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1.
Toxicol Appl Pharmacol ; 485: 116910, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38521372

RESUMO

3-nitropropionic acid (3-NPA), a toxic metabolite produced by mold, is mainly found in moldy sugarcane. 3-NPA inhibits the activity of succinate dehydrogenase that can induce oxidative stress injury in cells, reduce ATP production and induce oxidative stress in mouse ovaries to cause reproductive disorders. Ursolic acid (UA) has a variety of biological activities and is a pentacyclic triterpene compound found in many plants. This experiment aimed to investigate the cytotoxicity of 3-NPA during mouse oocyte in vitro maturation and the protective effects of UA on oocytes challenged with 3-NPA. The results showed that UA could alleviate 3-NPA-induced oocyte meiotic maturation failure. Specifically, 3-NPA induced a decrease in the first polar body extrusion rate of oocytes, abnormal distribution of cortical granules, and an increase in the proportion of spindle abnormalities. In addition, 3-NPA caused mitochondrial dysfunction and induced oxidative stress, including decreases in the GSH, mitochondrial membrane potential and ATP levels, and increases in the ROS levels, and these effects led to apoptosis and autophagy. The addition of UA could significantly improve the adverse effects caused by 3-NPA. In general, our data show that 3-NPA affects the normal development of oocytes during the in vitro culture, and the addition of UA can effectively repair the damage caused by 3-NPA to oocytes.


Assuntos
Meiose , Nitrocompostos , Oócitos , Estresse Oxidativo , Propionatos , Triterpenos , Ácido Ursólico , Animais , Nitrocompostos/toxicidade , Propionatos/toxicidade , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Feminino , Meiose/efeitos dos fármacos , Camundongos , Triterpenos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Apoptose/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Autofagia/efeitos dos fármacos , Trifosfato de Adenosina/metabolismo , Camundongos Endogâmicos ICR
2.
Zygote ; 30(5): 704-711, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35677960

RESUMO

Glyphosate is a universal herbicide with genital toxicity, but the effect of glyphosate on oocytes has not been reported. This study aimed to evaluate the effect of glyphosate (0, 10, 20, 50 and 100 mM) on bovine oocyte in vitro maturation. We showed that 50 mM glyphosate adversely affects the development of bovine oocytes. Exposure of oocytes to 50 mM glyphosate caused an abnormal reduction in oxidative (redox) levels compared with that in the control group, with a significantly higher reactive oxide species level (P < 0.05) and significantly lower glutathione (GSH) expression (P < 0.05). Additionally, the mRNA levels of antioxidant genes (SOD1, SOD2, SIRT2, SIRT3) and the mitochondrial membrane potential (MMP) were significantly reduced (P < 0.05). Furthermore, treatment with 50 mM glyphosate-induced apoptosis, and the mRNA levels of the apoptotic genes Caspase-3 and Caspase-4 were significantly higher than those in the control group (P < 0.05); however, the mRNA level of BAX was significantly higher than that in the control group (P < 0.01). Additionally, the mRNA levels of the anti-apoptotic genes Survivin and BCL-XL were significantly lower than those in the control group (P < 0.05), and oocyte quality was adversely affected. Together, our results confirmed that glyphosate impairs the quality of oocytes by promoting abnormal oocyte redox levels and apoptosis.


Assuntos
Herbicidas , Sirtuína 3 , Animais , Antioxidantes/metabolismo , Apoptose , Caspase 3/genética , Caspase 3/metabolismo , Bovinos , Glutationa/metabolismo , Glicina/análogos & derivados , Herbicidas/metabolismo , Herbicidas/toxicidade , Técnicas de Maturação in Vitro de Oócitos/métodos , Oócitos , Estresse Oxidativo , Óxidos/metabolismo , Óxidos/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sirtuína 2/metabolismo , Sirtuína 2/farmacologia , Sirtuína 3/metabolismo , Sirtuína 3/farmacologia , Superóxido Dismutase-1 , Survivina/metabolismo , Survivina/farmacologia , Proteína X Associada a bcl-2 , Glifosato
3.
Zygote ; 30(1): 57-64, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33966682

