184 Effects of cyanidin supplementation on invitro maturation of pig oocytes

2020 ◽  
Vol 32 (2) ◽  
pp. 220
Author(s):  
E. Hicks ◽  
M. Mentler ◽  
B. D. Whitaker

Oxidative stress can have a negative effect on oocyte maturation during invitro production of pig embryos. Imbalance of reactive oxygen species and antioxidant levels can affect the progression of oocyte maturation up to the point of fertilization. Antioxidants are effective in maintaining more ideal reactive oxygen species levels, which help to protect oocytes from potential harmful effects of oxidative stress. Berries from the elder plant (Sambucus sp.) contain high levels of a broad spectrum of antioxidants. One of these antioxidants, cyanidin, when supplemented to maturation medium at 100μM concentrations, reduces reactive oxygen species formation and improves IVF and early embryonic development in pigs. However, changes in the enzyme mechanisms of action during oocyte maturation due to cyanidin supplementation are unknown. Therefore, the objective of this study was to characterise the intracellular oocyte enzyme mechanisms between oocytes supplemented with 100μM cyanidin during 40 to 44h of maturation (n=600) and oocytes without supplementation of cyanidin during maturation (n=558). At the end of maturation, oocytes were evaluated for either glutathione peroxidase (n=300), catalase (n=564), or superoxide dismutase (n=294) activities. Glutathione peroxidase activity was determined by following the rate of NADPH oxidation, catalase activity was determined by following the rate of hydrogen peroxide decomposition, and superoxide dismutase activity was determined by following the reduction rate of cytochrome c, utilising the xanthine-xanthine oxidase system. Data were analysed using ANOVA and Tukey's test. There were no significant differences between oocytes matured with 100μM cyanidin and those that were not when comparing glutathione peroxidase and superoxide dismutase activities. Supplementation of 100μM cyanidin to maturation medium increased (P<0.05) catalase activity in oocytes (0.78±0.15 units/oocyte) compared with no cyanidin supplementation (0.14±0.11 units/oocyte). These results indicate that supplementing 100μM cyanidin to the maturation medium of pig oocytes could reduce the negative effects of oxidative stress by increasing intracellular catalase activity during oocyte maturation.

2017 ◽  
Vol 43 (2) ◽  
pp. 626-635 ◽  
Author(s):  
Yan-Yan Zhang ◽  
Minhan Yi ◽  
Yong-Pan Huang

Background/Aims: Doxorubicin-induced cardiac toxicity has been a major concern of oncologists and is considered the main restriction on its clinical application. Oxymatrine has shown potent anti-cancer, anti-fibrosis, and anti-oxidative effects. Recently, it has been reported that oxymatrine is protective against some cardiovascular diseases. In this study, we aimed to investigate the effects of oxymatrine on doxorubicin-induced cardiotoxicity in rat hearts and H9c2 cells. Methods: Creatine Kinase - MB (CK-MB) and Lactate Dehydrogenase (LDH) levels were determined using commercial kits. Biochemical indices reflecting oxidative stress, such as catalase (CAT), malonyldialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were also analyzed with commercial kits. Mitochondrial reactive oxygen species (ROS) 2’,7’-dichlorofluorescin diacetate (DCFH-DA) was measured by fluorescence microscopy. Histological analyses were conducted to observe morphological changes, and apoptosis was measured using a commercial kit. Western blots were used to detect the level of expression of cleaved caspase-3. Results: Doxorubicin treatment significantly increased oxidative stress levels, as indicated by catalase, malonyldialdehyde, superoxide dismutase, glutathione peroxidase and reactive oxygen species. Doxorubicin also increased pathological damage in myocardial tissue, myocardial ROS levels, and malonyldialdehyde levels, and induced apoptosis in myocardial tissues and H9c2 cells. All of these doxorubicin-induced effects were attenuated by oxymatrine. Conclusion: These in vitro and in vivo findings indicate that oxymatrine may be a promising cardioprotective agent against doxorubicin-induced cardiotoxicity, at least in part mediated through oxymatrine’s inhibition of cardiac apoptosis and oxidative stress.


