Anti-apoptotic properties of carbon monoxide in porcine oocyte duringin vitroaging

If fertilization of matured oocyte does not occur, unfertilized oocyte undergoes aging, resulting in a time-dependent reduction of the oocyte’s quality. The aging of porcine oocytes can lead to apoptosis. Carbon monoxide (CO), a signal molecule produced by the heme oxygenase (HO), possesses cytoprotective and anti-apoptotic effects that have been described in somatic cells. However, the effects of CO in oocytes have yet to be investigated. By immunocytochemistry method we detected that both isoforms of heme oxygenase (HO-1 and HO-2) are present in the porcine oocytes. Based on the morphological signs of oocyte aging, it was found that the inhibition of both HO isoforms by Zn-protoporphyrin IX (Zn-PP IX) leads to an increase in the number of apoptotic oocytes and decrease in the number of intact oocytes during aging. Contrarily, the presence of CO donors (CORM-2 or CORM-A1) significantly decrease the number of apoptotic oocytes while increasing the number of intact oocytes. We also determined that CO donors significantly decrease the caspase-3 (CAS-3) activity. Our results suggest that HO/CO contributes to the sustaining viability through regulation of apoptosis duringin vitroaging of porcine oocytes.

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The effect of carbon monoxide on meiotic maturation of porcine oocytes

PeerJ ◽

10.7717/peerj.10636 ◽

2021 ◽

Vol 9 ◽

pp. e10636

Author(s):

David Němeček ◽

Eva Chmelikova ◽

Jaroslav Petr ◽

Tomas Kott ◽

Markéta Sedmíková

Keyword(s):

Oxidative Stress ◽

Carbon Monoxide ◽

Heme Oxygenase ◽

Protein Localization ◽

Somatic Cells ◽

Antioxidant Properties ◽

Protoporphyrin Ix ◽

Antioxidant Effect ◽

Meiotic Maturation

Oxidative stress impairs the correct course of meiotic maturation, and it is known that the oocytes are exposed to increased oxidative stress during meiotic maturation in in vitro conditions. Thus, reduction of oxidative stress can lead to improved quality of cultured oocytes. The gasotransmitter carbon monoxide (CO) has a cytoprotective effect in somatic cells. The CO is produced in cells by the enzyme heme oxygenase (HO) and the heme oxygenase/carbon monoxide (HO/CO) pathway has been shown to have an antioxidant effect in somatic cells. It has not yet been investigated whether the CO has an antioxidant effect in oocytes as well. We assessed the level of expression of HO mRNA, using reverse transcription polymerase chain reaction. The HO protein localization was evaluated by the immunocytochemical method. The influence of CO or HO inhibition on meiotic maturation was evaluated in oocytes cultured in a culture medium containing CO donor (CORM-2 or CORM-A1) or HO inhibitor Zn-protoporphyrin IX (Zn-PP IX). Detection of reactive oxygen species (ROS) was performed using the oxidant-sensing probe 2′,7′-dichlorodihydrofluorescein diacetate. We demonstrated the expression of mRNA and proteins of both HO isoforms in porcine oocytes during meiotic maturation. The inhibition of HO enzymes by Zn-PP IX did not affect meiotic maturation. CO delivered by CORM-2 or CORM-A1 donors led to a reduction in the level of ROS in the oocytes during meiotic maturation. However, exogenously delivered CO also inhibited meiotic maturation, especially at higher concentrations. In summary, the CO signaling molecule has antioxidant properties in porcine oocytes and may also be involved in the regulation of meiotic maturation.

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The protective properties of synthetic porphyrin tin complexes in toxic hyperbilirubinemia

Journal of Porphyrins and Phthalocyanines ◽

10.1002/(sici)1099-1409(200004/05)4:3<243::aid-jpp201>3.0.co;2-d ◽

2000 ◽

Vol 04 (03) ◽

pp. 243-247 ◽

Cited By ~ 9

Author(s):

T. O. PHILIPPOVA ◽

B. N. GALKIN ◽

N. YA. GOLOVENKO ◽

Z. I. ZHILINA ◽

S. V. VODZINSKII

Keyword(s):

