(37) at a daily dose of 0.5 mg and the G(1)/G(0) cells cycle arrest demonstrated after the B/SRT therapy at a daily-dose of 0.05 mg. All tested preparations were non-toxic and well tolerated.


Subject(s)
Alkaloids/therapeutic use , Antineoplastic Agents, Phytogenic/therapeutic use , Cat's Claw/chemistry , Indoles/therapeutic use , Neoplasms/drug therapy , Plant Extracts/therapeutic use , Alkaloids/pharmacology , Animals , Antineoplastic Agents, Phytogenic/pharmacology , Cell Cycle/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , Female , Humans , Indoles/pharmacology , Male , Mice , Mice, Inbred C57BL , Oxindoles , Phytotherapy , Plant Bark , Plant Extracts/pharmacology
4.
J Ethnopharmacol ; 121(1): 140-7, 2009 Jan 12.
Article in English | MEDLINE | ID: mdl-19027059

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Uncaria tomentosa (Willd.) DC. is the most popular Peruvian plant, used in folk medicine for different purposes. It contains thousands of active compounds with great content of alkaloids. AIM OF STUDY: Two different fractions of Alkaloid-Rich and Alkaloid-Free were researched on chromosome morphology, mitotic activity and phases indexes. MATERIALS AND METHODS: Cells of Allium Test (meristematic cells of root tips) were incubated up to 24h in different concentrations of Alkaloid-Free and Alkaloid-Rich fraction obtained from Uncaria tomentosa bark followed by 48 h of postincubation in water. The chromosome morphology was analyzed and the content of mitotic and phase indexes were done. Individual compounds, oxindole alkaloids, phenolic compounds and sugars were determined. RESULTS: In Alkaloid-Rich and Alkaloid-Free fractions (different in chemical composition) we observed condensation and contraction of chromosomes (more in Alkaloid-Rich) with retardation and/or inhibition of mitoses and changed mitotic phases. Postincubation reversed results in the highest concentration which was lethal (in mostly Alkaloid-Rich fraction). CONCLUSIONS: Our studies indicate that different action can depend on different groups of active compounds in a preparation either containing alkaloids or not. Other fraction analysis may be useful in the future.


Subject(s)
Alkaloids/pharmacology , Antimitotic Agents/pharmacology , Cat's Claw/chemistry , Cell Nucleus/drug effects , Chromosomes, Plant/drug effects , Onions/drug effects , Alkaloids/isolation & purification , Antimitotic Agents/isolation & purification , Cell Nucleus/genetics , Cell Nucleus/metabolism , Chromosomes, Plant/ultrastructure , Meristem/drug effects , Meristem/genetics , Metaphase/drug effects , Mitotic Index , Onions/genetics , Plant Bark , Plant Extracts/chemistry , Plant Extracts/pharmacology
5.
J Ethnopharmacol ; 107(2): 211-21, 2006 Sep 19.
Article in English | MEDLINE | ID: mdl-16793229

ABSTRACT

The influence of water extract of Uncaria tomentosa (Willd.) DC bark on the meristematic cells of the root tips of Allium cepa L., e.g. cells of Allium Test, was investigated. The experiment was carried out in two variants: (1) continuous incubation at different concentrations (2, 4, 8 and 16 mg/ml) of the extract for 3, 6, 12, 24, 48 and 72h; and (2) 24-h incubation in three concentrations of the extract (4, 8 or 16 mg/ml), followed by post-incubation in distilled water for 3, 6, 12, 24 and 48h. During the continuous incubation, the mitotic activity was reduced (2 and 4 mg/ml) or totally inhibited (8 and 16 mg/ml), depending on the concentration of the extract. All the concentrations resulted in gradual reduction of the mitotic activity. In the concentration of 2 mg/ml, the mitotic activity reached its lowest value after 12h (2 mg/ml) and after 24h in 4 mg/ml, followed by spontaneous intensification of divisions during further incubation. Instead, in higher concentrations of the extracts (8 and 16 mg/ml), the mitotic activity was totally inhibited within 24h and did not resume even after 72h. Incubation caused changes in the phase index, mainly as an increase in the number of prophases. After 24h of incubation, in all phases, condensation and contraction of chromosomes were observed. During post-incubation, divisions resumed in all concentrations, reaching even higher values than the control. Cytometric analysis showed that the extract caused inhibition of the cell cycle at the border between gap(2) and beginning of mitosis (G(2)/M).


