scholarly journals Beclin 1 Interacts With Active Caspase-3 and Bax in Oocytes From Atretic Follicles in the Rat Ovary

2019 ◽  
Vol 67 (12) ◽  
pp. 873-889 ◽  
Author(s):  
María L. Escobar ◽  
Olga M. Echeverria ◽  
Sebastián Palacios-Martínez ◽  
Silvia Juárez-Chavero ◽  
Luis Sánchez-Sánchez ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Beclin 1 ◽  
Follicular Atresia ◽  
Different Types ◽  
Old Rats ◽  
Rat Ovary ◽  
Apoptotic Proteins ◽  
Autophagic Proteins ◽  
Active Caspase

Oocyte cell death is a normal process in the mammalian ovary during follicular growth. Recent reports have demonstrated the presence of pro-apoptotic and pro-autophagic proteins during oocyte elimination. The goal of this study was to identify the interactions between proteins involved in different types of programmed cell death in the same oocyte during follicular atresia. We evaluated the presence of Beclin 1 and its interaction with the pro-apoptotic proteins active caspase-3, Bax, and Bak by means of histochemical observations, ultrastructural immunodetection, and immunoprecipitation techniques in ovaries from prepubertal (28- and 33-day-old), juvenile (40-day-old), and young adult (90-day-old) rats. In this study, we identified that oocyte elimination occurred with a high quantity of pro-autophagic protein Beclin 1 and increased the presence of the pro-apoptotic proteins active caspase-3, Bax, and Bak. Conversely, the antiapoptotic protein Bcl-2 was reduced in oocytes from atretic follicles. In addition, Beclin 1 was shown to interact with active caspase-3 and Bax. Our results suggest that the increase in Beclin 1 is directly related to the rise of pro-apoptotic proteins, which could promote the apoptotic process during oocyte elimination.

Abstract WP62: Enhanced Neuroprotection of Normobaric Oxygenation with Ethanol Conferred by Apoptosis Reduction in Acute Ischemic Stroke

Stroke ◽  
2013 ◽  
Vol 44 (suppl_1) ◽  
Author(s):  
Sweena Parmar ◽  
Xiaokun Geng ◽  
Changya Peng ◽  
Murali Guthikonda ◽  
Yuchuan Ding
Keyword(s):  
Cell Death ◽  
Ischemic Stroke ◽  
Acute Ischemic Stroke ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Neuroprotective Effect ◽  
Apoptotic Cell Death ◽  
Ethanol Administration ◽  
Sprague Dawley ◽  
Apoptotic Proteins

Objectives: Normobaric oxygenation (NBO) has been shown to provide neuroprotection in vivo and in vitro . Yet, a recent Phase 2 clinical trial investigating NBO therapy in acute ischemic stroke was terminated due to questionable therapeutic benefit. NBO therapy alone may be insufficient to produce improved outcomes. In our recent study, we demonstrated a strong neuroprotective effect of ethanol at a dose of 1.5 g/kg (equivalent to the human legal driving limit). In this study, we sought to identify whether low-dose ethanol administration enhances the neuroprotection offered by NBO and whether combined administration of NBO with ethanol is associated with reduced apoptosis. Methods: Sprague-Dawley rats were subjected to right middle cerebral artery occlusion (MCAO) for 2 h, followed by reperfusion. Ischemic animals received either an intraperitoneal injection of 1.0 g/kg ethanol, 2 h of 100% NBO, or both ethanol and NBO. The Cell Death Detection ELISA Assay (Roche) was performed to determine apoptotic cell death at 24 h after reperfusion. Levels of pro-apoptotic (Caspase-3, Bcl-2-associated X-BAX, and Apoptosis-Inducing Factor-AIF) and anti-apoptotic proteins (Bcl-2 and Bcl-xL) were determined by Western blot analysis at 3 and 24 h after reperfusion. Results: As expected, untreated ischemic rats had the highest apoptotic cell death. Combined NBO/ethanol therapy decreased cell death by 48%, as compared to 29% with ethanol and 22% with NBO. Similarly, combined NBO/ethanol therapy promoted the greatest expression of anti-apoptotic factors and the lowest expression of pro-apoptotic proteins at 3 h after reperfusion. This effect was maintained at 24 h and even more pronounced for AIF and Caspase-3. Conclusions: Given singularly, NBO and ethanol improved the degree of cell death, decreased the expression of pro-apoptotic proteins, and increased the expression of anti-apoptotic proteins. Yet, when administered together, their effects largely compounded. These results suggest a synergistic neuroprotection offered by NBO with ethanol, which may be attributed at least in part to their shared role in modulating neuronal apoptosis.