RESUMO

Methomyl is a widely used carbamate insecticide and environmental oestrogen that has adverse effects on the reproductive system. However, there have been no reports on the effect of methomyl on early embryos in mammals. In this study, we explored the effect of methomyl exposure on the quality of early embryonic development in mice and the possible mechanisms. During in vitro culture, different concentrations of methomyl (10, 20, 30 and 35 µM) were added to mouse zygote medium. The results showed that methomyl had an adverse effect on early embryonic development. Compared with the control group, the addition of 30 µM methomyl significantly reduced the rate of early embryo blastocyst formation. Methomyl exposure can increase oxidative stress and impair mitochondrial function, which may be the cause of blastocyst formation. In addition, we found that methomyl exposure promoted apoptosis and autophagy in mouse blastocysts. The toxic effect of methomyl on early embryos may be the result of oxidative stress induction. Taken together, our results indicate that methomyl can cause embryonic development defects in mice, thereby reducing the quality of early embryo development.


Assuntos
Desenvolvimento Embrionário , Metomil , Animais , Blastocisto/metabolismo , Técnicas de Cultura Embrionária , Embrião de Mamíferos , Feminino , Metomil/metabolismo , Camundongos , Estresse Oxidativo , Gravidez
4.
Zygote ; 30(3): 358-364, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34676817

RESUMO

Methomyl is a broad-spectrum carbamate insecticide that has a variety of toxic effects on humans and animals. However, there have been no studies on the toxicity of methomyl in female mammalian oocytes. This study investigated the toxic effects of environmental oestrogen methomyl exposure on mouse oocyte maturation and its possible mechanisms. Our results indicated that methomyl exposure inhibited polar body extrusion in mouse oocytes. Compared with that in the control group, in the methomyl treatment group, superoxide anion free radicals in oocytes were significantly increased. In addition, the mitochondrial membrane potential of metaphase II stage oocytes in the methomyl treatment group was significantly decreased, resulting in reduced mouse oocyte quality. After 8.5 h of exposure to methomyl, metaphase I stage mouse oocytes displayed an abnormal spindle morphology. mRNA expression of the pro-apoptotic genes Bax and Caspase-3 in methomyl-treated oocytes increased, which confirmed the apoptosis. Collectively, our results indicated that mouse oocyte maturation is defective after methomyl treatment at least through disruption of spindle morphology, mitochondrial function and by induction of oxidative stress.


Assuntos
Metomil , Oócitos , Animais , Feminino , Mamíferos , Metáfase , Metomil/metabolismo , Metomil/farmacologia , Camundongos , Mitocôndrias , Oogênese
5.
Zygote ; 28(1): 59-64, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31662136

RESUMO

Kaempferol (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.


Assuntos
Embrião de Mamíferos/citologia , Desenvolvimento Embrionário/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Maturação in Vitro de Oócitos , Quempferóis/farmacologia , Oócitos/citologia , Estresse Oxidativo/efeitos dos fármacos , Animais , Embrião de Mamíferos/efeitos dos fármacos , Embrião de Mamíferos/metabolismo , Feminino , Glutationa/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Suínos
6.
J Reprod Dev ; 65(6): 499-506, 2019 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-31474647