Author(s):  
А.Г. Кутихин ◽  
Д.К. Шишкова ◽  
Р.А. Мухамадияров ◽  
Е.А. Великанова

Введение. Кальций-фосфатные бионы (КФБ) формируются в организме человека при перенасыщении сыворотки ионами кальция и фосфора и вызывают дисфункцию эндотелия, однако молекулярные механизмы нарушения функционирования эндотелия при воздействии КФБ не ясны. Цель исследования - выяснение роли кальций-фосфатных бионов различной формы в развитии окислительного стресса в артериальных эндотелиальных клетках (ЭК) человека. Методика. Для детекции окислительного стресса к конфлюэнтным культурам первичных ЭК коронарной и внутренней грудной артерии человека добавляли равные концентрации КФБ сферической или игольчатой формы (СКФБ и ИКФБ соответственно) с последующим культивированием в течение 1 и 4 ч, добавлением флюоресцентных индикаторов окислительного стресса MitoSOX Red и CellROX Green и конфокальной микроскопией. Измеряли концентрацию продуктов перекисного окисления липидов в культуральной жидкости через 24 ч экспозиции эндотелиальных клеток КФБ. Анализ нейтрализации цитотоксических эффектов перекисного окисления липидов проводили путем добавления к ЭК супероксиддисмутазы и каталазы на 4 или 24 ч (одновременно с КФБ). Для сравнения механизмов клеточной гибели при воздействии СКФБ и ИКФБ анализировали цитотоксичность обоих типов бионов при одновременном воздействии лизосомального ингибитора бафиломицина А1. Результаты. Значимого увеличения генерации активных форм кислорода (АФК) в результате экспозиции СКФБ (независимо от линии ЭК и продолжительности экспозиции) не было выявлено. В то же время наблюдалось повышение генерации супероксида через 4 ч, а иных свободных радикалов через 1 ч после добавления ИКФБ к ЭК. Предварительная нейтрализация АФК супероксиддисмутазой и каталазой частично защищала ЭК от индуцируемой ИКФБ гибели. При этом добавление бафиломицина А1 к ЭК частично защищало их от гибели только при воздействии СКФБ, но не ИКФБ. Заключение. Гибель ЭК при воздействии СКФБ происходит в результате первичного повреждения лизосом, а при воздействии ИКФБ - в первую очередь вследствие окислительного стресса. Background. Calcium phosphate bions (CPB) form in the human blood upon its supersaturation with calcium and phosphate and provoke endothelial dysfunction; however, the molecular mechanisms of these pathological processes remain unclear. Aim. To elucidate the role of differently shaped CPBs in induction of oxidative stress in human arterial endothelial cells (Ecs). Methods. For detection of oxidative stress, equal concentrations of spherical CPB (CPB-S) or needle-shaped CPB (CPB-N) were added to confluent cultures of primary human coronary artery and internal thoracic artery ECs for 1 and 4 h; this was followed by MitoSOX Red and CellROX Green staining and subsequent confocal microscopy. Concentration of thiobarbituric acid-reactive substances was measured in the EC culture supernatant at 24 h of the CPB exposure. The lipid peroxidation cytotoxicity was neutralized by adding superoxide dismutase and catalase to ECs for 4 or 24 h. To compare cell death subroutines induced by CPB-S and CPB-N, the effect of bafilomycin A1, a lysosomal inhibitor, on CRB cytotoxicity was studied. Results. No increase in reactive oxygen species generation was observed in the CPB-S exposure, regardless of the EC line and exposure duration. However, addition of CPB-N to ECs increased the production of superoxide and other free radicals after four- and one-hour exposure, respectively. Prior neutralization of reactive oxygen species with superoxide dismutase and catalase partially protected ECs from CPB-N- but not CPB-S-induced death while bafilomycin A1, vice versa, protected ECs from CPB-S- but not CPB-N-induced death. Conclusion. CPB-S cause cell death due to primary damage of lysosomes whereas CPB-N induce apoptosis due to oxidative stress.


