Chelerythrine
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A quaternary benzo[c]phenanthridine alkaloid chelerythrine displays a wide range of biological activities including cytotoxicity to normal and cancer cells. In contrast, less is known about the biological activity of dihydrochelerythrine, a product of chelerythrine reduction. We examined the cytotoxicity of chelerythrine and dihydrochelerythrine in human promyelocytic leukemia HL-60 cells. After 4h of treatment, chelerythrine induced a dose-dependent decrease in the cell viability with IC50 of 2.6 microM as shown by MTT reduction assay. Dihydrochelerythrine appeared to be less cytotoxic since the viability of cells exposed to 20 microM dihydrochelerythrine for 24h was reduced only to 53%. Decrease in the viability induced by both alkaloids was accompanied by apoptotic events including the dissipation of mitochondrial membrane potential, activation of caspase-9 and -3, and appearance of cells with sub-G1 DNA. Moreover, chelerythrine, but not dihydrochelerythrine, elevated the activity of caspase-8. A dose-dependent induction of apoptosis and necrosis by chelerythrine and dihydrochelerythrine was confirmed by annexin V/propidium iodide dual staining flow cytometry. Besides, both alkaloids were found to induce accumulation of HL-60 cells in G1 phase of the cell cycle. We conclude that both chelerythrine and dihydrochelerythrine affect cell cycle distribution, activate mitochondrial apoptotic pathway, and induce apoptosis and necrosis in HL-60 cells.
- MeSH
- akutní promyelocytární leukemie farmakoterapie patologie MeSH
- alkaloidy aplikace a dávkování farmakologie MeSH
- apoptóza účinky léků MeSH
- benzofenantridiny aplikace a dávkování farmakologie MeSH
- buněčná smrt účinky léků MeSH
- časové faktory MeSH
- DNA nádorová metabolismus účinky léků MeSH
- financování organizované MeSH
- G1 fáze účinky léků MeSH
- HL-60 buňky MeSH
- inhibiční koncentrace 50 MeSH
- kaspasy metabolismus účinky léků MeSH
- lidé MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- protinádorové látky aplikace a dávkování farmakologie MeSH
- viabilita buněk účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- alkaloidy farmakologie MeSH
- antiinfekční látky farmakologie MeSH
- apoptóza účinky záření MeSH
- biotransformace MeSH
- fenantridiny farmakologie MeSH
- finanční podpora výzkumu jako téma MeSH
- isochinoliny farmakologie chemie toxicita MeSH
- kometový test MeSH
- poškození DNA MeSH
- techniky in vitro MeSH
- vztahy mezi strukturou a aktivitou MeSH
OBJECTIVES: This review summarizes the involvement of sanguinarine and chelerythrine in cell cycle regulation and cell death in various cell lines. It is focused on their potential in the treatment of cancer. METHODS: We conducted a search of PubMed, ScienceDirect and Medline for papers on the molecular mechanisms of the biological activity of sanguinarine and chelerythrine published mainly from 1995 to 2006. RESULTS AND CONCLUSIONS: Our analysis of the published studies suggested that these alkaloids are not only good candidates for chemotherapeutic regimens but may also contribute to the development of successful immune therapies of some carcinomas due to their apoptotic potential. However, the complete signalling cascade in which sanguinarine and chelerythrine treatment induces apoptotic cell death is not yet understood. Overall, the results of recent studies suggest that sanguinarine and chelerythrine may be useful as agents in the management of cancer.
- MeSH
- alkaloidy farmakologie MeSH
- apoptóza účinky léků MeSH
- benzofenantridiny farmakologie MeSH
- buněčné linie MeSH
- buněčný cyklus účinky léků MeSH
- isochinoliny farmakologie MeSH
- lidé MeSH
- protinádorové látky farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Quaternary benzo[c]phenanthridine alkaloids (QBA) sanguinarine and chelerythrine exhibit a wide spectrum of biological activities whence they are used in dental care products. Recent studies indicated that cytochrome P450 CYP1A attenuates sanguinarine toxicity both in vivo [Williams, M.K., Dalvi, S., Dalvi, R.R., 2000. Influence of 3-methylcholanthrene pretreatment on sanguinarine toxicity in mice. Vet. Hum. Toxicol. 42, 196-198] and in vitro [Vrba, J., Kosina, P., Ulrichová, J., Modrianský, M., 2004. Involvement of cytochrome P450 1A in sanguinarine detoxication. Toxicol. Lett. 151, 375-387]. However, CYP1A converts sanguinarine to the products that form DNA adducts [Stiborová, M., Simánek, V., Frei, E., Hobza, P., Ulrichová, J., 2002. DNA adduct formation from quaternary benzo[c]phenanthridine alkaloids sanguinarine and chelerythrine as revealed by the 32P-postlabeling technique. Chem. Biol. Interact. 140, 231-242]. In our work we examined the effects of sanguinarine and chelerythrine on CYP1A1 expression and catalytic activity in human hepatoma cells-HepG2. Sanguinarine and chelerythrine did not affect basal and dioxin-inducible expression of CYP1A1 mRNA and protein in HepG2 cells. The enzymatic activity of CYP1A1 was assessed by the fluorescent measurement of 7-ethyxoresorufin-O-deethylase (EROD) activity. We observed a slight decrease of dioxin-induced EROD activity in HepG2 cells by sanguinarine and chelerythrine. This decrease was attributed to the inhibition of CYP1A1 catalytic activity, as revealed by enzyme kinetic studies on recombinant CYP1A1 protein. The IC50 values for the inhibition of CYP1A1 by sanguinarine and chelerythrine were 2.1 and 1.9muM, respectively. In conclusion, albeit the CYP1A modulates QBA cytotoxicity and genotoxicity, the QBA themselves do not affect CYP1A1 expression. The data indicate that studied alkaloids do not have specific cellular target and their biological effects are rather pleiotropic.
