cytochrome-c-t and caffeic-acid-phenethyl-ester

This study aimed to evaluate the protective effects of caffeic acid phenethyl ester (CAPE) and combined CAPE-bevacizumab against oxidative stress induced by hydrogen peroxide (H. ARPE-19 cells were pretreated with 5, 10, and 30 μM CAPE alone and in combination with bevacizumab for 3 h, then exposed to H. Pretreatment of ARPE-19 cells with 30 μM CAPE and combined CAPE-bevacizumab reduced H. CAPE has a protective effect on ARPE-19 cells against oxidative stress, and VEGF protein level and expression can be decreased by incubation with different concentrations of CAPE. These results demonstrate that CAPE suppresses the mitochondria-mediated apoptosis in ARPE-19 cells under oxidative stress. In addition, the use of CAPE in combination with bevacizumab has an additive effect.

Topics: Angiogenesis Inhibitors; Apoptotic Protease-Activating Factor 1; bcl-2-Associated X Protein; Bevacizumab; Caffeic Acids; Caspase 3; Cell Line; Cell Survival; Cytochromes c; Drug Therapy, Combination; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation; Humans; Hydrogen Peroxide; Oxidants; Oxidative Stress; Phenylethyl Alcohol; Proto-Oncogene Proteins c-bcl-2; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Retinal Pigment Epithelium; Vascular Endothelial Growth Factor A

Restenosis after a percutaneous coronary intervention (PCI) during treatment for coronary artery disease is closely related to smooth muscle cell (SMC) proliferation and migration. In this study, we investigated the effects of caffeic acid phenethyl ester (CAPE) and its underlying mechanism on human coronary SMCs (HCSMCs) after platelet-derived growth factor-BB (PDGF-BB) stimulation in vitro.. The results showed that CAPE inhibited proliferation and migration, and induced apoptosis. Concomitantly, CAPE inhibited activation of AKT1, MEK1 and ERK1/2 signaling molecules at 10-60 min after CAPE treatment. As revealed by flow cytometry, DNA fragmentation and TUNEL assay, the cells accumulated at the sub-G(1) phase, and cell apoptosis was observed after 30 and 90 μM CAPE treatment for 72 h. CAPE triggered the release of cytochrome c from mitochondria to cytosol, upregulated the proapoptotic gene Bax and downregulated the antiapoptotic gene Bcl-2. Upregulation of caspase-9 and caspase-3 indicated that CAPE precipitated the mitochondrion-dependent apoptotic signaling pathway.. These results provide a molecular explanation for the antiproliferation, antimigration and proapoptotic effects of CAPE on HCSMCs after PDGF-BB stimulation.

Topics: Apoptosis; Becaplermin; Caffeic Acids; Caspases; Cell Movement; Cell Proliferation; Cells, Cultured; Coronary Vessels; Cytochromes c; Humans; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Phenylethyl Alcohol; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins c-sis

Caffeic acid phenethyl ester (CAPE) is an active component of propolis obtained from honeybee hives and is found to have the following properties: anti-mitogenic, anti-carcinogenic, anti-inflammatory, immunomodulatory, and antioxidant. Recent reports suggest that CAPE also has a neuronal protective property against ischemic injury. Since excitotoxicity may play an important role in ischemia, in this study, we investigated whether CAPE could directly protect neurons against excitotoxic insult. We treated cultured rat cerebellar granule neurons (CGNs) with excitotoxic concentrations of glutamate in the presence or absence of CAPE and found that CAPE markedly protected neurons against glutamate-induced neuronal death in a concentration-dependent fashion. Glutamate-induced CGNs death is associated with time-dependent activation of caspase-3 and phosphorylation of p38, both events of which can be blocked by CAPE. Treating CGNs with specific inhibitors of these two enzymes together exerts a synergistic neuroprotective effect, similar to the neuroprotective effect of CAPE exposure. These results suggest that CAPE is able to block glutamate-induced excitotoxicity by inhibiting phosphorylation of p38 and caspase-3 activation. This finding may further help understanding of the mechanism of glutamate-induced neuronal death and CAPE-induced neuroprotection against excitotoxicity.