RESUMO

L-carnitine (LC) is well known for its antioxidant activity. In this study, we explored the potential mechanistic effects of LC supplementation on aged bovine oocytes in vitro. We showed that in-vitro maturation could enhance the subsequent developmental capacity of aging oocytes, when supplemented with LC. After in vitro fertilization, the blastocyst formation rate in the aged oocytes post-LC treatment significantly increased compared to that in untreated aged oocytes (29.23 ± 2.20% vs. 20.90 ± 3.05%). Furthermore, after LC treatment, the level of intracellular reactive oxygen species in aged oocytes significantly decreased, and glutathione levels significantly increased, compared to those in untreated aged oocytes. Mitochondrial membrane potential, the percentage of early apoptotic oocytes, and caspase-3 activity were significantly reduced in LC-treated aged oocytes compared to those in untreated aged oocytes. Furthermore, during in vitro aging, the mRNA levels of the anti-apoptotic genes, Bcl-xl and survivin in LC-treated aged oocytes were significantly higher than those in untreated aged oocytes. Overall, these results indicate that at least in in vitro conditions, LC can prevent the aging of bovine oocytes and improve the developmental capacity of bovine embryo.


Assuntos
Bovinos , Senescência Celular/efeitos dos fármacos , Citoproteção/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Animais , Carnitina/farmacologia , Bovinos/embriologia , Bovinos/fisiologia , Células Cultivadas , Senescência Celular/genética , Embrião de Mamíferos , Desenvolvimento Embrionário/genética , Feminino , Glutationa/metabolismo , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Oócitos/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio/metabolismo
7.
J Reprod Dev ; 64(6): 489-494, 2018 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-30270255

RESUMO

Laminarin (LAM) is a ß-glucan oligomer known to possess biological activities such as anticancer and antioxidant effects. This study explored the influence of LAM supplementation on in vitro aged porcine oocytes and the underlying mechanisms behind this influence. We found that LAM delayed the aging process and improved the quality of aged oocytes. LAM supplementation enhanced the subsequent developmental competence of aged oocytes during the in vitro aging process. The blastocyst formation rate was significantly increased in aged oocytes treated with 20 µg/ml LAM compared to non-treated aged oocytes (45.3% vs. 28.7%, P < 0.01). The mRNA levels of apoptosis-related genes, B cell lymphoma-2-associated X protein (Bax) and Caspase-3, were significantly lower in blastocysts derived from the LAM-treated aged oocytes during the in vitro aging process. Furthermore, the level of intracellular reactive oxygen species was significantly decreased and that of glutathione was significantly increased in aged oocytes following LAM treatment. Mitochondrial membrane potential was increased, and the activities of caspase-3 and cathepsin B were significantly reduced in the LAM-treated aged oocytes compared with the non-treated aged oocytes. Taken together, these results suggest that LAM is beneficial for delaying the aging process in porcine oocytes.


Assuntos
Antioxidantes/farmacologia , Glucanos/farmacologia , Oócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Envelhecimento/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Glutationa/metabolismo , Técnicas de Maturação in Vitro de Oócitos , Oócitos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Suínos
8.
Zygote ; 24(2): 236-44, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25925489

RESUMO

We investigated the effect of human induced pluripotent stem cell (hiPS) medium on porcine somatic cell nuclear transfer and bovine in vitro fertilized early blastocysts, in comparison with North Carolina State University (NCSU)-37 medium and in vitro culture (IVC)-II medium. After 2 days of culture, the diameter of the portion of the blastocyst that was extruded from the zona pellucid dramatically differed between porcine blastocysts cultured in hiPS medium and those cultured in NCSU-37 medium (221.47 ± 38.94 µm versus 481.87 ± 40.61 µm, P < 0.01). Moreover, the diameter of the portion of the blastocyst significantly differed between bovine blastocysts cultured in hiPS medium and those cultured in IVC-II medium (150.30 ± 29.49 µm versus 195.58 ± 41.59 µm, P < 0.01). Furthermore, the total number of cells per porcine and bovine blastocyst was more than two-fold higher in blastocysts cultured in hiPS medium than in those cultured in NCSU-37 medium (44.33 ± 5.28 and 143.33 ± 16.05, P < 0.01) or IVC-II medium (172.12 ± 45.08 and 604.83 ± 242.64, P < 0.01), respectively. These results indicate that hiPS medium markedly improves the quality of porcine and bovine blastocysts.