scholarly journals X-linked inhibitor of apoptosis protein and active caspase-3 expression patterns in antral follicles in the sheep ovary

Reproduction ◽  
2011 ◽  
Vol 142 (6) ◽  
pp. 855-867 ◽  
Author(s):  
Hollian R Phillipps ◽  
Ilona C Kokay ◽  
David R Grattan ◽  
Peter R Hurst
Keyword(s):  
Mrna Expression ◽  
Caspase 3 ◽  
Expression Patterns ◽  
Inhibitor Of Apoptosis ◽  
Mrna Levels ◽  
Follicular Atresia ◽  
Inhibitor Of Apoptosis Protein ◽  
Antral Follicles ◽  
Apoptosis Protein ◽  
Active Caspase

X-linked inhibitor of apoptosis protein (XIAP) interacts with caspases to inhibit their activity, thereby providing a potential mechanism for regulation of granulosa cell apoptosis occurring during follicular atresia. The aim of this study was to determine the presence and localization of XIAP mRNA and protein content in the sheep ovary and compare these expression patterns with active caspase-3 protein in the same antral follicles. Romney ewe estrous cycles (n=25) were synchronized with 2–3 Estrumate injections and ovarian tissue collected during the luteal and follicular phases of the cycle. The presence ofXIAPmRNA was confirmed by RT-PCR using laser capture microdissected ovarian cell samples.XIAPmRNA was subsequently localized byin situhybridization histochemistry and XIAP and active caspase-3 protein visualized by immunohistochemistry. In antral follicles extensive XIAP localization was evident in both granulosa and thecal cells. In contrast, mRNA expression was widespread in granulosa cells and only detected in thecal tissue from a small proportion of antral follicles. Active caspase-3 and XIAP comparative expression analysis showed positiveXIAPmRNA expression in all late luteal phase (day 14) follicles, despite varying levels of active caspase-3 protein. A proportion of follicular phase (days 15 and 16) follicles, however, showed an inverse expression relationship at the protein and mRNA levels in both granulosa and thecal tissue, as did XIAP protein in day 14 follicles. These results suggest high XIAP may prevent activation of caspase-3, thereby regulating follicular atresia in antral follicles and could potentially be utilized as a marker of follicular health.

scholarly journals Glutathione Ameliorates Bortezomib (Velcade) -Induced Neuronal Toxicity in N2a and SHSY5Y Cells

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5863-5863
Author(s):  
Jinny Park ◽  
Lalita Subedi ◽  
Su Jung Park ◽  
Oh kyung Lim ◽  
ChanJong Yoo ◽  
...  
Keyword(s):  
Cell Death ◽  
Peripheral Neuropathy ◽  
Cell Survival ◽  
Caspase 3 ◽  
Conflicts Of Interest ◽  
Neuronal Cells ◽  
Lactate Dehydrogenase Activity ◽  
Bortezomib Treatment ◽  
Neuronal Toxicity ◽  
Apoptotic Proteins