Topics: Animals; Animals, Newborn; Caffeic Acids; Caspase 3; Cell Death; Cell Survival; Cells, Cultured; Cerebellum; Cytochromes c; Dose-Response Relationship, Drug; Drug Interactions; Glutamic Acid; Membrane Potentials; Mitochondria; Neurons; Neuroprotective Agents; Neurotoxins; p38 Mitogen-Activated Protein Kinases; Patch-Clamp Techniques; Phenylethyl Alcohol; Phosphorylation; Rats

Caffeic acid phenyl ester (CAPE), a biologically active ingredient of propolis, has several interesting biological properties including antioxidant, anti-inflammatory, antiviral, immunostimulatory, anti-angiogenic, anti-invasive, anti-metastatic and carcinostatic activities. Recently, several groups have reported that CAPE is cytotoxic to tumor cells but not to normal cells. In this study, we investigated the mechanism of CAPE-induced apoptosis in human myeloid leukemia U937 cells. Treatment of U937 cells with CAPE decreased cell viability in a dose-dependent and time-dependent manner. DNA fragmentation assay revealed the typical ladder profile of oligonucleosomal fragments in CAPE-treated U937 cells. In addition, as evidenced by the nuclear DAPI staining experiment, we observed that the nuclear condensation, a typical phenotype of apoptosis, was found in U937 cells treated with 5 microg/ml of CAPE. Therefore, it was suggested that CAPE is a potent agent inducing apoptosis in U937 cells. Apoptotic action of the CAPE was accompanied by release of cytochrome C, reduction of Bcl-2 expression, increase of Bax expression, activation/cleavage of caspase-3 and activation/cleavage of PARP in U937 cells, but not by Fas protein, an initial mediator in the death signaling, or by phospho-eIF2 alpha and CHOP, crucial mediators in ER-mediated apoptosis. From the results, it was concluded that CAPE induces the mitochondria-mediated apoptosis but not death receptors- or ER-mediated apoptosis in U937 cells.

Topics: Apoptosis; Caffeic Acids; Caspase 3; Cell Line, Tumor; Chromatin; Cytochromes c; DNA Fragmentation; Endoplasmic Reticulum; Eukaryotic Initiation Factor-2; fas Receptor; Humans; Leukemia, Myeloid; Mitochondria; Phenylethyl Alcohol; Poly(ADP-ribose) Polymerases; Proto-Oncogene Proteins c-bcl-2; Receptors, Death Domain; Transcription Factor CHOP; U937 Cells

It is an important therapeutic strategy to protect mitochondria from oxidative stress, especially during ischemia-reperfusion. In the present study, an attempt has been made to evaluate the protective effects of caffeic acid phenethyl ester (CAPE) and its related phenolic compounds on mouse brain and liver mitochondria injury induced by in vitro anoxia-reoxygenation. Added before anoxia or reoxygenation, CAPE markedly protected coupled respiration with the decrease in state 4 and the increases in state 3, respiratory control ratio (RCR) and ADP/O ratio in a concentration-dependent manner. CAPE effectively protected mitochondria by inhibiting the mitochondrial membranes fluidity decrease, the lipoperoxidation and the protein carbonylation increase, which indicated its protective action against the mitochondrial oxidative damage. Meanwhile, CAPE blocked the enhanced release of cardiolipin (CL) and cytochrome c (Cyt c). The related phenolic compounds like caffeic acid (CA), ferulic acid (FA) and ethyl ferulate (EF) also had different-degree protective effects. CAPE and CA were more potent than FA and EF. Their structural differences played the key role in their activity levels. These results suggest that CAPE and its related phenolic compounds protect mitochondria mainly correlated to their antioxidative activities and may be of interest for the prevention and therapy of ischemia-reperfusion injuries.

Topics: Anaerobiosis; Animals; Anisotropy; Antioxidants; Brain; Caffeic Acids; Coumaric Acids; Cytochromes c; Dose-Response Relationship, Drug; Male; Mice; Mitochondria; Mitochondria, Liver; Mitochondrial Membranes; Molecular Structure; Oxygen; Phenylethyl Alcohol; Thiobarbituric Acid Reactive Substances