Assuntos
Blastocisto/citologia , Meios de Cultura/farmacologia , Desenvolvimento Embrionário/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/citologia , Animais , Bovinos , Proliferação de Células/efeitos dos fármacos , Tamanho Celular/efeitos dos fármacos , Células Cultivadas , Meios de Cultura/química , Técnicas de Cultura Embrionária/métodos , Técnicas de Cultura Embrionária/veterinária , Feminino , Fertilização in vitro/veterinária , Humanos , Masculino , Microscopia de Fluorescência , Oócitos/citologia , Especificidade da Espécie , Suínos , Fatores de Tempo
9.
Biochem Biophys Res Commun ; 456(1): 156-61, 2015 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-25446119

RESUMO

In this study, we investigated the effects of the histone deacetylase inhibitor PXD101 (belinostat) on the preimplantation development of porcine somatic cell nuclear transfer (SCNT) embryos and their expression of the epigenetic markers histone H3 acetylated at lysine 9 (AcH3K9). We compared the in vitro developmental competence of SCNT embryos treated with various concentrations of PXD101 for 24h. Treatment with 0.5 µM PXD101 significantly increased the proportion of SCNT embryos that reached the blastocyst stage, in comparison to the control group (23.3% vs. 11.5%, P<0.05). We tested the in vitro developmental competence of SCNT embryos treated with 0.5 µM PXD101 for various amounts of times following activation. Treatment for 24h significantly improved the development of porcine SCNT embryos, with a significantly higher proportion of embryos reaching the blastocyst stage in comparison to the control group (25.7% vs. 10.6%, P<0.05). PXD101-treated SCNT embryos were transferred into two surrogate sows, one of whom became pregnant and four fetuses developed. PXD101 treatment significantly increased the fluorescence intensity of immunostaining for AcH3K9 in embryos at the pseudo-pronuclear and 2-cell stages. At these stages, the fluorescence intensities of immunostaining for AcH3K9 were significantly higher in PXD101-treated embryos than in control untreated embryos. In conclusion, this study demonstrates that PXD101 can significantly improve the in vitro and in vivo developmental competence of porcine SCNT embryos and can enhance their nuclear reprogramming.


Assuntos
Reprogramação Celular/efeitos dos fármacos , Reprogramação Celular/fisiologia , Embrião de Mamíferos/efeitos dos fármacos , Ácidos Hidroxâmicos/farmacologia , Técnicas de Transferência Nuclear , Sulfonamidas/farmacologia , Animais , Blastocisto/citologia , Epigênese Genética , Feminino , Fibroblastos/metabolismo , Inibidores de Histona Desacetilases/farmacologia , Histonas/metabolismo , Oócitos/citologia , Ovário/metabolismo , Gravidez , Suínos
10.
Anim Biotechnol ; 26(4): 292-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26158462

RESUMO

Wuzhishan miniature pig is one of the four most important pig breeds in China and has many major economic characteristics. Herein, we successfully used SCNT to clone Wuzhishan miniature pig. First, ear fibroblasts were isolated from a 2-year-old female Wuzhishan miniature piglet to be used as the donor cell. Second, good-quality COCs were selected from ovaries obtained from pigs at a local slaughterhouse and cultured. Mature eggs with the first polar body and ear fibroblasts were applied SCNT. Lastly, we in total produced 12 piglets with 7 piglets surviving to adults. Next, we used these pigs to test alloxan toxicity and to build T I D diabetes type. We know that diabetes mellitus is a chronic heterogeneous metabolic disease characterized by a high blood glucose level and abnormal insulin secretion. In this study, T I D (type I diabetes) was experimentally induced in cloned Wuzhishan miniature pigs with alloxan. In brief, an intravenous injection of alloxan (group B: 170 mg/kg, n = 3) was administered to pigs weighing between 27 and 39 kg. Sterile saline was administered to control pigs (n = 3). We determined the glycometabolism related index, performed an intravenous glucose tolerance test, and carried out immunohistochemistry experiments. There were no significant differences in body weight, blood glucose, and serum insulin in all groups, before treatment. The level of blood glucose was significantly higher (P < 0.05) in group B (12.18 ± 0.70 mmol/L) than in the control (2.93 ± 0.39 mmol/L). By contrast, the level of serum insulin was lower in group B (5.641 ± 0.573 µIU/mL) than in the control (7.578 ± 0.539 µIU/mL). Histological studies by hematoxylin and eosin (H&E) revealed a loss of ß-cells in the pancreas from pigs treated with 170 mg/kg alloxan. Immunolocalization studies showed a decrease in insulin reactivity in this treatment group as well. To conclude, our model holds promise in future studies of diabetes drug testing and islet xenotransplantation.