Abstract Bortezomib (Velcade, Millennium Pharmaceuticals, Inc.), a potent proteasome inhibitor, is a boronic acid dipeptide that induce cancer cell death. It is an effective drug for both newly diagnosed or relapsed multiple myeloma (MM) patients and is widely used in the treatment of various kinds of lymphomas and other diseases. Peripheral nerve damage is one of the most significant nonhematologic toxicities of bortezomib. The reported incidence of bortezomib-induced peripheral neuropathy (BIPN) ranges from 25 to 75 % . When it occurs, the painful sensory neuropathy can interfere with quality of life (QOL) and with performance of activities of daily living, and it may also adversely affect clinical outcomes by forcing dose modification and/or premature treatment discontinuation. Therefore there are many efforts to improve multiple symptoms associated with BIPN. But there are no reports on amelioration of neurotoxicity which occurred in patients with treatment of bortezomib .Therefore control of this toxicity by the concomitant use of other safe candidates can make it a better option for the cancer therapy with bortezomib. Glutathione (GSH) is a substance produced naturally by the liver and it is also found in fruits, vegetables, and meats. GSH is an important antioxidant and capable of preventing damage to important cellular components caused by reactive oxygen species (ROS) such as free radicals, peroxides, lipid peroxides, and heavy metals. Several small randomized trials have addressed the protective effect of GSH against chemotherapy induced peripheral neuropathy (CIPN )with a platinum agent . In this study we observed that treatment of GSH either in the pre-treatment or in the post treatment inhibited the bortezomib -induced neuronal cell death in both mouse neuronal cells (N2a) as well as dopaminergic neuron of human origin (SHSY5Y). Bortezomib treatment at the concentration of 100 and 200 µg/ml significantly increased the cell death with increased lactate dehydrogenase activity (LDH) and increased apoptotic proteins expressions in the 24 h of treatment of both cells. GSH treatment (1 mg/ml) not only inhibited the bortezomib -induced cell death but also inhibited the LDH activity, suggesting that GSH ameliorates bortezomib -induced neuronal toxicity. A clear activation of the autophagy was also observed in the bortezomib treatment that was lowered by the GSH treatment. In addition to this inhibition of the apoptotic proteins specially Cleaved caspase 3 was observed in the neuronal cells and also the increased Bcl2 and NRF2 protein which are responsible for the cell survival was upregulated with GSH treatment suggesting that GSH increase the cell survival against bortezomib -induced neurotoxicity. Inhibition of the cleaved caspase-3, Bax and LC3 protein are prominent in SHSY5Y cells while activation of the BCL2 and NRF2 is more significant in N2a cells. To elucidate the role of GSH in the development and maintenance of bortezomib -induced neurotoxicity, we will additionally assess ROS and antioxidant enzyme activity levels in the cellular system. This research suggests that GSH ameliorates BIPN and thus can improve the QOL and increase the survival rate of patients treated with bortezomib. Disclosures No relevant conflicts of interest to declare.

scholarly journals Ekspresi Caspase-3 pada Sel Epitel Rongga Mulut (Kb Cell Line) setelah Paparan Ekstrak Kopi

2014 ◽  
Vol 21 (2) ◽  
pp. 122
Author(s):  
Suryani Hutomo ◽  
Yanti Ivana Suryanto ◽  
Heni Susilowati ◽  
Agustinus Rudolf Phym ◽  
Devi Chretella Maheswara
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Caspase 3 ◽  
Positive Control ◽  
Activation Mechanism ◽  
Kb Cells ◽  
Direct Exposure ◽  
Almost All ◽  
Oral Epithelial ◽  
Active Caspase