- MeSH
- alkaloidy farmakologie MeSH
- benzofenantridiny MeSH
- cytochrom P-450 CYP1A1 antagonisté a inhibitory biosyntéza MeSH
- cytochrom P-450 CYP1A2 biosyntéza MeSH
- fenantridiny farmakologie MeSH
- financování organizované MeSH
- hepatocelulární karcinom enzymologie MeSH
- inhibitory cytochromu P450 CYP1A2 MeSH
- inhibitory enzymů farmakologie MeSH
- isochinoliny MeSH
- katalýza MeSH
- lidé MeSH
- messenger RNA biosyntéza MeSH
- nádorové buněčné linie MeSH
- nádory jater enzymologie MeSH
- rekombinantní proteiny chemie MeSH
- Check Tag
- lidé MeSH
Costunolide, a natural sesquiterpene lactone, has multiple pharmacological activities such as neuroprotection or induction of apoptosis and eryptosis. However, the effects of costunolide on pro-survival factors and enzymes in human erythrocytes, e.g. glutathione and glucose-6-phosphate dehydrogenase (G6PDH) respectively, have not been studied yet. Our aim was to determine the mechanisms underlying costunolide-induced eryptosis and to reverse this process. Phosphatidylserine exposure was estimated from annexin-V-binding, cell volume from forward scatter in flow cytometry, and intracellular glutathione [GSH]i from high performance liquid chromatography. The oxidized status of intracellular glutathione and enzyme activities were measured by spectrophotometry. Treatment of erythrocytes with costunolide dose-dependently enhanced the percentage of annexin-V-binding cells, decreased the cell volume, depleted [GSH]i and completely inhibited G6PDH activity. The effects of costunolide on annexin-V-binding and cell volume were significantly reversed by pre-treatment of erythrocytes with the specific PKC-α inhibitor chelerythrine. The latter, however, had no effect on costunolide-induced GSH depletion. Costunolide induces eryptosis, depletes [GSH]i and inactivates G6PDH activity. Furthermore, our study reveals an inhibitory effect of chelerythrine on costunolide-induced eryptosis, indicating a relationship between costunolide and PKC-α. In addition, chelerythrine acts independently of the GSH depletion. Understanding the mechanisms of G6PDH inhibition accompanied by GSH depletion should be useful for development of anti-malarial therapeutic strategies or for synthetic lethality-based approaches to escalate oxidative stress in cancer cells for their sensitization to chemotherapy and radiotherapy.
- MeSH
- apoptóza účinky léků MeSH
- benzofenantridiny farmakologie MeSH
- eryptóza účinky léků genetika MeSH
- erytrocyty účinky léků patologie MeSH
- glukosa-6-fosfátdehydrogenasa antagonisté a inhibitory genetika MeSH
- glutathion genetika MeSH
- inhibitory enzymů farmakologie MeSH
- lidé MeSH
- oxidační stres účinky léků MeSH
- proteinkinasa C-alfa antagonisté a inhibitory genetika MeSH
- reaktivní formy kyslíku MeSH
- seskviterpeny farmakologie MeSH
- vápník metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Inhibition of porcine pancreas and human saliva alpha-amylase (EC 3.2.1.1) by sanguinarine and chelerythrine was studied. The inhibition of alpha-amylase was assayed using a biosensor method which utilises a flow system equipped with a peroxide electrode. 250 microM sanguinarine and 250 microM chelerythrine cause complete inhibition of 1.9 nkat alpha-amylase from porcine pancreas. The same concentration of sanguinarine and chelerythrine caused 23.9% and 7.5% inhibition, respectively, of 1.9 nkat alpha-amylase from human saliva. Mixed type and partially reversible inhibition was found for both alpha-amylases treated with either alkaloid.