Assuntos
Clonagem Molecular/métodos , Diabetes Mellitus Experimental , Modelos Animais de Doenças , Técnicas de Transferência Nuclear , Porco Miniatura/genética , Aloxano , Animais , Glicemia/análise , Feminino , Fibroblastos/citologia , Insulina/sangue , Ilhotas Pancreáticas/química , Oócitos/citologia , Suínos , Testes de Toxicidade
11.
Biochem Biophys Res Commun ; 444(4): 638-43, 2014 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-24491539

RESUMO

Interspecies somatic cell nuclear transfer (iSCNT) is a promising method to clone endangered animals from which oocytes are difficult to obtain. Monomeric red fluorescent protein 1 (mRFP1) is an excellent selection marker for transgenically modified cloned embryos during somatic cell nuclear transfer (SCNT). In this study, mRFP-expressing rhesus monkey cells or porcine cells were transferred into enucleated porcine oocytes to generate iSCNT and SCNT embryos, respectively. The development of these embryos was studied in vitro. The percentage of embryos that underwent cleavage did not significantly differ between iSCNT and SCNT embryos (P>0.05; 71.53% vs. 80.30%). However, significantly fewer iSCNT embryos than SCNT embryos reached the blastocyst stage (2.04% vs. 10.19%, P<0.05). Valproic acid was used in an attempt to increase the percentage of iSCNT embryos that developed to the blastocyst stage. However, the percentages of embryos that underwent cleavage and reached the blastocyst stage were similar between untreated iSCNT embryos and iSCNT embryos treated with 2mM valproic acid for 24h (72.12% vs. 70.83% and 2.67% vs. 2.35%, respectively). These data suggest that porcine-rhesus monkey interspecies embryos can be generated that efficiently express mRFP1. However, a significantly lower proportion of iSCNT embryos than SCNT embryos reach the blastocyst stage. Valproic acid does not increase the percentage of porcine-rhesus monkey iSCNT embryos that reach the blastocyst stage. The mechanisms underling nuclear reprogramming and epigenetic modifications in iSCNT need to be investigated further.


Assuntos
Clonagem de Organismos/métodos , Proteínas Luminescentes/genética , Macaca mulatta/embriologia , Macaca mulatta/genética , Acetilação/efeitos dos fármacos , Animais , Eletroporação/métodos , Expressão Gênica , Inibidores de Histona Desacetilases/farmacologia , Histona Desacetilases/metabolismo , Histonas/metabolismo , Plasmídeos/administração & dosagem , Plasmídeos/genética , Ácido Valproico/farmacologia , Proteína Vermelha Fluorescente
12.
Food Chem Toxicol ; 185: 114445, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38311047

RESUMO

IsoliQuirtigenin (ILG) has been widely studied in somatic cells and tissues, but less in reproductive development. It is a kind of widely used food additive. In this study, it was found that ILG could significantly increase the levels of ROS,GSH and MMP in mouse oocytes (P < 0.01). In order to explore the cause of this phenomenon, it was found that the abnormal distribution of mitochondria and ATP synthesis levels were significantly increased (P < 0.05). At this time, we made a reasonable hypothesis that ILG affected mitochondrial function. In subsequent studies, it was found that the endogenous ROS accumulation level in mitochondria was significantly increased. After continuous RT-PCR screening, it was found that the expression of Nrf2 was significantly inhibited (P < 0.01). Its upstream and downstream FOXO3 GPX1, CAT, SOD2, SIRT1 gene also appear different degree of significant change (P < 0.05), in which the lower expression of NADP + (P < 0.05) illustrates the mitochondrial ATP synthesis electronic chain were suppressed, it also has the reason, By inhibiting electron chain and ATP synthesis, ILG leads to oocyte apoptosis and initiation of autophagy, reducing oocyte and its subsequent developmental potential.