We recently demonstrated that two new prenylflavanones, propolin A and propolin B, isolated and characterized from Taiwanese propolis, induced cytotoxicity effect in human melanoma A2058 cells and shows a strong capability to scavenge free radicals. In this study, propolin A effectively induced a cytotoxic effect on five different cancer cell lines. Similar results were obtained for propolin B. DNA flow cytometric analysis and DNA fragmentation ladder indicated that propolin A and propolin B actively induced apoptosis in A2058 cells. To address the mechanism of the apoptosis effect of propolin A and propolin B, we evaluated the apoptosis-related proteins in A2058 cells. The levels of procaspase-8, Bid, procaspase-3, DFF45, and PARP were decreased in dose- and time course-dependent manners. Furthermore, also found propolin A and propolin B was capable of releasing cytochrome c from mitochondria to cytosol. The findings suggest that propolin A and propolin B may activate a mitochondria-mediated apoptosis pathway. On the other hand, our data show that propolin B inhibitied xanthine oxidase activity more efficiently than propolin A or CAPE. However, CAPE suppressed ROS-induced DNA strand breakage more efficiently than propolin A or propolin B. All these results indicated that propolin A and propolin B may trigger apoptosis of A2058 cells through mitochondria-dependent pathways and also shown that propolin A and propolin B were strong antioxidants.

Topics: Apoptosis; Blotting, Western; Caffeic Acids; Caspase 3; Caspase 8; Cell Cycle; Cell Line, Tumor; Cell Survival; Cytochromes c; DNA Breaks; DNA Fragmentation; DNA, Circular; Dose-Response Relationship, Drug; Flavonoids; Flow Cytometry; HL-60 Cells; Humans; Melanoma; Mitochondria; Molecular Structure; Phenylethyl Alcohol; Propolis; Reactive Oxygen Species; Signal Transduction; Xanthine Oxidase

Although great achievements have been made in elucidating the molecular mechanisms contributing to acute myocardial ischemia/reperfusion (I/R) injury, an effective pharmacological therapy to protect cardiac tissues from serious damage associated with acute myocardial infarction, coronary arterial bypass grafting surgery, or acute coronary syndromes has not been developed. We examined the in vivo cardioprotective effects of caffeic acid phenethyl ester (CAPE), a natural product with potent anti-inflammatory, antitumor, and antioxidant activities. CAPE was systemically delivered to rabbits either 60 min before or 30 min after surgically inducing I/R injury. Infarct dimensions in the area at risk were reduced by >2-fold (P < 0.01) with CAPE treatment at either period. Accordingly, serum levels of normally cytosolic enzymes lactate dehydrogenase, creatine kinase (CK), MB isoenzyme of CK, and cardiac-specific troponin I were markedly reduced in both CAPE treatment groups (P < 0.05) compared with the vehicle-treated control group. CAPE-treated tissues displayed significantly less cell death (P < 0.05), which was in part due to inhibition of p38 mitogen-activated protein kinase activation and reduced DNA fragmentation often associated with caspase 3 activation (P < 0.05). In addition, CAPE directly blocked calcium-induced cytochrome c release from mitochondria. Finally, the levels of inflammatory proteins IL-1beta and TNF-alpha expressed in the area at risk were significantly reduced with CAPE treatment (P < 0.05). These data demonstrate that CAPE has potent cardioprotective effects against I/R injury, which are mediated, at least in part, by the inhibition of inflammatory and cell death responses. Importantly, protection is conferred when CAPE is systemically administered after the onset of ischemia, thus demonstrating potential efficacy in the clinical scenario.

Topics: Acute Disease; Animals; Apoptosis; Blotting, Western; Caffeic Acids; Cardiotonic Agents; Caspases; Creatine Kinase; Cytochromes c; In Situ Nick-End Labeling; Inflammation; Interleukin-1; L-Lactate Dehydrogenase; Male; Mitochondria, Heart; Myocardial Reperfusion Injury; p38 Mitogen-Activated Protein Kinases; Phenylethyl Alcohol; Rabbits; Rats; Rats, Sprague-Dawley; Troponin I; Tumor Necrosis Factor-alpha