Author(s):  
Mojdeh Hosseinpoor Kashani ◽  
Mina Ramezani ◽  
Zeinab Piravar

Background: Acrylamide (AA) is a reactive molecule produced during food processing at temperatures above 120°C. Objective: To evaluate the impact of different concentrations of AA on human sperm parameters, oxidative stress and total antioxidant capacity (TAC). Materials and Methods: In this laboratory study, semen samples were obtained from healthy donors referred to the Taleghani Hospital, Tehran, Iran between June and July 2019. Samples were divided into four groups (n = 10/each): one control and three treatment groups (0.5, 1, and 2 mM of AA). After 2 hr of exposure to AA, the superoxide dismutase and malondialdehyde levels were measured based on colorimetric methods. The TAC was determined by the ferric-reducing antioxidant power assay. Flow cytometry was performed to measure the intracellular reactive oxygen species generation. Also, immunohistochemistry was done to determine the effect of AA on tyrosine phosphorylation and carboxymethyl-lysine expression. Results: Results of the study demonstrated that the motility and viability of spermatozoa were significantly decreased after AA exposure (p < 0.001). This decrease was also seen in the TAC and superoxide dismutase activity as well as in the phosphotyrosine percentage compared with the control (p < 0.01). However, the carboxymethyllysine and prooxidant activity including reactive oxygen species generation and lipid peroxidation level increased (p < 0.001). Conclusion: Overall, the results confirmed the detrimental effect of AA on human spermatozoa which may be due to oxidative stress and decreased total antioxidant levels. AA may reduce fertility by reducing sperm capacitation and motility. Key words: Acrylamide, Oxidative stress, Antioxidant, Spermatozoa, Infertility.


2019 ◽  
Vol 31 (1) ◽  
pp. 209
Author(s):  
M. Mentler ◽  
J. Current ◽  
B. Whitaker

Elevated environmental temperatures induce heat stress, which can cause a depression in fertility and early embryonic development. Fatty acids initiate an endergonic reaction that is able to absorb cellular heat, causing a decrease in intracellular temperature. Supplementing linoleic and linolenic acids to the maturation medium of pig oocytes at elevated temperatures reduces the effects of heat stress-induced damage during fertilization and embryonic development. However, the mechanism of action of fatty acids during oocyte maturation is unknown. Therefore, the objective of this study was to minimize heat stress-induced damage and characterise the intracellular oocyte mechanisms. Oocyte maturation media was supplemented with linoleic and linolenic acid during oocyte maturation at either 38.5 or 41.5°C. Oocytes (n=3094, r=5) were supplemented with 50μM linoleic acid, 50μM linolenic acid, 25μM of both, or 50μM of both during 40 to 44h of maturation and then evaluated for the formation of reactive oxygen species (n=239), intracellular glutathione concentrations (n=1005), glutathione peroxidase (n=1005), catalase (n=987), and superoxide dismutase (n=863) activities. Data were analysed using ANOVA with the main effects including treatment, well, and replicate. There were no significant differences between the treatment groups matured at 38.5°C when comparing reactive oxygen species generation. Supplementation of linoleic or linolenic acid significantly decreased (P&lt;0.05) reactive oxygen species generation in oocytes matured at 41.5°C compared with no supplementation at the same temperature. Supplementation of linoleic or linolenic acid or both significantly increased (P&lt;0.05) intracellular glutathione concentrations compared with no supplementation at 38.5°C (23.37±1.23 pmol/oocyte) and 41.5°C (10.42±1.01 pmol/oocyte). There were no significant differences between the treatment groups matured at 38.5°C or 41.5°C when comparing glutathione peroxidase activity. Supplementation of linoleic or linolenic acid or both significantly increased (P&lt;0.05) catalase and superoxide dismutase activities compared with no supplementation at 38.5°C and at 41.5°C. Superoxide dismutase activity was significantly higher (P&lt;0.05) in oocytes matured at 41.5°C compared with those matured at 38.5°C. These results indicate that supplementing linoleic and linolenic acid to the maturation medium of pig oocytes at an elevated temperature reduces the effects of heat stress-induced damage by increasing intracellular glutathione concentrations and catalase and superoxide dismutase activities.


