Eicosapentaenoic acid supplemented to in vitro maturation medium results in lesser lipid content and intracellular reactive oxygen species in blastocysts of cattle

2021 ◽  
pp. 106765
Author(s):  
Noelia Nikoloff ◽  
Ana C. Carranza ◽  
Mariana C. Fabra ◽  
Anabella Campagna ◽  
Juan P. Anchordoquy ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Eicosapentaenoic Acid ◽  
Lipid Content ◽  
In Vitro Maturation ◽  
Reactive Oxygen ◽  
Intracellular Reactive Oxygen Species ◽  
Oxygen Species ◽  
Maturation Medium

Reduced levels of intracellular reactive oxygen species and apoptotic status are not correlated with increases in cryotolerance of bovine embryos produced in vitro in the presence of antioxidants

2014 ◽  
Vol 26 (6) ◽  
pp. 797 ◽  
Author(s):  
Nathália A. S. Rocha-Frigoni ◽  
Beatriz C. S. Leão ◽  
Ériklis Nogueira ◽  
Mônica F. Accorsi ◽  
Gisele Z. Mingoti
Keyword(s):  
Reactive Oxygen Species ◽  
Reactive Oxygen ◽  
Intracellular Reactive Oxygen Species ◽  
Control Group ◽  
Bovine Embryo ◽  
Oxygen Species ◽  
Antioxidant Supplementation ◽  
In Vitro Production ◽  
Intracellular Ros

The effects of intracellular (cysteine and β-mercaptoethanol) and extracellular (catalase) antioxidant supplementation at different times during in vitro production (IVM and/or in vitro culture (IVC)) on bovine embryo development, intracellular reactive oxygen species (ROS) levels, apoptosis and re-expansion rates after a vitrification–thawing process were examined. Blastocyst frequencies were not affected by either antioxidant supplementation (40.5%–56.4%) or the timing of supplementation (41.7%–55.4%) compared with control (48.7%; P > 0.05). Similarly, antioxidants and the moment of supplementation did not affect (P > 0.05) the total number of blastomeres (86.2–90.5 and 84.4–90.5, respectively) compared with control (85.7). However, the percentage of apoptotic cells was reduced (P < 0.05) in groups supplemented during IVM (1.7%), IVC (2.0%) or both (1.8%) compared with control (4.3%). Intracellular ROS levels measured in Day 7 blastocysts were reduced (P < 0.05) in all groups (0.60–0.78), with the exception of the group supplemented with β-mercaptoethanol during IVC (0.88), which did not differ (P > 0.05) from that in the control group (1.00). Re-expansion rates were not affected (P > 0.05) by the treatments (50.0%–93.0%). In conclusion, antioxidant supplementation during IVM and/or IVC reduces intracellular ROS and the rate of apoptosis; however, supplementation does not increase embryonic development and survival after vitrification.

scholarly journals Protective Effect of a Fucose-Rich Fucoidan Isolated from Saccharina japonica against Ultraviolet B-Induced Photodamage In Vitro in Human Keratinocytes and In Vivo in Zebrafish

Marine Drugs ◽  
2020 ◽  
Vol 18 (6) ◽  
pp. 316 ◽  
Author(s):  
Wanchun Su ◽  
Lei Wang ◽  
Xiaoting Fu ◽  
Liying Ni ◽  
Delin Duan ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Protective Effect ◽  
Reactive Oxygen ◽  
Saccharina Japonica ◽  
Intracellular Reactive Oxygen Species ◽  
Dependent Manner ◽  
Hacat Cells ◽  
Oxygen Species