Assuntos
Chalcona/análogos & derivados , Glucosídeos , Doenças Mitocondriais , Fator 2 Relacionado a NF-E2 , Camundongos , Animais , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Técnicas de Maturação in Vitro de Oócitos , Espécies Reativas de Oxigênio/metabolismo , Oócitos , Trifosfato de Adenosina/metabolismo
13.
Theriogenology ; 218: 8-15, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38290232

RESUMO

To investigate the effects of limonin (Lim) on oxidative stress and early apoptosis in bovine oocytes during in vitro maturation (IVM), different concentrations of Lim (0, 10, 20, 50 µmol/L) were added to bovine IVM medium. Oocyte maturation rates and development 24 h after in vitro fertilization (IVF) were examined to determine the optimal Lim concentration. The optimal Lim concentration was added to the IVM medium, and 0 µmol/L Lim was used as the control. Immunofluorescence staining was used to detect the abnormal rate of spindle assembly, reactive oxygen species (ROS), glutathione (GSH), mitochondrial membrane potential (MMP) levels, mitochondrial distribution, and the fluorescence intensity of cathepsin B (CB)-active LC3 protein. RT‒qPCR was used to detect the mRNA expression levels of antioxidant-, apoptosis- and autophagy-related genes in oocytes. The total number of blastocysts and the proportion of apoptotic cells among blastocysts were detected. The results showed that the PBI ejection rate, cleavage rate and blastocyst rate of bovine oocytes in the 20 µmol/L Lim group were significantly higher than those in the control group (P < 0.05). Compared with those in the control group, ROS levels, abnormal mitochondrial distribution, the proportion of abnormal spindle assembly, CB activity and LC3 protein fluorescence intensity of oocytes in the 20 µmol/L Lim group were significantly decreased (P < 0.05), and GSH and MMP levels were significantly increased (P < 0.05). The expression of antioxidant genes (Prdx3, Prdx6, Sirt1) and antiapoptotic genes (Bcl-xl, Survivin) were significantly upregulated (P < 0.05), and the expression levels of proapoptotic genes (Caspase-4, BAX) and autophagy-related genes (LC3) were significantly downregulated (P < 0.05). The total number of cells among in vitro fertilized embryos was significantly increased (P < 0.05), and the apoptosis rate of blastocysts was significantly decreased (P < 0.05). Here, we show that Lim exerts positive effects on bovine oocyte IVM by regulating REDOX homeostasis, reducing spindle damage and enhancing mitochondrial function during IVM, thereby inhibiting oocyte apoptosis and autophagy.


Assuntos
Antioxidantes , Limoninas , Animais , Bovinos , Antioxidantes/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Limoninas/metabolismo , Limoninas/farmacologia , Oócitos/fisiologia , Estresse Oxidativo , Glutationa/metabolismo , Blastocisto/fisiologia , Apoptose , Desenvolvimento Embrionário
14.
Theriogenology ; 218: 89-98, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38308957

RESUMO

After ovulation, senescent oocytes inevitably experience reduced quality and defects in embryonic development. Apigenin (API) is a flavonoid with a wide range of pharmacological effects. Therefore, this study examined the protective effects of API on the quality of porcine oocytes during in-vitro ageing and the underlying mechanisms. The results showed that API treatment could reduce the activation rate after aging for 48 h. In addition, API significantly reduced reactive oxygen species, abnormal distribution of mitochondria, early apoptosis in ageing oocytes, increased glutathione, and mitochondrial adenosine triphosphate levels in ageing oocytes. Importantly, API increased the embryonic development rate in aged oocytes. We also examined molecular changes, finding decreased sirtuin 1 expression in in-vitro postovulatory oocytes, but API reversed this effect. Our results suggest that API attenuates the deterioration of oocyte quality during in-vitro ageing, possibly by reducing oxidative stress through the upregulation of sirtuin 1.