Heme Oxygenase ◽

Serum Bilirubin ◽

Protoporphyrin Ix ◽

Serum Bilirubin Level ◽

Oxygenase Activity ◽

Tetraphenyl Porphyrin ◽

Tin Complexes ◽

Cytochrome P 450

Tin complexes of meso-substituted synthetic porphyrins, namely Sn 4+-meso-tetraphenyl- porphyrin ( Sn - TPP ) and Sn 4+-meso-tetrakis(N-methyl-3-pyridyl)porphyrin tetratosylate ( Sn - TMe -3- PyP ), efficiently decrease the serum bilirubin level when injected subcutaneously at a dose of 100 μM kg−1 body weight into mice. These compounds are active during hyperbilirubinemia, induced by phenylhydrazine, hemin and tetrachloromethane, and also during autoimmune hemolytic anemia. In the latter case a decrease in serum bilirubin content was observed, as well as a decrease in the amount of blood reticulocytes which reflects a milder course of the disease. The Sn complexes under study induce, in vivo, cytochrome P-450, inhibit microsomal heme oxygenase and decrease the intensity of lipid peroxidation. At the same time, in vitro the hepatic and splenic heme oxygenase activity is blocked only when a 0.1 μM concentration of Sn - TMe -3- PyP or Sn -protoporphyrin IX is added to the incubation mixture. Sn - TPP does not affect the activity of this enzyme in vitro.

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Peer Review #2 of "Anti-apoptotic properties of carbon monoxide in porcine oocyte during in vitro aging (v0.2)"

10.7287/peerj.3876v0.2/reviews/2 ◽

2017 ◽

Keyword(s):

Carbon Monoxide ◽

Peer Review ◽

In Vitro Aging ◽

Porcine Oocyte

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Inhibitory effects of astaxanthin on postovulatory porcine oocyte aging in vitro

Scientific Reports ◽

10.1038/s41598-020-77359-6 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Bao-Yu Jia ◽

De-Cai Xiang ◽

Qing-Yong Shao ◽

Bin Zhang ◽

Shao-Na Liu ◽

...

Keyword(s):

Antioxidant Properties ◽

Underlying Mechanism ◽

Oocyte Quality ◽

Beneficial Effects ◽

Oocyte Aging ◽

Reagent Treatment ◽

Porcine Oocyte ◽

Antioxidant Gene Expression ◽

Actin Expression

AbstractMammalian oocytes represent impaired quality after undergoing a process of postovulatory aging, which can be alleviated through various effective ways such as reagent treatment. Accumulating evidences have revealed the beneficial effects of astaxanthin (Ax) as a potential antioxidant on reproductive biology. Here, porcine matured oocytes were used as a model to explore whether Ax supplement can protect against oocyte aging in vitro and the underlying mechanism, and therefore they were cultured with or without 2.5 μM Ax for an additional 24 h. Aged oocytes treated with Ax showed improved yield and quality of blastocysts as well as recovered expression of maternal genes. Importantly, oxidative stress in aged oocytes was relieved through Ax treatment, based on reduced reactive oxygen species and enhanced glutathione and antioxidant gene expression. Moreover, inhibition in apoptosis and autophagy of aged oocyte by Ax was confirmed through decreased caspase-3, cathepsin B and autophagic activities. Ax could also maintain spindle organization and actin expression, and rescue functional status of organelles including mitochondria, endoplasmic reticulum, Golgi apparatus and lysosomes according to restored fluorescence intensity. In conclusion, Ax might provide an alternative for ameliorating the oocyte quality following aging in vitro, through the mechanisms mediated by its antioxidant properties.

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173 MELATONIN ALLEVIATES THE ENDOPLASMIC RETICULUM STRESS THROUGH THE REGULATING OF UNFOLDING PROTEIN RESPONSE SIGNALING DURING PORCINE OOCYTE MATURATION IN VITRO

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab173 ◽

2017 ◽

Vol 29 (1) ◽

pp. 195 ◽

Cited By ~ 1

Author(s):

