A graphene oxide nanosensor enables the co-delivery of aptamer and peptide probes for fluorescence imaging of a cascade reaction in apoptotic signaling

The Analyst ◽  
2018 ◽  
Vol 143 (1) ◽  
pp. 208-214 ◽  
Author(s):  
Chang Liu ◽  
Yan-Lei Hu ◽  
Wen-Jing Deng ◽  
Qing-Shan Pan ◽  
Jin-Tao Yi ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Cytochrome C ◽  
Fluorescence Imaging ◽  
Caspase 3 ◽  
Cascade Reaction ◽  
Apoptotic Signaling ◽  
Cyt C ◽  
Peptide Probes

Cytochrome c (Cyt c) and caspase-3 are the key mediators in apoptotic signaling.

scholarly journals PO-298 Effect of exercise and Siyeshen upon hippocampal cytochrome c and caspase-3 of diabetic rats

2018 ◽  
Vol 1 (5) ◽  
Author(s):  
Dongmei Wang ◽  
Jinming Zhang ◽  
Haibin Liu ◽  
Rongmei Wang
Keyword(s):  
Cytochrome C ◽  
Protein Expression ◽  
Caspase 3 ◽  
Diabetic Control ◽  
Exercise Group ◽  
Diabetic Rats ◽  
Control Group ◽  
Diabetic Control Group ◽  
Mrna And Protein Expression ◽  
Cyt C

Objective To observe the effects of aerobic exercise and Siyeshen water extract on cytochrome c (Cyt c) and caspase-3 in hippocampus of diabetic rats and to explore the possible mechanism of improving diabetes. Methods Healthy male Wister rats fed with high fat and high sugar and combined with streptozotocin to establish type II diabetes model. They were randomly divided into 4 groups: diabetic control group, exercise group, Siyeshen group and exercise+Siyeshen group, and another normal control group, with 6 rats in each group. After aerobic exercise (15m/min, 5°slope, 60min, every other day) or/and Siyeshen (200mg/kg) gastrointestinal administration for 8w, the expression of Cyt c and caspase-3 in hippocampus of each group were detected by immunoblotting, and mRNA expressions were detected by RT-PCR. Results Compared with the normal control group, the mRNA and protein expressions of Cyt c and caspase-3 in the hippocampus of the diabetic control group were significantly increased (P<0.05). Compared with the diabetic control group, the blood glucose level of exercise group and exercise+ Siyeshen group decreased (P<0.05), the mRNA and protein expression of Cyt c and caspase-3 decreased significantly (P<0.05), but there were no significant changes in the mRNA and protein expression of Cyt c and caspase-3 between Siyeshen group and diabetic control group (P﹥0.05). Conclusions Exercise and exercise combined with Siyeshen can inhibit cytochrome c release and reduce caspase-3 protein expression, which may be related to the improvement of blood glucose levels in diabetic rats.

Higher Expression of Cytochrome-c and Caspase-3 in the Primary Acute Lymphoblastic Leukemic Cells with Negative Expression of VEGF and Its Receptors Than with Positive Expressed Cells under VP16 Induction.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4448-4448 ◽  
Author(s):  
Aibin Liang ◽  
Long-Jun Gu
Keyword(s):  
Flow Cytometry ◽  
Cytochrome C ◽  
Caspase 3 ◽  
De Novo ◽  
Leukemic Cells ◽  
Immunofluorescent Staining ◽  
Leukemia Cells ◽  
Cyt C ◽  
Primary Leukemia

Abstract Flow cytometry and immunofluorescent staining were employed to examine the apoptosis signals, cytochrome-c and Caspase-3, in the primary leukemia cells with or without expression of VEGF and its receptors and normal healthy donor’s lymphocytes. We found that primary leukemic cells and normal lymphocytes didn’t secrete Cyt-c from mitochondria and no caspase-3 activation without the treatment of VP16. Instead, after incubation with VP16 (30mg/L) for 8 hours, normal lymphocytes and most of de novo leukemia cells secreted Cyt-c into cytosol from mitochondria and had the activation of caspase-3 in the cellular plasma. However, primary leukemia blasts with higher expression of VEGF and its receptors revealed lower secretion of Cyt-C and down-regulated activation of Caspase-3. Results showed that Cyt-c secretion and Caspase-3 activation were prone to occur in the VEGF negatively expressed blasts than positively expressed cells. This indicates that lower secretion of Cyt-c from mitochondria and inactivation of Caspase-3 are two of the pivotal mechanisms of anti-apoptosis for refractory leukemia clone with highly expressed VEGF and its receptors.