We had demonstrated that two prenylflavanones, propolin A and propolin B, isolated and characterized from Taiwanese propolis, induced apoptosis in human melanoma cells and significantly inhibited xanthine oxidase activity. Here, we have isolated a third compound called propolin C. The chemical structure of propolin C has been characterized by NMR and HRMS spectra, and was identical to nymphaeol-A. However, no biological activities of this compound have ever been reported. In the present study, propolin C effectively induced a cytotoxic effect on human melanoma cells, with an IC(50) of about 8.5 microM. DNA flow cytometric analysis indicated that propolin C actively induced apoptosis in human melanoma cells and there is a marked loss of cells from the G2/M phase of the cell cycle. To address the mechanism of the apoptosis effect of propolin C, we evaluated the effect of propolin C on induction of apoptosis-related proteins in human melanoma cells. The levels of procaspase-8, Bid, procaspase-3, and poly(ADP-ribose) polymerase were decreased in dose- or time course-dependent manners. Moreover, propolin C was capable of releasing cytochrome c from mitochondria to cytosol. The findings suggest that propolin C may activate a mitochondria-mediated apoptosis pathway. On other hand, propolin C is a potential antioxidant agent and shows a strong capability to scavenge free radicals and inhibit on xanthine oxidase activity with IC(50) of about 17.0microM. In conclusion, the isolation and characterization of propolin C from bee propolis are described for the first time, and this compound is a powerful inducer of apoptosis in human melanoma cells.

Topics: Antineoplastic Agents; Apoptosis; BH3 Interacting Domain Death Agonist Protein; Caffeic Acids; Carrier Proteins; Caspases; Cell Cycle; Cell Division; Cell Survival; Cytochromes c; DNA Fragmentation; Enzyme Activation; Flavonoids; Free Radicals; Humans; Melanoma; Mitochondria; Phenylethyl Alcohol; Propolis; Xanthine Oxidase

Neonatal hypoxic-ischaemic (HI) brain injury resulting in encephalopathy is a leading cause of morbidity and mortality with no effective treatment. Here we show that caffeic acid phenethyl ester (CAPE), an active component of propolis, administered either before or after an HI insult, significantly prevents HI-induced neonatal rat brain damage in the cortex, hippocampus and thalamus. In addition to blocking HI-induced caspase 3 activation, CAPE also inhibits HI-mediated expression of inducible nitric oxide synthase and caspase 1 in vivo and potently blocks nitric oxide-induced neurotoxicity in vitro. Furthermore, CAPE directly inhibits Ca2+-induced cytochrome c release from isolated brain mitochondria. Thus, CAPE induces neuroprotection against HI-induced neuronal death, possibly by blocking HI-induced inflammation and/or directly inhibiting the HI-induced neuronal death pathway. CAPE may therefore be a novel effective therapy for preventing neonatal HI injury.

Topics: Animals; Animals, Newborn; Caffeic Acids; Calcium; Caspase 3; Caspases; Cells, Cultured; Cytochromes c; Drug Evaluation, Preclinical; Enzyme Activation; Hypoxia-Ischemia, Brain; Mitochondria; Neurons; Neuroprotective Agents; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Phenylethyl Alcohol; Rats; Rats, Sprague-Dawley

Caffeic acid phenethyl ester (CAPE), an active component of propolis, has many biological and pharmacological activities including antioxidant, anti-inflammation, antiviral action, and anticancer effect. Our previous studies showed that CAPE exhibited significant cytotoxicity in oral cancer cells. Herein we further investigated the cytotoxicity potential of CAPE and the mechanism of its action in C6 glioma cells. The data exhibited that C6 glioma cells underwent internucleosomal DNA fragmentation 24 hr after the treatment of CAPE (50 microM). The proportion of C6 glioma cells with hypodiploid nuclei was increased to 24% at 36 hr after the exposure. Further results showed that CAPE induced the release of cytochrome c from mitochondria into cytosol, and the activation of CPP32. CAPE application also enhanced the expression of p53, Bax, and Bak. Finally, the potential signaling components underlying CAPE induction of apoptosis were elucidated. We found that CAPE activated extracellular signal-regulated kinase (ERKs) and p38 mitogen-activated protein kinase (p38 MAPK) in C6 glioma cells. More importantly, p38 kinase formed a complex with p53 after the treatment of CAPE for 0.5 hr. The expression of p53, phospho-serine 15 of p53, and Bax, and inactivate form of CPP32 was suppressed by a pretreatment of a specific p38 MAPK inhibitor, SB203580. The resultant data suggest that p38 MAPK mediated the CAPE-induced p53-dependent apoptosis in C6 glioma cells.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Apoptosis; Caffeic Acids; Caspase 3; Caspases; Cytochromes c; Glioma; Imidazoles; Mitogen-Activated Protein Kinases; p38 Mitogen-Activated Protein Kinases; Phenylethyl Alcohol; Phosphorylation; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerases; Proteins; Proto-Oncogene Proteins c-bcl-2; Pyridines; Rats; Serine; Tumor Cells, Cultured; Tumor Suppressor Protein p53