Kopi adalah minuman yang biasa dikonsumsi oleh masyarakat sehari-hari. Telah diketahui bahwa kopi mengandung kafein seperti yang terdapat juga pada teh dan coklat. Kandungan terbanyak kafein terdapat pada kopi. Kafein mempunyai struktur kimia 1, 3, 7- trimethylxanthine dan merupakan derivat xanthine. Senyawa ini dapat menginduksi kematian sel yang mengarah pada apoptosis, namun mekanisme yang terlibat belum diketahui dengan jelas. Tingginyakonsumsi kopi di dunia yang selalu meningkat mengindikasikan perlunya dilakukan penelitian untuk mengetahui efek kafein pada epitel rongga mulut yang berkontak langsung dengan kafein. Penelitian terdahulu melaporkan bahwaekstrak kopi menyebabkan kerusakan sel yang sebagian besar mengarah pada apoptosis, tetapi mekanismenya belum jelas. Tujuan penelitian ini adalah untuk menganalisis mekanisme kematian sel KB yang diinduksi oleh kafein melaluiaktivasi caspase-3. Sel KB sebagai model epitel oral (5x10⁴ sel) dikultur dalam DMEM menggunakan 24 wells microplate selama 24 jam sebelum perlakuan. Sel selanjutnya dipapar dengan kafein dengan konsentrasi 100 μg/ml, 200 μg/ml, 400 μg/ml dan diinkubasi selama 24 dan 48 jam dalam DMEM. Doxorubicin (0,5625 μg/ml) digunakan sebagai kontrol positif induksi apoptosis. Teknik imunositokimia terhadap caspase-3 dilakukan pada sel setelah dipapar kafeinuntuk mengamati adanya ekspresi caspase-3 sebagai ciri apoptosis. Identifikasi caspase-3 dilakukan menggunakan mikroskop fase kontras. Ekspresi protein caspase-3 terdeteksi pada sitoplasma sel KB. Hasil penelitian ini menunjukkanadanya ekspresi caspase-3 aktif yang ditandai dengan warna cokelat dengan intensitas kuat pada sitoplasma sebagian besar sel setelah dipapar kafein dengan konsentrasi 100 μg/ml dan 200 μg/ml selama 24 jam. Disimpulkan bahwa ekstrak kopi menyebabkan apoptosis sel KB melalui jalur aktivasi caspase-3. ABSTRACT: The Expression of Caspase-3 in Oral Cavity (Kb Cell Line) after Exposure to Coffee Extract. People widely consume coffee in daily meals. It is known there is caffeine found in coffee like it is found in tea and chocolate.Caffeine is found in the greatest amount of coffee. This 1, 3, 7- trimethyl xanthine substance is a derivate of xanthine that is consumed by almost all people in the world. This substance could induce cell death that mainly is apoptosis, but how the mechanism has not been clearly understood. Considering that coffee is widely consumed in the whole world, it is necessary to conduct an experiment to find any possible effect of caffeine to oral epitel that make direct exposure to caffeine. This experiment is targeted to analyze the mechanism of cell death which caused by caffeine through activation of caspase-3. KB cells as oral epithelial model (5x10⁴ sel) were cultured in DMEM using 24 well microplate for 24 hours before treatment. Then caffeine was given with concentration of 100 μg/ml, 200 μg/ml and 400 μg/ml. Cells were then incubated for 24 and 48 hours period in DMEM. Doxorubicin (0,5625 μg/ml) was used as a positive control of apoptosis induction. Immunocytochemistry technique was then done to observe any caspase three expression as amarker for apoptosis. Identification of active caspase-3 was then done using contrast phase microscope. The results showed expression of caspase-3 in KB cells cytoplasm which observed as high intensity of brown colored molecules incell cytoplasm after 100 μg/ml and 200 μg/ml caffeine exposure in 24 hours. It was concluded that coffee extract induce KB cells apoptosis through caspase-3 activation mechanism.

scholarly journals Association of Active Caspase 8 with the Mitochondrial Membrane during Apoptosis: Potential Roles in Cleaving BAP31 and Caspase 3 and Mediating Mitochondrion-Endoplasmic Reticulum Cross Talk in Etoposide-Induced Cell Death

2004 ◽  
Vol 24 (15) ◽  
pp. 6592-6607 ◽  
Author(s):  
Dhyan Chandra ◽  
Grace Choy ◽  
Xiaodi Deng ◽  
Bobby Bhatia ◽  
Peter Daniel ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Death ◽  
Cross Talk ◽  
Membrane Fraction ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
Dominant Negative ◽  
Caspase 8 ◽  
Death Domain ◽  
Active Caspase