scholarly journals Bcr-Abl Exerts Its Antiapoptotic Effect Against Diverse Apoptotic Stimuli Through Blockage of Mitochondrial Release of Cytochrome C and Activation of Caspase-3

Blood ◽  
1998 ◽  
Vol 91 (5) ◽  
pp. 1700-1705 ◽  
Author(s):  
Gustavo P. Amarante-Mendes ◽  
Caryn Naekyung Kim ◽  
Linda Liu ◽  
Yue Huang ◽  
Charles L. Perkins ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Dna Fragmentation ◽  
Caspase 3 ◽  
K562 Cells ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
High Dose ◽  
Antiapoptotic Effect ◽  
Endogenous Expression ◽  
Cyt C

Abstract Bcr-Abl expression in leukemic cells is known to exert a potent effect against apoptosis due to antileukemic drugs, but its mechanism has not been elucidated. Recent reports have indicated that a variety of apoptotic stimuli cause the preapoptotic mitochondrial release of cytochrome c (cyt c) into cytosol, which mediates the cleavage and activity of caspase-3 involved in the execution of apoptosis. Whether Bcr-Abl exerts its antiapoptotic effect upstream to the cleavage and activation of caspase-3 or acts downstream by blocking the ensuing degradation of substrates resulting in apoptosis, has been the focus of the present studies. In these, we used (1) the human acute myelogenous leukemia (AML) HL-60 cells that are stably transfected with thebcr-abl gene (HL-60/Bcr-Abl) and express p185 Bcr-Abl; and (2) the chronic myelogenous leukemia (CML)-blast crisis K562 cells, which have endogenous expression of p210 Bcr-Abl. Exposure of the control AML HL-60 cells to high-dose Ara-C (HIDAC), etoposide, or sphingoid bases (including C2 ceramide, sphingosine, or sphinganine) caused the accumulation of cyt c in the cytosol, loss of mitochondrial membrane potential (MMP), and increase in the reactive oxygen species (ROS). These preapoptotic events were associated with the cleavage and activity of caspase-3, resulting in the degradation of poly (adenosine diphosphate [ADP]-ribose) polymerase (PARP) and DNA fragmentation factor (DFF), internucleosomal DNA fragmentation, and morphologic features of apoptosis. In contrast, in HL-60/Bcr-Abl and K562 cells, these apoptotic stimuli failed to cause the cytosolic accumulation of cyt c and other associated mitochondrial perturbations, as well as the failure to induce the activation of caspase-3 and apoptosis. While the control HL-60 cells showed high levels of Bcl-2 and barely detectable Bcl-xL, HL-60/Bcr-Abl cells expressed high levels of Bcl-xL and undetectable levels of Bcl-2, a pattern of expression similar to the one in K562 cells. Bax and caspase-3 expressions were not significantly different between HL-60/Bcr-Abl or K562 versus HL-60 cells. These findings indicate that Bcr-Abl expression blocks apoptosis due to diverse apoptotic stimuli upstream by preventing the cytosolic accumulation of cyt c and other preapoptotic mitochondrial perturbations, thereby inhibiting the activation of caspase-3 and execution of apoptosis.

Apoptotic signaling induces hyperpermeability following hemorrhagic shock

2007 ◽  
Vol 292 (6) ◽  
pp. H3179-H3189 ◽  
Author(s):  
Ed W. Childs ◽  
Binu Tharakan ◽  
Felicia A. Hunter ◽  
John H. Tinsley ◽  
Xiaobo Cao
Keyword(s):  
Cytochrome C ◽  
Hemorrhagic Shock ◽  
Caspase 3 ◽  
Mitochondrial Cytochrome ◽  
Intrinsic Apoptotic Pathway ◽  
Intrinsic Pathway ◽  
Apoptotic Pathway ◽  
Apoptotic Signaling ◽  
Mitochondrial Cytochrome C

Hemorrhagic shock (HS) disrupts the endothelial cell barrier, resulting in microvascular hyperpermeability. Recent studies have also demonstrated that activation of the apoptotic signaling cascade is involved in endothelial dysfunction, which may result in hyperpermeability. Here we report involvement of the mitochondrial “intrinsic” pathway in microvascular hyperpermeability following HS in rats. HS resulted in the activation of the mitochondrial intrinsic pathway, as is evident from an increase in the proapoptotic Bcl-2 family member BAK, release of mitochondrial cytochrome c into the cytoplasm, and activation of caspase-3. This, along with the in vivo transfection of the proapoptotic peptide BAK (BH3), resulted in hyperpermeability (as visualized by intravital microscopy), release of mitochondrial cytochrome c into the cytoplasm, and activation of caspase-3. Conversely, transfection of the BAK (BH3) mutant had no effect on hyperpermeability. Together, these results demonstrate involvement of the mitochondrial intrinsic apoptotic pathway in HS-induced hyperpermeability and that the attenuation of this pathway may provide an alternative strategy in preserving vascular barrier integrity.