A fucose-rich fucoidan was purified from brown seaweed Saccharina japonica, of which the UVB protective effect was investigated in vitro in keratinocytes of HaCaT cells and in vivo in zebrafish. The intracellular reactive oxygen species levels and the viability of UVB-irradiated HaCaT cells were determined. The results indicate that the purified fucoidan significantly reduced the intracellular reactive oxygen species levels and improved the viability of UVB-irradiated HaCaT cells. Furthermore, the purified fucoidan remarkably decreased the apoptosis by regulating the expressions of Bax/Bcl-xL and cleaved caspase-3 in UVB-irradiated HaCaT cells in a dose-dependent manner. In addition, the in vivo UV protective effect of the purified fucoidan was investigated using a zebrafish model. It significantly reduced the intracellular reactive oxygen species level, the cell death, the NO production, and the lipid peroxidation in UVB-irradiated zebrafish in a dose-dependent manner. These results suggest that purified fucoidan has a great potential to be developed as a natural anti-UVB agent applied in the cosmetic industry.

Effects of Reactive Oxygen Species and Antioxidants during In vitro Maturation Oocytes and Embryo Development in Pigs

2017 ◽  
Vol 41 (1) ◽  
pp. 17-23
Author(s):  
Won-Hee Lee ◽  
◽  
Ji-Eun Park ◽  
Yong Hwangbo ◽  
Hwa-Young Kim ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Embryo Development ◽  
In Vitro Maturation ◽  
Reactive Oxygen ◽  
Oxygen Species

scholarly journals Hyperthermia Sensitizes Rhizopus oryzae to Posaconazole and Itraconazole Action through Apoptosis

2013 ◽  
Vol 57 (9) ◽  
pp. 4360-4368 ◽  
Author(s):  
Fazal Shirazi ◽  
Michael A. Pontikos ◽  
Thomas J. Walsh ◽  
Nathaniel Albert ◽  
Russell E. Lewis ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Rhizopus Oryzae ◽  
Reactive Oxygen ◽  
Annexin V ◽  
Intracellular Reactive Oxygen Species ◽  
Oxygen Species ◽  
Content Type ◽  
Dependent Inhibition ◽  
Dose Dependent Inhibition

ABSTRACTThe high mortality rate of mucormycosis with currently available monotherapy has created interest in studying novel strategies for antifungal agents. With the exception of amphotericin B (AMB), the triazoles (posaconazole [PCZ] and itraconazole [ICZ]) are fungistaticin vitroagainstRhizopus oryzae. We hypothesized that growth at a high temperature (42°C) results in fungicidal activity of PCZ and ICZ that is mediated through apoptosis.R. oryzaehad high MIC values for PCZ and ICZ (16 to 64 μg/ml) at 25°C; in contrast, the MICs for PCZ and ICZ were significantly lower at 37°C (8 to 16 μg/ml) and 42°C (0.25 to 1 μg/ml). Furthermore, PCZ and ICZ dose-dependent inhibition of germination was more pronounced at 42°C than at 37°C. In addition, intracellular reactive oxygen species (ROS) increased significantly when fungi were exposed to antifungals at 42°C. Characteristic cellular changes of apoptosis inR. oryzaewere induced by the accumulation of intracellular reactive oxygen species. Cells treated with PCZ or ICZ in combination with hyperthermia (42°C) exhibited characteristic markers of early apoptosis: phosphatidylserine externalization visualized by annexin V staining, membrane depolarization visualized by bis-[1,3-dibutylbarbituric acid] trimethine oxonol (DiBAC) staining, and increased metacaspase activity. Moreover, terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and DAPI (4′,6-diamidino-2-phenylindole) staining demonstrated DNA fragmentation and condensation, respectively. The addition ofN-acetylcysteine increased fungal survival, prevented apoptosis, reduced ROS accumulation, and decreased metacaspase activation. We concluded that hyperthermia, either alone or in the presence of PCZ or ICZ, induces apoptosis inR. oryzae. Local thermal delivery could be a therapeutically useful adjunct strategy for these refractory infections.