ABSTRACT It was recently demonstrated that during apoptosis, active caspase 9 and caspase 3 rapidly accumulate in the mitochondrion-enriched membrane fraction (D. Chandra and D. G. Tang, J. Biol. Chem.278:17408-17420, 2003). We now show that active caspase 8 also becomes associated with the membranes in apoptosis caused by multiple stimuli. In MDA-MB231 breast cancer cells treated with etoposide (VP16), active caspase 8 is detected only in the membrane fraction, which contains both mitochondria and endoplasmic reticulum (ER), as revealed by fractionation studies. Immunofluorescence microscopy, however, shows that procaspase 8 and active caspase 8 predominantly colocalize with the mitochondria. Biochemical analysis demonstrates that both procaspase 8 and active caspase 8 are localized mainly on the outer mitochondrial membrane (OMM) as integral proteins. Functional analyses with dominant-negative mutants, small interfering RNAs, peptide inhibitors, and Fas-associated death domain (FADD)- and caspase 8-deficient Jurkat T cells establish that the mitochondrion-localized active caspase 8 results mainly from the FADD-dependent and tumor necrosis factor receptor-associated death domain-dependent mechanisms and that caspase 8 activation plays a causal role in VP16-induced caspase 3 activation and cell death. Finally, we present evidence that the OMM-localized active caspase 8 can activate cytosolic caspase 3 and ER-localized BAP31. Cleavage of BAP31 leads to the generation of ER- localized, proapoptotic BAP20, which may mediate mitochondrion-ER cross talk through a Ca2+-dependent mechanism.

scholarly journals Cytokine Effects on Cell Viability and Death of Prostate Carcinoma Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-16 ◽  
Author(s):  
Georgios Chondrogiannis ◽  
Michalis Kastamoulas ◽  
Panagiotis Kanavaros ◽  
Georgios Vartholomatos ◽  
Maria Bai ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Death ◽  
Cell Viability ◽  
Prostate Carcinoma ◽  
Lncap Cell ◽  
Caspase 3 ◽  
Carcinoma Cells ◽  
Lncap Cells ◽  
Phosphatidylinositol 3 Kinase ◽  
Active Caspase

We analyzed the effects of IL-13, IFN-γ, and IL-1βon cell viability and death of LNCaP and PC-3 cells and major signaling pathways involved in these effects. Significant increase of LNCaP cell death (apoptotic and necrotic) and increased levels of active caspase 3 were observed in cells treated with inhibitors of ERK 1/2 (UO126) and p38 (SB203580) prior to IL-1βtreatment in comparison to cells treated with UO126, SB203580, or IL-1βalone. Significant increase of LNCaP but not PC-3 cell death was detected after treatment with LY-294002 (inhibitor of phosphatidylinositol 3-kinase). No significant increase of LNCaP and PC-3 cell death was observed after treatment with SP600125 (inhibitor of JNK), SB203580 (inhibitor of p38), UO126 (inhibitor of ERK 1/2), or BAY 11-7082 (inhibitor of NF-κB). Reduced c-FLIPLexpression was observed in LNCaP cells treated with LY-294002. The significant potentiation of LNCaP cell death by inhibition of ERK 1/2, p38, and PI3-K pathways may provide a rationale for therapeutic approach in androgen-dependent prostate cancer.

Oxidative stress and cell death in the cerebral cortex as a long-term consequence of neonatal hypoglycemia

2016 ◽  
Vol 94 (9) ◽  
pp. 1015-1022 ◽  
Author(s):  
T.R. Anju ◽  
P.R. Akhilraj ◽  
C.S. Paulose
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Cell Death ◽  
Cerebral Cortex ◽  
Real Time ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Neonatal Hypoglycemia ◽  
Old Rats

Neonatal hypoglycemia limits glucose supply to cells leading to long-term consequences in brain function. The present study evaluated antioxidant and cell death factors’ alterations in cerebral cortex of 1-month-old rats exposed to neonatal hypoglycemia. Gene expression studies by real-time PCR were carried out using gene-specific TaqMan probes. Fluorescent dyes were used for immunohistochemistry and nuclear staining and imaged by confocal microscope. Total antioxidant level and expression of antioxidant enzymes — superoxide dismutase (SOD) and gluthathione peroxide (GPx) — mRNA was significantly reduced along with high peroxide level in the cerebral cortex of 1-month-old rats exposed to neonatal hypoglycemia. Real-time PCR analysis showed an upregulation of Bax, caspase 3, and caspase 8 gene expression. Confocal imaging with TOPRO-3 staining and immunohistochemistry with caspase 3 antibody indicated cell death activation. The reduced free radical scavenging capability coupled with the expression of key factors involved in cell death pathway points to the possibility of oxidative stress in the cortex of 1-month-old rats exposed to neonatal hypoglycemia. The observed results indicate the effects of neonatal hypoglycemia in determining the antioxidant capability of cerebral cortex in a later stage of life.