Subject(s)
Antimitotic Agents/pharmacology , Cat's Claw/chemistry , Cell Nucleus/drug effects , Chromosomes, Plant/drug effects , DNA, Plant/metabolism , Onions/drug effects , Antimitotic Agents/isolation & purification , Cell Nucleus/genetics , Cell Nucleus/metabolism , Chromosomes, Plant/ultrastructure , Meristem/drug effects , Meristem/genetics , Meristem/metabolism , Metaphase/drug effects , Mitotic Index , Onions/genetics , Onions/metabolism , Plant Bark/chemistry , Plant Extracts/isolation & purification , Plant Extracts/pharmacology
6.
J Ethnopharmacol ; 103(1): 43-52, 2006 Jan 03.
Article in English | MEDLINE | ID: mdl-16169692

ABSTRACT

Rhodiola rosea is a medicinal plant having stimulating and adaptogenic properties, and some reports also indicate its anticancer and antimutagenic effect. However, the mechanism of its anticancer effect is unknown as there have been no cytological studies regarding cytostatics, cell cycle, induction of apoptosis or the mitotic activity of healthy and cancerous cells. In the present paper, those parameters were investigated using HL-60 cells, with flow cytometry and fluorescence microscopy. It has been found that the extract of Rhodiola rosea rhizomes inhibits division of HL-60 cells, which is preceded by an accumulation of cells at the prophase stage. This leads to induction of apoptosis and necrosis in HL-60 cells, and to marked reduction of their survival. The cells enter apoptosis from phase G2/M of the cell cycle. After treatment with the extract, no chromosome aberrations or micronuclei were observed, which indicates the mild action of the extract. The cytostatic and antiproliferative effect of the Rhodiola rosea rhizome extract, and its mild action, raises hope for its use in anticancer therapy by enhancing the effectiveness of cytostatics.


Subject(s)
Antimitotic Agents/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Plant Extracts/pharmacology , Rhodiola , S Phase/drug effects , Chromatography, High Pressure Liquid , Dose-Response Relationship, Drug , HL-60 Cells , Humans , Mitotic Index , Necrosis
7.
Ann Bot ; 90(5): 637-45, 2002 Nov.
Article in English | MEDLINE | ID: mdl-12466105

ABSTRACT

Acclimation of winter oilseed plants in the cold (i.e. at temperatures >0 degrees C) followed by short exposure to sub-lethal freezing temperatures resulted in pronounced ultrastructural changes of leaf epidermal and mesophyll cells. The following major changes were observed upon acclimation at 2 degrees C: increased thickness of cell walls; numerous invaginations of plasma membranes; the appearance of many large vesicles localized in the cytoplasm in close proximity to the central vacuole; the occurrence of abundant populations of microvesicles associated with the endoplasmic reticulum (ER) cisternae or located in the vicinity of dictyosomes; and the occurrence of paramural bodies and myelin-like structures. In addition, large phenolic deposits were observed in the vicinity of the plasma membrane and membrane-bound organelles such as chloroplasts, large vesicles or cytoplasm/tonoplast interfaces. Transient freezing (-5 degrees C for 18 h) of the cold-acclimated leaves led to reversible disorganization of the cytoplasm and to pronounced structural changes of the cellular organelles. Chloroplasts were swollen, with the stroma occupying one half of their volume and the thylakoid system being displaced to the other half. Large phenolic aggregates disappeared but distinct layers of phenolic deposits were associated with mitochondrial membranes and with chloroplast envelopes. In frost-thawed cells recovered at 2 degrees C for 24 h, dictyosomes and dictyosome- or ER-derived small vesicles reappeared in the ribosome-rich cytoplasm. Aberrations in the structure of chloroplasts and mitochondria were less pronounced. Few phenolic deposits were seen as small grains associated with chloroplast envelopes and vesicle membranes. These observations demonstrate that plants undergo different changes in cell ultrastructure depending on whether they are subjected to chilling or freezing temperatures. Results are discussed in relation to membrane recycling and the possible role of phenolics during the first and second stages of plant acclimation at low temperature.