Angiopoietin-1 inhibits intrinsic apoptotic signaling and vascular hyperpermeability following hemorrhagic shock

2008 ◽  
Vol 294 (5) ◽  
pp. H2285-H2295 ◽  
Author(s):  
Ed W. Childs ◽  
Binu Tharakan ◽  
Nickolas Byrge ◽  
John H. Tinsley ◽  
Felicia A. Hunter ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Hemorrhagic Shock ◽  
Caspase 3 ◽  
Microvascular Endothelial Cell ◽  
Cytochrome C Release ◽  
Apoptotic Signaling ◽  
Microvascular Endothelial ◽  
Lung Microvascular Endothelial Cell ◽  
Angiopoietin 1

Studies from our laboratory demonstrated the involvement of intrinsic apoptotic signaling in hyperpermeability following hemorrhagic shock (HS). Angiopoietin 1 (Ang-1), a potent inhibitor of hyperpermeability, was recently shown to inhibit apoptosis. The purpose of our study was to determine the effectiveness of Ang-1 in attenuating HS-induced hyperpermeability and its relationship to apoptotic signaling. HS was induced in rats by withdrawing blood to reduce the mean arterial pressure to 40 mmHg for 1 h, followed by reperfusion. Mesenteric postcapillary venules were examined for changes in hyperpermeability by intravital microscopy. Mitochondrial release of second mitochondrial derived activator of caspases (smac) and cytochrome c were determined by Western blot and ELISA, respectively. Caspase-3 activity was determined by fluorometric assay. Parallel studies were performed in rat lung microvascular endothelial cell (RLMEC) monolayers, utilizing HS serum and the proapoptotic Bcl-2 homologous antagonist/killer [BAK (BH3)] peptide as inducers of hyperpermeability. In rats, Ang-1 (200 ng/ml) attenuated HS-induced hyperpermeability versus the HS group ( P < 0.05). Ang-1 prevented HS-induced collapse of mitochondrial transmembrane potential (ΔΨm), smac and cytochrome c release, and caspase-3 activity ( P < 0.05). In RLMEC monolayers, HS serum and BAK (BH3) peptide both induced hyperpermeability that was inhibited by Ang-1 ( P < 0.05). Ang-1 attenuated HS and BAK (BH3) peptide-induced collapse of ΔΨm, smac release, cytochrome c release, activation of caspase-3, and vascular hyperpermeability. In vivo, BAK (BH3) induced vascular hyperpermeability that was attenuated by Ang-1 ( P < 0.05). These findings suggest that Ang-1's role in maintaining microvascular endothelial barrier integrity involves the intrinsic apoptotic signaling cascade.

Abstract WP124: Targeting Prodeath Signaling Upstream Of Mitochondria Damage-cathepsins In Astrocytes Contributes To Neuroprotection Against Cerebral Ischmia

Stroke ◽  
2013 ◽  
Vol 44 (suppl_1) ◽  
Author(s):  
Huiling Zhang ◽  
Min Xu ◽  
Lei Yang ◽  
Katunuma Nobuhiko ◽  
Kazumi Ishidoh
Keyword(s):  
Cathepsin B ◽  
Caspase 3 ◽  
Infarct Volume ◽  
Cathepsin L ◽  
Glucose Deprivation ◽  
Selective Inhibitor ◽  
Neurological Deficits ◽  
Apoptotic Signaling ◽  
Cultured Astrocytes ◽  
Cyt C