J.-Y. Park ◽

H.-J. Park ◽

J.-W. Kim ◽

S.-Y. Park ◽

S.-G. Yang ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Er Stress ◽

Oocyte Maturation ◽

Caspase 3 ◽

Oxygen Species ◽

Melatonin Treatment ◽

Maturation In Vitro ◽

Porcine Oocyte ◽

Protein Response

Unfolding protein response (UPR) is a defence mechanism during endoplasmic reticulum (ER) stress in mammalian cells. Especially, UPR genes and regulation of reactive oxygen species is involved in ER stress response on porcine oocyte maturation in vitro. Some studies have shown that melatonin treatment results in reducing oxidative stress, a protective function of free radical damage in oocyte maturation and embryo development. Also, melatonin has an important role in reducing reactive oxygen species and ER stress. However, it is unknown how the changes of UPR genes expression levels are affected the porcine oocyte maturation. In addition, there are no reports about ER stress recovery mechanism by melatonin during porcine oocyte maturation. Here, we investigated the UPR signal genes (Bip/Grp78, Atf4, p90/p50Atf6, and Xbp1) and ER-stress mediated apoptosis factors (Chop and Cleaved caspase 3) in porcine oocyte maturation in vitro. Expression of Chop and Cleaved caspase 3 mRNA levels were significantly increased (P < 0.01) in matured oocytes (metaphase II; 44 h) in vitro. Porcine oocytes were cultured in maturation medium with ER stress inducer, tunicamycin (Tm), and supplemented with various concentrations (1, 5, and 10 μg mL−1) of Tm for 0 to 44 h. Our results indicated that the proportion of matured oocytes was significantly decreased in Tm-treated groups in a dose-dependent manner (60.1 ± 1.3, 46.5 ± 2.1, and 38.9 ± 5.1% at 1, 5, and 10 μg mL−1 of Tm) compared with the control group (76.6 ± 1.9%). Likewise, mRNA expression of UPR regulator genes (Grp78/Bip, Aft4, Xbp1, Chop, and Cleaved caspase 3) was decreased by melatonin treatment (0.1 μM, 22–44 h) after pretreatment of Tm (5 μg mL−1, 0–22 h) during oocyte maturation. Our results demonstrated that the roles of melatonin as UPR signaling regulator for reducing ER stress are essential for promotion of porcine oocyte maturation and cumulus cell expansion of cumulus-oocyte complex. Moreover, the current study was initiated to confirm a functional link between effect of melatonin and regulating of UPR signaling in porcine oocytes maturation. These results suggest that melatonin improve the oocyte maturation and cumulus cells expansion by regulating of UPR signal genes against the ER stress during the porcine in vitro maturation process. This work was supported by grants from the Next-Generation BioGreen 21 Program (PJ01117604) and the Bio-industry Technology Development Program (316037–04–1-HD020) through the Rural Development Administration, the Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.

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Investigating Apoptotic Effects of Methanolic Extract of Dorema glabrum Seed on WEHI-164 Cells

ISRN Pharmacology ◽

10.1155/2013/949871 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 3

Author(s):

Maryam Bannazadeh Amirkhiz ◽

Nadereh Rashtchizadeh ◽

Hossein Nazemiyeh ◽

Jalal Abdolalizadeh ◽

Leila Mohammadnejad ◽

...

Keyword(s):

Plant Extract ◽

Caspase 3 ◽

Methanolic Extract ◽

Spherical Shape ◽

Dependent Manner ◽

Chromosomal Dna ◽

Genes Expression ◽

Apoptotic Effects ◽

Cancerous Cells

We aimed to investigate the apoptotic effects of the methanolic extract of Dorema glabrum seed on WEHI-164, cancerous cells in comparison with L929, normal cells and compared them with the cytotoxic effects of Taxol. So, MTT test and DNA fragmentation assay were performed on cultured and treated cells. Also electrophoresis which was followed by immunoblotting was done to survey the production of Caspase-3 and Bcl2 proteins, and to inquire into their relative genes expression, RT-PCR was used. According to our findings, the methanolic extract of Dorema glabrum seed can alter cells morphology as they shrink and take a spherical shape and lose their attachment too. So, the plant extract inhibits cell growth albeit in a time- and dose-dependent manner and results in degradation of chromosomal DNA. Induction of apoptosis by the plant extract was proved by the reduction of pro-Caspase-3 and Bcl2 proteins and increase in Caspase-3 gene expression and decrease in that of bcl2 too. Our data well established the antiproliferative effect of methanolic extract of Dorema glabrum seed and clearly showed that the plant extract can induce apoptosis and not necrosis in vitro. These results demonstrated that Dorema glabrum seed might be a novel and attractive therapeutic candidate for tumor treatment.

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172 Expression of proliferation and apoptosis markers in cumulus cells surrounding matured and aged oocytes exposed to luteotropic factors during the second phase of in vitro maturation

Reproduction Fertility and Development ◽

10.1071/rdv31n1ab172 ◽

2019 ◽

Vol 31 (1) ◽

pp. 210

Author(s):

I. Lebedeva ◽

O. Mityashova ◽

A. Smekalova ◽

E. Montvila ◽

G. Singina ◽

...

Keyword(s):