Subject(s)
Brassica napus/chemistry , Brassica napus/ultrastructure , Cold Temperature , Phenols/analysis , Plant Leaves/chemistry , Plant Leaves/ultrastructure , Adaptation, Physiological , Brassica napus/cytology , Environment , Plant Epidermis/chemistry , Plant Epidermis/cytology , Plant Epidermis/ultrastructure , Plant Leaves/cytology
8.
Ann Bot ; 90(2): 269-78, 2002 Aug.
Article in English | MEDLINE | ID: mdl-12197525

ABSTRACT

Light, fluorescence and electron microscopy were used to analyse the structural properties of protoplasts obtained from established suspension culture of Solanum lycopersicoides Dun, composed of meristematic cell aggregates. Four types of protoplasts were distinguished immediately after isolation: (1) mononuclear; (2) polynuclear, (3) anuclear and (4) homogeneous protoplasts. Only mononuclear protoplasts were capable of complete cell wall regeneration and mitotic division. Other types of protoplasts were eliminated during culture. Three phases were distinguished in the developing protoplast culture: (1) the elimination phase during which protoplasts damaged during isolation underwent complete degradation; (2) a phase of intense division during which both mitotic cell division and amitotic nuclear division took place; and (3) a stabilization phase leading to the formation of suspension culture. The cell suspension culture obtained from protoplasts was capable of regenerating diploid plants.


Subject(s)
Diploidy , Solanaceae/genetics , Cell Division/physiology , Cell Nucleus/genetics , Cell Nucleus/physiology , Cell Wall/genetics , Cell Wall/physiology , Culture Techniques , Flow Cytometry , Microscopy, Confocal , Microscopy, Electron , Microscopy, Fluorescence , Protoplasts/cytology , Protoplasts/physiology , Protoplasts/ultrastructure , Solanaceae/growth & development , Solanaceae/ultrastructure
9.
Planta ; 215(3): 371-9, 2002 Jul.
Article in English | MEDLINE | ID: mdl-12111217

ABSTRACT

Tobacco ( Nicotiana tabacum L.) has two major H1 variants (H1A and H1B), which account for over 80% of chromatin linker histones, and four minor variants: H1C, H1D, H1E and H1F. We have shown previously [M. Prymakowska-Bosak et al. (1999) Plant Cell 11:2317-2329] that reversal of the natural proportion of major to minor H1 variants in transgenic tobacco plants results in a characteristic male-sterility phenotype identical to that occurring in many plant species subjected to water deficit at the time of male meiosis. It has been proposed by others that the drought-induced arrest of male gametophyte development is linked to decreased sugar delivery to reproductive tissues. Within the family of angiosperm H1s there is a well-defined class of minor H1 variants named "drought inducible" because some of its members have been shown to be induced by water deficit. We have identified and cloned the tobacco H1C gene, which, based on sequence similarity, represents a "drought-inducible" minor H1 variant. Analysis of the un-translated mRNA and promoter regions of H1C suggests a regulation by sucrose concentration. Antisense silencing of H1C and its close homologue H1D in plants that do not express H1A and H1B does not affect the characteristic H1A(-)/ H1B(-) male-sterility phenotype. Silencing of H1C and H1D also has no effect on growth and development of plants. Our findings demonstrate that H1C and H1D are dispensable for normal growth and development of tobacco, and that the compensatory up-regulation of "drought-inducible" H1s observed in H1A(-)/ H1B(-) plants is not the direct cause of male sterility linked to alterations in H1 variants.


Subject(s)
Histones/genetics , Nicotiana/physiology , Acclimatization , Amino Acid Sequence , Base Sequence , DNA Primers , Disasters , Fertility , Genetic Variation , Germ Cells/physiology , Infertility , Molecular Sequence Data , Phylogeny , Plants, Genetically Modified , Pollen/physiology , Polymerase Chain Reaction , Promoter Regions, Genetic , Sequence Alignment , Sequence Homology, Amino Acid , Spores/physiology , Nicotiana/classification , Nicotiana/genetics
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