scholarly journals Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos

2022 ◽  
Vol 82 ◽  
Author(s):  
P. L. Castro ◽  
A. L. J. Ferraz ◽  
J. G. Patil ◽  
R. P. Ribeiro
Keyword(s):  
Dna Fragmentation ◽  
Caspase 3 ◽  
Lower Percentage ◽  
Control Group ◽  
Oxygen Species ◽  
Apoptotic Process ◽  
Ros Formation ◽  
Apoptotic Genes ◽  
Relative Gene ◽  
Danio Rerio Embryos

Abstract This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P < 0.05) furthermore, T2 had similar expression of pro-apoptotic genes to CG (P < 0.05). ROS formation revealed that CG presented lower percentage of embryo surface area affected (3.80 ± 0.40%) (P < 0.05), in contrast, no differences were found among the other groups. T1 was most significantly (P < 0.05) damaged by DNA fragmentation. The vitrified groups with melatonin had similar damage levels of CG (P > 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.

Supplementation of kaempferol to in vitro maturation medium regulates oxidative stress and enhances subsequent embryonic development in vitro

Zygote ◽  
2019 ◽  
Vol 28 (1) ◽  
pp. 59-64
Author(s):  
Yuhan Zhao ◽  
Yongnan Xu ◽  
Yinghua Li ◽  
Qingguo Jin ◽  
Jingyu Sun ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Reactive Oxygen Species ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
Treated Group ◽  
Control Group ◽  
Oxygen Species

SummaryKaempferol (KAE) is one of the most common dietary flavonols possessing biological activities such as anticancer, anti-inflammatory and antioxidant effects. Although previous studies have reported the biological activity of KAE on a variety of cells, it is not clear whether KAE plays a similar role in oocyte and embryo in vitro culture systems. This study investigated the effect of KAE addition to in vitro maturation on the antioxidant capacity of embryos in porcine oocytes after parthenogenetic activation. The effects of kaempferol on oocyte quality in porcine oocytes were studied based on the expression of related genes, reactive oxygen species, glutathione and mitochondrial membrane potential as criteria. The rate of blastocyst formation was significantly higher in oocytes treated with 0.1 µm KAE than in control oocytes. The mRNA level of the apoptosis-related gene Caspase-3 was significantly lower in the blastocysts derived from KAE-treated oocytes than in the control group and the mRNA expression of the embryo development-related genes COX2 and SOX2 was significantly increased in the KAE-treated group compared with that in the control group. Furthermore, the level of intracellular reactive oxygen species was significantly decreased and that of glutathione was significantly increased after KAE treatment. Mitochondrial membrane potential (ΔΨm) was increased and the activity of Caspase-3 was significantly decreased in the KAE-treated group compared with that in the control group. Taken together, these results suggested that KAE is beneficial for the improvement of embryo development by inhibiting oxidative stress in porcine oocytes.

scholarly journals High expression of anti-apoptotic genes in grade I and II meningiomas

2017 ◽  
Vol 75 (4) ◽  
pp. 209-215 ◽  
Author(s):  
Daniela Pretti da Cunha Tirapelli ◽  
Sarah Bomfim Menezes ◽  
Indira Maynart Franco ◽  
Isis Lacrose Lustosa ◽  
Andressa Romualdo Rodrigues ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Control Group ◽  
Intrinsic Apoptosis ◽  
Extrinsic Pathway ◽  
Apoptosis Pathway ◽  
Intrinsic Apoptosis Pathway ◽  
Apoptosis Pathways ◽  
Genetic Mechanisms ◽  
Apoptotic Genes ◽  
Low Expression

ABSTRACT One of the different genetic mechanisms involved in the carcinogenesis of meningiomas is influenced by interactions between proteins that induce and inhibit apoptosis. Objective To evaluate the expression of c-FLIP, XIAP, Bcl-2, caspase 3, 8 and 9, cytochrome c, APAF 1 and Smac/DIABLO genes related to apoptosis pathways. Methods The gene expression was evaluated in 30 meningiomas (WHO grades I and II) and in 10 normal samples (from arachnoid tissue) through PCR-RT. Results The results showed higher expression of anti-apoptotic genes in meningiomas when compared to the control group, which had a low expression of pro-apoptotic genes. Conclusion There is a possible block in the activation of caspases through the intrinsic apoptosis pathway in meningiomas. c-FLIP modulates caspase 8 and, by inhibiting its activation due to the lack of connection with the receiver, there is a block to the FAS activation of apoptosis by its extrinsic pathway.

scholarly journals Mirabilis Jalapa L. Protein as Inductor of Apoptosis on UVB-induced Skin Damage in Mice