In Vitro Maturation ◽

Caspase 3 ◽

Cumulus Cells ◽

Treated Group ◽

Second Phase ◽

Proliferation And Apoptosis ◽

Oocyte Aging ◽

Apoptosis Markers

The quality and developmental capacity of mammalian oocytes depends on cooperation with surrounding cumulus cells. The functional state and activity of cumulus cells changes with oocyte maturation, especially during the oocyte transition from metaphase I (MI) to metaphase II (MII) stage. In the present work, effects of 3 luteotropic factors, progesterone (P4), prolactin (PRL), and LH, during the second phase of in vitro maturation (IVM) on the subsequent expression of proliferation and apoptosis markers in bovine cumulus cells surrounding matured and aged oocytes were studied. A total of 1532 cumulus-oocyte complexes (COC) were cultured for 12h in TCM-199 containing 10% fetal calf serum (FCS), 10μg mL−1 porcine FSH, and 10μg mL−1 ovine LH at 38.5°C and 5% CO2. Thereafter, COC were transferred to the following IVM systems: (1) TCM-199 containing 10% FCS (Control 1) and (2) a monolayer of granulosa cells (GC) precultured for 12h in TCM-199 containing 10% FCS (Control 2). In both systems, the medium of experimental groups was supplemented with either P4 (50 ng mL−1) or bovine PRL (50ng mL−1) or ovine LH (5μg mL−1); then, the COC were matured for next 12h. Half of the COC matured for 12h in both systems were cultured for an additional 24h in fresh TCM-199 containing 10% FCS to test long-term hormonal effects during oocyte aging. After culture, the cumulus expression of the proliferation marker proliferating cell nuclear antigen (PCNA) and the pro-apoptotic markers caspase-3 and Bax was assessed by the immunocytochemical method. The data from 4 to 5 replicates using 84 to 106 COC per treatment were analysed by ANOVA. After IVM in System 1, the rate of PCNA-positive cumulus cells was higher (P<0.05) in the PRL-treated group (41.3±1.6%) than in the control (34.6±2.3%) or LH-treated group (29.9±2.9%), but did not differ from that in the P4-treated group (38.2±4.8%). In the presence of GC (System 2), the respective rates did not change but were more variable. Aging of COC matured in both systems led to a 1.4- to 1.9-fold reduction in the proportion of the cells containing the proliferation marker PCNA (P<0.05). Meanwhile, none of the hormones tested had any long-term effect on the proliferative activity of senescent cumulus cells. The rate of cumulus cells expressing caspase-3 in different groups varied from 48.5±4.9 to 53.8±5.8% and did not depend on the hormones, IVM system, or oocyte aging. The proportion of the Bax-positive cells was also unaffected by luteotropic factors but increased 1.4 to 1.6 times (P<0.01) following 24h of COC aging. Our findings indicate that PRL can exert a short-term stimulatory action on the proliferative activity of bovine cumulus cells in the course of the second phase of IVM. Meanwhile, the cumulus expression of pro-apoptotic markers caspase-3 and Bax is not responsive to P4, PRL, or LH during the second step of IVM. The study was supported by the Russian Science Foundation (project 16-16-10069).

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Diverse Nrf2 Activators Coordinated to Cobalt Carbonyls Induce Heme Oxygenase-1 and Release Carbon Monoxide in Vitro and in Vivo

Journal of Medicinal Chemistry ◽

10.1021/acs.jmedchem.5b01509 ◽

2016 ◽

Vol 59 (2) ◽

pp. 756-762 ◽

Cited By ~ 38

Author(s):

Aniket Nikam ◽

Anthony Ollivier ◽

Michael Rivard ◽

Jayne Louise Wilson ◽

Kevin Mebarki ◽

...

Keyword(s):

Carbon Monoxide ◽

Heme Oxygenase ◽

Heme Oxygenase 1 ◽

Cobalt Carbonyls

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Carbon Monoxide Protects against Liver Failure through Nitric Oxide–induced Heme Oxygenase 1

Journal of Experimental Medicine ◽

10.1084/jem.20031003 ◽

2003 ◽

Vol 198 (11) ◽

pp. 1707-1716 ◽

Cited By ~ 157

Author(s):

Brian S. Zuckerbraun ◽

Timothy R. Billiar ◽

Sherrie L. Otterbein ◽

Peter K.M. Kim ◽

Fang Liu ◽

...

Keyword(s):

Nitric Oxide ◽

Carbon Monoxide ◽

Cell Death ◽

Protective Effect ◽

Heme Oxygenase ◽

Heme Oxygenase 1 ◽

No Production ◽

Protective Effects ◽

In Vivo Models

Carbon monoxide (CO) and nitric oxide (NO) each have mechanistically unique roles in various inflammatory disorders. Although it is known that CO can induce production of NO and that NO can induce expression of the cytoprotective enzyme heme oxygenase 1 (HO-1), there is no information whether the protective effect of CO ever requires NO production or whether either gas must induce expression of HO-1 to exert its functional effects. Using in vitro and in vivo models of tumor necrosis factor α–induced hepatocyte cell death in mice, we find that activation of nuclear factor κB and increased expression of inducible NO are required for the protective effects of CO, whereas the protective effects of NO require up-regulation of HO-1 expression. When protection from cell death is initiated by CO, NO production and HO-1 activity are each required for the protective effect showing for the first time an essential synergy between these two molecules in tandem providing potent cytoprotection.

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