Assuntos
Apigenina , Sirtuína 1 , Feminino , Animais , Suínos , Sirtuína 1/genética , Sirtuína 1/metabolismo , Apigenina/farmacologia , Apigenina/metabolismo , Regulação para Cima , Senescência Celular/fisiologia , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Oócitos/fisiologia
15.
Theriogenology ; 203: 53-60, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36972665

RESUMO

Strategies for improving the quality of oocytes have important theoretical and practical significance for increasing the efficiency of livestock breeding. In this respect, the accumulation of reactive oxygen species (ROS) is a major factor affecting the development of oocytes and embryos. This study investigated the effects of Dendrobium nobile extract (DNE) on the in vitro maturation of bovine oocytes and embryonic development after IVF. DNE is an extract from Dendrobium rhizomes that contains alkaloids with anti-inflammatory, anti-cancer and anti-ageing functions. Various concentrations of DNE (0, 5, 10, 20 and 50 µmol/L) were added during oocyte maturation in vitro, and we found that 10 µmol/L of DNE remarkably increased the oocyte maturation rate, the subsequent blastocyst formation rate and embryo quality. Further, we found that DNE treatment decreased the frequency of spindle/chromosome defects and ROS and increased the oocyte glutathione and mitochondrial membrane potential in oocytes. Moreover, DNE upregulated the expression of oxidative stress-related genes (Sirt1, Sirt2, Sirt3 and Sod1) in oocytes and apoptosis-related genes (Caspase-3, Caspase-4, Bax, Bcl-xl and Survivin) in blastocysts. These results suggest that DNE supplementation can promote oocyte maturation and subsequent embryonic development by regulating redox reactions and inhibiting embryonic apoptosis.


Assuntos
Técnicas de Maturação in Vitro de Oócitos , Oócitos , Animais , Bovinos , Espécies Reativas de Oxigênio/metabolismo , Técnicas de Maturação in Vitro de Oócitos/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Oócitos/fisiologia , Desenvolvimento Embrionário , Blastocisto/fisiologia
16.
Zool Res ; 43(2): 275-284, 2022 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-35238185

RESUMO

Environmental temperature serves as a major driver of adaptive changes in wild organisms. To discover the mechanisms underpinning cold tolerance in domestic animals, we sequenced the genomes of 28 cattle from warm and cold areas across China. By characterizing the population structure and demographic history, we identified two genetic clusters, i.e., northern and southern groups, as well as a common historic population peak at 30 kilo years ago. Genomic scan of cold-tolerant breeds determined potential candidate genes in the thermogenesis-related pathways that were under selection. Specifically, functional analysis identified a substitution of PRDM16 (p.P779L) in northern cattle, which maintains brown adipocyte formation by boosting thermogenesis-related gene expression, indicating a vital role of this gene in cold tolerance. These findings provide a basis for genetic variation in domestic cattle shaped by environmental temperature and highlight the role of reverse mutation in livestock species.