- MeSH
- alfa-amylasy antagonisté a inhibitory metabolismus MeSH
- alkaloidy farmakologie metabolismus MeSH
- biosenzitivní techniky MeSH
- časové faktory MeSH
- fenantridiny farmakologie metabolismus MeSH
- financování organizované MeSH
- inhibitory enzymů farmakologie MeSH
- kinetika MeSH
- lidé MeSH
- prasata MeSH
- stabilita enzymů MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
The quaternary benzo[c]phenanthridine alkaloid chelerythrine is widely used as an inhibitor of protein kinase C (PKC). However, in biological systems chelerythrine interacts with an array of proteins. In this study, we examined the effects of chelerythrine and sanguinarine on conventional PKCs (cPKCs) and PKC upstream kinase, phosphoinositide-dependent protein kinase 1 (PDK1), under complete inhibition conditions of PKC-dependent oxidative burst. In neutrophil-like HL-60 cells, sanguinarine and chelerythrine inhibited N-formyl-Met-Leu-Phe, phorbol 12-myristate 13-acetate (PMA)-, and A23187-induced oxidative burst with IC(50) values not exceeding 4.6 micromol/L, but the inhibition of PMA-stimulated cPKC activity in intact cells required at least fivefold higher alkaloid concentrations. At concentrations below 10 micromol/L, sanguinarine and chelerythrine prevented phosphorylation of approximately 80 kDa protein and sequestered approximately 60 kDa phosphoprotein in cytosol. Moreover, neither sanguinarine nor chelerythrine impaired PMA-stimulated translocation of autophosphorylated PKCalpha/betaII isoenzymes, but both alkaloids induced dephosphorylation of the turn motif in PKCalpha/betaII. The dephosphorylation did not occur in unstimulated cells and it was not accompanied by PKC degradation. Furthermore, cell treatment with sanguinarine or chelerythrine resulted in phosphorylation of approximately 70 kDa protein by PDK1. We conclude that PKC-dependent cellular events are affected by chelerythrine primarily by multiple protein interactions rather than by inhibition of PKC activity.
- MeSH
- alkaloidy farmakologie chemie MeSH
- benzofenantridiny farmakologie chemie MeSH
- bezbuněčný systém MeSH
- buněčná smrt účinky léků MeSH
- fosforylace účinky léků MeSH
- HL-60 buňky MeSH
- isochinoliny farmakologie chemie MeSH
- izoenzymy metabolismus MeSH
- lidé MeSH
- NADPH-oxidasy metabolismus MeSH
- neutrofily cytologie enzymologie účinky léků MeSH
- protein-serin-threoninkinasy metabolismus MeSH
- proteinkinasa C metabolismus MeSH
- respirační vzplanutí účinky léků MeSH
- substrátová specifita účinky léků MeSH
- transport proteinů účinky léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Quaternary benzo[c]phenanthridine alkaloids sanguinarine and chelerythrine have been used in folk medicine for their wide range of useful properties. One of their major effect is also anti-inflammatory activity, that is not clarified in detail. This study focused on the ability of these alkaloids to modulate the gene expression of pro-inflammatory tumour necrosis factor α (TNF-α), monocyte chemoattractant protein 1 (MCP-1, also known as CCL-2), interleukin (IL)-6, IL-1β and anti-inflammatory cytokines IL-1 receptor antagonist (IL-1RA) and IL-10. The effect of these alkaloids was compared with that of conventional drug prednisone. Human monocyte-derived macrophages were pre-treated with alkaloids or prednisone and inflammatory reaction was induced by lipopolysaccharide. Changes of gene expression at the transcriptional level of mentioned cytokines were measured. In our study mainly affected pro-inflammatory cytokines were CCL-2 and IL-6. Two hours after LPS stimulation, cells influenced by sanguinarine and chelerythrine significantly declined the CCL-2 expression by a factors of 3.5 (p<0.001) and 1.9 (p<0.01); for those treated with prednisone the factor was 5.3 (p<0.001). Eight hours after LPS induction, both alkaloids significantly diminished the CCL-2 expression. The lower expression was found for sanguinarine--lower by a factor of 4.3 than for cells treated with the vehicle (p<0.001). Two hours after LPS stimulation, cells treated with sanguinarine decreased the IL-6 mRNA level by a factor of 3.9 (p<0.001) compared with cells treated with the vehicle. Chelerythrine decreased the level of IL-6 mRNA by a factor of 1.6 (p<0.001). Sanguinarine decreased gene expression of CCL-2 and IL-6 more than chelerythrine and its effect was quite similar to prednisone. Four hours after LPS stimulation, cells pre-treated with sanguinarine exhibited significantly higher expression (a factor of 1.7, p<0.001) of IL-1RA than cells without sanguinarine treatment. Our results help to clarify possible mechanisms of action of these alkaloids in the course of inflammation.
- MeSH
- antiflogistika farmakologie terapeutické užití MeSH
- benzofenantridiny farmakologie terapeutické užití MeSH
- buněčné linie MeSH
- chemokin CCL2 genetika metabolismus MeSH
- exprese genu účinky léků MeSH
- fytoterapie MeSH
- genetická transkripce účinky léků MeSH
- interleukin-6 genetika metabolismus MeSH
- isochinoliny farmakologie terapeutické užití MeSH
- lidé MeSH
- lipopolysacharidy MeSH
- makrofágy účinky léků metabolismus MeSH
- mediátory zánětu metabolismus MeSH
- prednison farmakologie terapeutické užití MeSH
- rostlinné extrakty farmakologie terapeutické užití MeSH
- zánět farmakoterapie genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