2017 ◽  
Vol 16 (3) ◽  
pp. 423-427
Author(s):  
Atina Hussaana ◽  
Sismindari ◽  
Sitarina Widyarini ◽  
Sudjadi ◽  
Zullies Ikawati
Keyword(s):  
Protective Effect ◽  
Dna Fragmentation ◽  
Caspase 3 ◽  
Control Group ◽  
Peak Time ◽  
Uvb Radiation ◽  
Skin Damage ◽  
Uvb Irradiation ◽  
L Protein ◽  
Mirabilis Jalapa

Background: Mirabilis jalapa L. protein (MJ-Protein) has been shown to have antioxidant and anti-inflammatory effects in vitro. Thus, it has a potential protective effect against ultraviolet B (UVB)-induced skin damage.Objective: To determine the protective effect and mechanism of MJ protein in UVB-radiation exposed mouse skin.Methods: In this experimentalstudy, 30 female BALB/c mice aged 6 weeks were exposed to a single dose of UVB irradiation with 3 minimal erythema doses (MEDs) and continued with the treatment of 0.6 mg MJ-Protein topically. The number of apoptotic body (sunburn cells) formed in epidermal layers of mouse dorsal skin was assessed at 1, 24, 48, 72, 96 and 120h after UVB irradiation was compared to that of the control group. The difference in the sunburn cells number between two groups were analyzed using independent T-test with the level of significance of 0.05. The apoptosis mechanism was confirmed qualitatively by caspase-3 and DNA fragmentation analysis in vitro.Results:At 24 h after the UVB exposure (peak time for sunburn cells formation), there was a significant increase in the sunburn cells number in the group treated with topical application of MJ-Protein. There was increased caspase-3 expression and DNA fragmentation in HeLa cells treated with MJ-Protein.Conclusions: MJ-Protein protects againts UVB-induced skin damage in mice trough apoptosis induction.Bangladesh Journal of Medical Science Vol.16(3) 2017 p.423-427

50 DEVELOPMENT OF A MODIFIED STRAW LOADING METHOD FOR VITRIFICATION OF IN VITRO-PRODUCED BOVINE BLASTOCYSTS

2014 ◽  
Vol 26 (1) ◽  
pp. 139
Author(s):  
A.-N. Ha ◽  
H.-S. Park ◽  
K.-L. Lee ◽  
Y.-G. Kim ◽  
S.-H. Song ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Cell Number ◽  
Control Group ◽  
Mrna Levels ◽  
Total Cell Number ◽  
Loading Method ◽  
Inner Surface ◽  
Apoptotic Genes ◽  
Program Grant

This study evaluated a modified plastic straw method for vitrification of in vitro-produced (IVP) bovine blastocysts. A modified straw was used that has a depressed area on its inner surface to which embryos attach. The IVP blastocysts were randomly assigned into 3 groups: (1) attachment of embryos to the inner surface of a plastic straw (aV), (2) attachment of embryos to the inner surface of a modified plastic straw (maV), and (3) non-vitrified (control). The recovery rates of blastocysts were not significantly different between the aV and maV groups (95.8 v. 94.3%; P > 0.05). The survival of post-thaw blastocysts did not significantly differ between the aV and maV groups (86.4 v. 88.2%; P > 0.05). The total cell number of blastocysts was significantly higher in the control group than in the aV and maV groups (142 ± 21.8 v. 117 ± 29.7 and 120 ± 25.2; P < 0.05), but not different between the aV and maV groups. The mRNA levels of the pro-apoptosis-related genes Bax and caspase-3 were significantly higher in the aV and maV groups than in the control group. By contrast, the mRNA levels of Bcl-2 and Mcl-1, which are anti-apoptotic genes, and of MnSOD and Prdx5, which are antioxidant-related genes, were significantly lower in the aV and maV groups than in the control group (P < 0.05). In conclusion, the aV and maV methods can be used to vitrify IVP bovine blastocysts, and embryos are more easily loaded using the maV method than using the aV method. This work was partly supported by the Next-Generation BioGreen 21 Program (grant no. PJ009587022013), IPET (grant no. 110020-5 and 112020-3), and a scholarship from the BK21 Program, Republic of Korea.

scholarly journals The Effect of D-Galactosamine on Lean and Steatotic Rat Hepatocytes in Primary Culture

2015 ◽  
pp. S637-S646 ◽  
Author(s):  
O. KUČERA ◽  
H. LOTKOVÁ ◽  
O. SOBOTKA ◽  
Z. ČERVINKOVÁ
Keyword(s):  
Lipid Peroxidation ◽  
Mitochondrial Membrane ◽  
Caspase 3 ◽  
Primary Cultures ◽  
Rat Hepatocytes ◽  
Dependent Manner ◽  
Oxygen Species ◽  
Ros Formation ◽  
Dose Dependent ◽  
Higher Sensitivity