The roles of cathepsins in the ischemic astrocytic injury remain unclear. Here we test the hypothesis that the activation of cathepsin B and cathepsin L contribute to astrocytic injury via inhibiting cathepsins-mitochondrial apoptotic signaling patheways during acute focal ischemia and oxygen-glucose deprivation (OGD). In a rat model of pMCAO, CA-074Me, a selective inhibitor of cathepsin B, or Clik148, a selective inhibitor of cathepsin L, reduced the infarct volume, improved the neurological deficits and increased the expression of MAP2 and GFAP at 24 h after ischemia. In OGD-induced primary cultured astrocytes injury, CA-074Me or Clik148 significantly decreased the LDH leakage and increased the number of astrocytes in a dose-dependent fasion, and also increased the expression of GFAP. Immunofluorescence showed that the dots distribution of cathepsin L or cathepsin B was seen in astrocytes in the ischemic cortex of sham, while the diffussion distribution of cathepsin L or B was seen in astrocytes in the ischemic cortex of ischemic control rats at 3 h or 6 h after ischemia, suggesting the release of cathepsin L or cathepsin B from lysosome to cytosol after ischemia and the activation of cathepsin L or cathepsin B in ischemic astrocytes. pMCAO also induced the activation of caspase-3 in ischemic astrocytes at 12 h after iscemia. CA-074Me or Clik148 reversed pMCAO-induced activation of cathepsin B or cathepsin L and caspase-3 in ischemic astrocytes. Western Blot analysis showed that OGD induced an increase in activated cathepsin B or cathepsin L, tBid and Caspase-3, reduced Cyt-c in mitochondria and increased Cyt-c in cytoplasm in astrocytes. CA-074Me or Clik148 blocked OGD-induced an increase in activated cathepsin B or cathepsin L, tBid and caspase-3, and a reduction in mitochondria Cyt-c and an increase in cytoplasm Cyt-c. The current findings provide the first evidence that targeting prodeath signaling upstream of mitochondria damage-cathepsins in astrocytes contributes to neuroprotection against cerebral ischmia via blocking the activation of cathepsins-caspase apoptotic signaling pathway in astrocytes.

scholarly journals Bcr-Abl Exerts Its Antiapoptotic Effect Against Diverse Apoptotic Stimuli Through Blockage of Mitochondrial Release of Cytochrome C and Activation of Caspase-3

Blood ◽  
1998 ◽  
Vol 91 (5) ◽  
pp. 1700-1705 ◽  
Author(s):  
Gustavo P. Amarante-Mendes ◽  
Caryn Naekyung Kim ◽  
Linda Liu ◽  
Yue Huang ◽  
Charles L. Perkins ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Dna Fragmentation ◽  
Caspase 3 ◽  
K562 Cells ◽  
Myelogenous Leukemia ◽  
Leukemic Cells ◽  
High Dose ◽  
Antiapoptotic Effect ◽  
Endogenous Expression ◽  
Cyt C

Bcr-Abl expression in leukemic cells is known to exert a potent effect against apoptosis due to antileukemic drugs, but its mechanism has not been elucidated. Recent reports have indicated that a variety of apoptotic stimuli cause the preapoptotic mitochondrial release of cytochrome c (cyt c) into cytosol, which mediates the cleavage and activity of caspase-3 involved in the execution of apoptosis. Whether Bcr-Abl exerts its antiapoptotic effect upstream to the cleavage and activation of caspase-3 or acts downstream by blocking the ensuing degradation of substrates resulting in apoptosis, has been the focus of the present studies. In these, we used (1) the human acute myelogenous leukemia (AML) HL-60 cells that are stably transfected with thebcr-abl gene (HL-60/Bcr-Abl) and express p185 Bcr-Abl; and (2) the chronic myelogenous leukemia (CML)-blast crisis K562 cells, which have endogenous expression of p210 Bcr-Abl. Exposure of the control AML HL-60 cells to high-dose Ara-C (HIDAC), etoposide, or sphingoid bases (including C2 ceramide, sphingosine, or sphinganine) caused the accumulation of cyt c in the cytosol, loss of mitochondrial membrane potential (MMP), and increase in the reactive oxygen species (ROS). These preapoptotic events were associated with the cleavage and activity of caspase-3, resulting in the degradation of poly (adenosine diphosphate [ADP]-ribose) polymerase (PARP) and DNA fragmentation factor (DFF), internucleosomal DNA fragmentation, and morphologic features of apoptosis. In contrast, in HL-60/Bcr-Abl and K562 cells, these apoptotic stimuli failed to cause the cytosolic accumulation of cyt c and other associated mitochondrial perturbations, as well as the failure to induce the activation of caspase-3 and apoptosis. While the control HL-60 cells showed high levels of Bcl-2 and barely detectable Bcl-xL, HL-60/Bcr-Abl cells expressed high levels of Bcl-xL and undetectable levels of Bcl-2, a pattern of expression similar to the one in K562 cells. Bax and caspase-3 expressions were not significantly different between HL-60/Bcr-Abl or K562 versus HL-60 cells. These findings indicate that Bcr-Abl expression blocks apoptosis due to diverse apoptotic stimuli upstream by preventing the cytosolic accumulation of cyt c and other preapoptotic mitochondrial perturbations, thereby inhibiting the activation of caspase-3 and execution of apoptosis.

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