scholarly journals Evaluation of programmed cell death processes on the lens epithelium of older dogs with diabetic and hypermature cataracts

2018 ◽  
Vol 55 (2) ◽  
pp. e133668
Author(s):  
Ana Paula Franco do Amaral Hvenegaard ◽  
Paulo Sergio De Moraes Barros ◽  
Angélica Mendonça Vaz Safatle ◽  
Michelle Barbosa Pereira Braga-Sá ◽  
Luana Vicente Melo ◽  
...  
Keyword(s):  
Cell Death ◽  
Cataract Surgery ◽  
Trypan Blue ◽  
Caspase 3 ◽  
Posterior Capsule Opacification ◽  
Beclin 1 ◽  
Cellular Damage ◽  
Lens Epithelial Cells ◽  
Future Studies ◽  
Proliferation And Differentiation

It is well known that posterior capsule opacification (PCO), one of the most common late postoperative complications of cataract surgery, is mainly caused by proliferation and differentiation of remaining lens epithelial cells (LECs) on the posterior lens capsule. Many authors suggest that alterations induced by the pathophysiology of cataracts, its metabolism and the use of 0.1% trypan blue (TB) must cause some degree of cellular damage on these cells, wicht would help to prevent and/or reduce the incidence of PCO after cataract surgery in humans. Therefore, the aim of this study was to evaluate the expression of cell death markers on LECs of older dogs with diabetic and hypermature cataracts, after capsulorhexis, both using 0.1% TB. Twenty samples collected from 13 dogs of different breeds, with ages varying from 8 to 12 years-old, with diabetic and hypermature cataracts, which had been subjected to phacoemulsification surgery (Phaco) using 0.1% TB for staining were studied. Animals were classified as dogs with diabetic (DC) and hypermature cataracts (HC), and expression of molecular markers for apoptosis and autophagy (caspase-3 and beclin-1) on LECs were obtained by immunofluorescence technique. The expression of caspase-3 and beclin-1 was observed in every studied sample and did not differ between groups. In conclusion, our findings suggest that apoptosis and autophagy processes occur to LECs in older dogs presenting diabetic and hypermature cataracts after Phaco utilizing 0.1% TB. Our results may be helpful to future studies of PCO in post-phacoemulsification surgery patients.

Prenatally administered dexamethasone impairs folliculogenesis in spiny mouse offspring

2016 ◽  
Vol 28 (7) ◽  
pp. 1038 ◽  
Author(s):  
Monika Hułas-Stasiak ◽  
Piotr Dobrowolski ◽  
Ewa Tomaszewska
Keyword(s):  
Caspase 3 ◽  
Follicular Development ◽  
Treated Group ◽  
Spiny Mouse ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Follicular Atresia ◽  
Dexamethasone Treatment ◽  
Primordial Follicles ◽  
Acomys Cahirinus ◽  
Active Caspase

This study was designed to determine whether prenatal dexamethasone treatment has an effect on follicular development and atresia in the ovary of spiny mouse (Acomys cahirinus) offspring. Dexamethasone (125 µg kg–1 bodyweight per day) was administered to pregnant spiny mice from Day 20 of gestation to parturition. The processes of follicle loss were analysed using classical markers of apoptosis (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling reaction, active caspase-3) and autophagy (Lamp1). The present study indicated that dexamethasone reduced the pool of healthy primordial follicles. Moreover, the oocytes from these follicles showed intensive caspase-3 and Lamp1 staining. Surprisingly, dexamethasone caused an increase in the number of secondary follicles; however, most of these follicles were characterised by extensive degeneration of the oocyte and caspase-3 and Lamp1 labelling. Western-blot analysis indicated that the glucocorticoid receptor as well as apoptosis and autophagy markers were more strongly expressed in the DEX-treated group than in the control. On the basis of these findings, we have concluded that dexamethasone impairs spiny mouse folliculogenesis and enhances follicular atresia through induction of autophagy or combined autophagy and apoptosis.

Sign in / Sign up

Export Citation Format

Share Document