The aim of our work was to compare the effect of D-galactosamine (GalN) on primary cultures of lean and steatotic rat hepatocytes isolated from intact and fatty liver, respectively. GalN caused more severe injury to steatotic hepatocytes than to lean cells as documented by lactate dehydrogenase leakage. Necrotic mode of cell death strongly prevails over apoptosis since we did not observe any significant increase in activities of caspase 3, 8 and 9 in any group of hepatocytes treated with GalN. Reactive oxygen species (ROS) formation and lipid peroxidation were elevated in a dose-dependent manner by GalN and were significantly more pronounced in fatty hepatocytes. A decrease in the percentage of hepatocytes with energized mitochondria was observed from 30 mM and 10 mM GalN in lean and steatotic hepatocytes, respectively. Our results undoubtedly indicate that steatotic hepatocytes exert higher sensitivity to the toxic effect of GalN. This sensitivity may be caused by more intensive GalN-induced ROS production and lipid peroxidation and by higher susceptibility of mitochondria to loss of mitochondrial membrane potential in steatotic hepatocytes. In our experimental arrangement, apoptosis does not seem to participate considerably on hepatotoxic action of GalN in either group of hepatocytes.

scholarly journals Parenteral Succinate Reduces Levels of Reactive Oxygen Species, but It Does Not Reduce Caspase-3 Serum and Tissue Levels.

2021 ◽  
Author(s):  
Sebastián Pablo Chapela ◽  
Giovanna Muscogiuri ◽  
Luigi Barrea ◽  
Evelyn Frias-Toral ◽  
Hilda Burgos ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Caspase 3 ◽  
Cecal Ligation ◽  
Krebs Cycle ◽  
Reactive Oxygen ◽  
Sepsis Group ◽  
Control Group ◽  
Oxygen Species ◽  
Sprague Dawley ◽  
Tissue Levels

Abstract Purpose: In sepsis Reactive Oxygen Species (ROS) production and apoptosis are two physiopathological processes that are increased. Succinate is a Krebs cycle intermediate that is reduced in Complex II of the mitochondria. This work aims to determine if succinate administration to septic rats improves serum ROS levels and serum and tissue levels of apoptosis. Methods: Sepsis was induced with cecal ligation and puncture in 200gr Sprague Dawley rats. 4 groups were formed (control, succinate, sepsis, and sepsis + succinate). 5mml/kg of intraperitoneal succinate were administered to the succinate and sepsis + succinate groups 2 h before sepsis induction and 2 h before sample taking. ROS levels were measured with dichlorofluorescein-diacetate, and apoptosis was measured with a Caspase-3 ELISA kit. Results: There were significant differences in serum ROS levels between the control group and the sepsis group (P = 0.012), but there were no differences in serum Caspase-3 levels (P = 0.15). The succinate administration reduced serum ROS levels in the sepsis + succinate group compared with the sepsis group in a statistically significant way (P = 0.004), but it did not reduce serum Caspase-3 levels (P = 0.39). There were no differences between groups in kidney and liver Caspase-3 levels. There was no correlation between serum ROS levels and serum Caspase-3 levels. Conclusions: In this model, ROS levels were reduced with succinate infusion, but Caspase-3 levels were not. In addition, ROS levels and apoptosis levels are not correlated, which suggests that those processes occur at different times.

Burnout of the Mind – Burnout of the Body?

2018 ◽  
Vol 32 (1) ◽  
pp. 30-42 ◽  
Author(s):  
Claudia Traunmüller ◽  
Kerstin Gaisbachgrabner ◽  
Helmut Karl Lackner ◽  
Andreas R. Schwerdtfeger
Keyword(s):  
Blood Pressure ◽  
Allostatic Load ◽  
Maslach Burnout Inventory ◽  
The Body ◽  
Lower Percentage ◽  
Control Group ◽  
Urinary Catecholamines ◽  
Morning Cortisol ◽  
The Mind ◽  
Cortisol Increase

Abstract. In the present paper we investigate whether patients with a clinical diagnosis of burnout show physiological signs of burden across multiple physiological systems referred to as allostatic load (AL). Measures of the sympathetic-adrenergic-medullary (SAM) axis and the hypothalamic-pituitary-adrenal (HPA) axis were assessed. We examined patients who had been diagnosed with burnout by their physicians (n = 32) and were also identified as burnout patients based on their score in the Maslach Burnout Inventory-General Survey (MBI-GS) and compared them with a nonclinical control group (n = 19) with regard to indicators of allostatic load (i.e., ambulatory ECG, nocturnal urinary catecholamines, salivary morning cortisol secretion, blood pressure, and waist-to-hip ratio [WHR]). Contrary to expectations, a higher AL index suggesting elevated load in several of the parameters of the HPA and SAM axes was found in the control group but not in the burnout group. The control group showed higher norepinephrine values, higher blood pressure, higher WHR, higher sympathovagal balance, and lower percentage of cortisol increase within the first hour after awakening as compared to the patient group. Burnout was not associated with AL. Results seem to indicate a discrepancy between self-reported burnout symptoms and psychobiological load.

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