203 SUPPLEMENTATION WITH LINOLENIC ACID, L-CARNITINE, ALONE OR ASSOCIATED, DURING IVM RESULTED IN DECREASE OF ROS LEVELS AND APOPTOTIC INDEX OF BOVINE IN VITRO PRODUCED EMBRYOS

2016 ◽  
Vol 28 (2) ◽  
pp. 232
Author(s):  
B. C. S. Leão ◽  
N. A. S. R. Frigoni ◽  
P. C. Dall'Acqua ◽  
M. Ambrogi ◽  
G. Z. Mingoti
Keyword(s):  
Reactive Oxygen Species ◽  
Embryonic Development ◽  
Linolenic Acid ◽  
Lipid Content ◽  
Reactive Oxygen ◽  
Antioxidant Effect ◽  
Apoptotic Index ◽  
Control Group ◽  
Oxygen Species

Supplementation of in vitro maturation (IVM) medium with linolenic acid (ALA) has been used in order to reduce oocyte lipid content and have beneficial effects on maturation and acquisition of competence for embryonic development. Besides the effect of reducing cellular lipid content, l-carnitine (l-car) has an antioxidant effect by reducing the levels of reactive oxygen species (ROS) and protecting cells from apoptosis. However, the association of ALA and l-car has never been tested. This study was conducted to evaluate the effects of supplementation of IVM medium of bovine oocytes with ALA, l-car or the association of both (ALA+l-car) on embryonic development and blastocysts reactive oxygen species (ROS) levels and occurrence of apoptosis. Cumulus-oocyte complexes (n = 2241, in 11 replicates) were matured during 22 h at 38.5°C and 5% CO2 in air, in TCM-199 medium with bicarbonate, hormones and 10% FCS (control group), also supplemented with 100 μM ALA group; or 5 mM l-car (l-car group); or 100 μM ALA associated with 5 mM l-car (ALA+l-car group). After fertilisation (Day 0), zygotes were cultured 7 days in SOF that was supplemented with 0.5% BSA and 2.5% FCS, in 5% CO2 in air at 38.5°C. The cleavage and blastocysts rates were evaluated, respectively, at Days 3 and 7. Blastocysts were stained with 5 mM of H2DCFDA (Molecular Probes, Invitrogen, Carlsbad, CA, USA) and TUNEL (In Situ Cell Death Detection Kit, Roche Applied Science, Boston, MA, USA), to evaluate the ROS levels and the blastomers apoptotic index, respectively. The ROS (n = 115) and TUNEL (n = 102) stained blastocysts were evaluated under an epifluorescence microscope (excitation 495 nm/510–550 nm and emission 404 nm/590 nm), and the ROS levels (expressed as arbitrary fluorescence units) were measured by Q-Capture Pro image software (Q Imaging, Surrey, BC, Canada). The fluorescence intensity values were subtracted from mean values of background in the images. The variables were analysed by ANOVA followed by Tukey’s test (P < 0.05) and data are presented as mean ± s.e.m. There was no effect (P > 0.05) of the supplements during IVM on cleavage and blastocysts rates (%), respectively, for control (81.1 ± 1.8 and 29.0 ± 3.1), ALA (80.5 ± 2.1 and 29.7 ± 2.3), l-car (79.5 ± 2.8 and 29.2 ± 2.3), and ALA+l-car (82.2 ± 1.1 and 30.5 ± 2.0) groups. The oocytes supplementation resulted in a decrease (P < 0.05) in ROS levels for ALA (0.84 ± 0.04), l-car (0.85 ± 0.03) and ALA+l-car (0.82 ± 0.02) groups, compared to the Control (1.00 ± 0.05). Consequently, the percentage of apoptotic blastomeres decreased (P < 0.05) after ALA (6.9 ± 1.0%), l-car (7.5 ± 1.2%) and ALA+l-car (4.6 ± 0.7%) supplementations, unlike to the Control group (12.0 ± 1.2%). In conclusion, the supplementation with ALA, l-car or ALA+l-car during IVM did not affect the blastocyst development, but led to a reduction in ROS levels and in the apoptotic index of such blastocysts. These findings may be due to some antioxidant effect of these supplements in the oocytes and/or the produced embryos. Financial support was through FAPESP (#2012/10084–4 and #2013/07382–6).

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