2018 ◽  
Vol 47 (2) ◽  
pp. 531-537
Author(s):  
Damaris OJEDA-BARRIOS ◽  
Jorge CASTILLO-GONZALEZ ◽  
Adriana HERNANDEZ-RODRIGUEZ ◽  
Javier ABADIA ◽  
Estaban SANCHEZ ◽  
...  

In the United States of America and in Mexico, zinc deficiency is a common nutritional disorder in pecan trees [Carya illinoinensis (Wangenh.) C. Koch], especially in calcareous soils. This study in Chihuahua, northern Mexico, analyses the effects of zinc nutritional status on various physiological and nutritional indicators, on the metabolism of oxidative stress, and on the yield and fruit quality of pecan. The aim was to identify possible bioindicators of soil zinc deficiency. The experimental design was completely randomized with four nutritional conditions with respect to zinc: a control and three levels of zinc deficiency - slight, moderate and severe. Zinc deficiency is characterised by small leaves with interveinal necrosis and rippled leaf margins. The lowest values of leaf area, SPAD values, total N and NO3 concentration were observed under conditions of severe zinc deficiency. With worsening zinc deficiency, results indicate an increased enzymatic activity of superoxide dismutase, catalase and glutathione peroxidase. Interestingly, under severe zinc deficiency there are decreases in trunk cross-sectional area growth, in yield and in percentage kernel. Increased activity of superoxide dismutase, catalase and peroxidase enzymes is associated with detoxification of reactive oxygen species. The activity of enzymes detoxifying reactive oxygen species lessens the negative effects of zinc deficiency stress, and may be good bioindicators of zinc deficiency and its visual symptoms on pecan trees.


2006 ◽  
Vol 291 (5) ◽  
pp. H2522-H2532 ◽  
Author(s):  
Kyoko Umeji ◽  
Seiji Umemoto ◽  
Shinichi Itoh ◽  
Masakazu Tanaka ◽  
Shinji Kawahara ◽  
...  

Reactive oxygen species-scavenging enzyme Cu/Zn superoxide dismutase (SOD) regulated by peroxisome proliferator-activated receptors (PPARs) plays an important role in vascular responsiveness. However, it remains unknown whether statin restores vascular dysfunction through the activation of reactive oxygen species-scavenging enzymes in vivo. We hypothesized that pitavastatin restores vascular function by modulating oxidative stress through the activation of Cu/ZnSOD and PPAR-γ in hypercholesterolemia. New Zealand White male rabbits were fed either normal chow or a 1% cholesterol (CHO) diet for 14 wk. After the first 7 wk, the CHO-fed rabbits were further divided into three groups: those fed with CHO feed only (HC), those additionally given pitavastatin, and those additionally given an antioxidant, probucol. The extent of atherosclerosis was assessed by examining aortic stiffness. When compared with the HC group, both the pitavastatin and probucol groups showed improved aortic stiffness by reducing aortic levels of reactive oxidative stress, nitrotyrosine, and collagen, without affecting serum cholesterol or blood pressure levels. Pitavastatin restored both Cu/ZnSOD activity ( P < 0.005) and PPAR-γ expression and activity ( P < 0.01) and inhibited NAD(P)H oxidase activity ( P < 0.0001) in the aorta, whereas probucol inhibited NAD(P)H oxidase activity more than did pitavastatin ( P < 0.0005) without affecting Cu/ZnSOD activity or PPAR-γ expression and activity. Importantly, Cu/ZnSOD activity was positively correlated with the PPAR-γ activity in the aorta ( P < 0.005), both of which were negatively correlated with aortic stiffness ( P < 0.05). Vascular Cu/ZnSOD and PPAR-γ may play a crucial role in the antiatherogenic effects of pitavastatin in hypercholesterolemia in vivo.


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