Assuntos
Metagenômica , Termogênese , Animais , Bovinos/genética , China , Temperatura Baixa , Genoma , Termogênese/genética
17.
Yi Chuan ; 33(11): 1219-24, 2011 Nov.
Artigo em Zh | MEDLINE | ID: mdl-22120077

RESUMO

Annealing control primer (ACP) system was applied to find candidate genes related to lipidosis in muscle of Yanbian yellow cattle by screening differentially expressed genes (DEGs) in Longissimus dorsi, which had significant difference on intramuscular fat (IMF) content. Thirty steers, aged at 28 month-bullocks were selected to measure the IMF content in L. dorsi. Two groups of bullocks (three heads per group) with the highest and the lowest contents of IMF were selected to build a RNA pool, and DEGs of two groups were analyzed by ACP system. Twelve DEGs were identified and sequenced by amplification with 20 arbitrary primers (fragment sizes were 200-890 bp). In these genes, eight were already known as functional groups of cytoskeleton, cytokine signal transduction, protein synthesis, energy metabolism, and others, four were unknown. All the 12 ESTs were screened by ACP system, which may participated in regulating on lipidosis in muscle. This study established a foundation for further screening of lipidosis related genes.


Assuntos
Bovinos/genética , Clonagem Molecular , Expressão Gênica , Animais , Bovinos/metabolismo , Gorduras/metabolismo , Masculino , Músculo Esquelético/metabolismo , Proteínas/genética , Proteínas/metabolismo
18.
Theriogenology ; 143: 64-73, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31837632

RESUMO

Aging oocytes undergo various molecular, cellular, and biochemical changes. Aging of oocytes results in reduced embryo developmental capacity and blastocyst quality, which is thought to be caused partly by the accumulation of reactive oxygen species (ROS). This study aimed to determine the effect of l-carnitine (LC) on the development of embryos formed from aged oocytes in vitro. The development and quality of the blastocysts in the LC-treated group were significantly higher than those in the untreated aged group after in vitro fertilization (IVF). In addition, after LC treatment, the level of intracellular ROS in aged group significantly decreased, and glutathione (GSH) levels significantly increased compared with those in the untreated aged group. There was no significant difference in the mitochondrial membrane potential among the three groups. Moreover, ROS could induce autophagy and LC3 antibody was widely used as a marker for detecting autophagy. The fluorescence intensity of LC3 in the aged group was significantly higher than that of LC3 in the LC-treated group. Furthermore, Real-time reverse transcriptase-polymerase chain reaction showed that the mRNA levels of antioxidation genes GPX4 and SOD1 were significantly higher in embryos from LC-treated group than in those from the untreated aged group. In summary, our results indicated that LC can improve the developmental capacity of embryos from aging oocytes in vitro by reducing oxidative stress.


Assuntos
Carnitina/farmacologia , Bovinos/embriologia , Técnicas de Cultura Embrionária/veterinária , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/fisiologia , Animais , Meios de Cultura , Feminino , Fertilização in vitro/veterinária , Estresse Oxidativo
19.
Theriogenology ; 151: 66-73, 2020 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-32311602

RESUMO

Zearalenone (ZEA), which is produced by several fusarium mycotoxins, is found in animal feed and food products, and can exert estrogen-like activity. Melatonin (MT) is emerging as a supplement that can fight the toxic effects of mycotoxins. With a variety of physiological functions that play crucial roles in the development of animal germ cells and embryos, melatonin regulates circadian rhythms and has an anti-inflammatory and anti-oxidative role. This study investigated the protective effects of melatonin against ZEA in porcine early embryonic development. Our results showed that ZEA adversely affected this development, while melatonin supplementation ameliorated the toxic effects. ZEA exposure increased oxidative stress and impaired mitochondrial function, which may affect blastocyst formation. Moreover, we found that ZEA exposure promotes apoptosis, DNA damage, and autophagy in porcine blastocysts. The toxic effects of ZEA on early embryos may be the result of oxidative stress-mediated early apoptosis, while melatonin treatment significantly improved these phenotypes in ZEA-exposed porcine early embryos. Taken together, our results indicate that melatonin has a protective effect on defects caused by ZEA during early porcine embryonic development.


Assuntos
Blastocisto/efeitos dos fármacos , Melatonina/farmacologia , Suínos/embriologia , Zearalenona/toxicidade , Animais , Apoptose/efeitos dos fármacos , Técnicas de Cultura Embrionária , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/fisiologia , Partenogênese , Espécies Reativas de Oxigênio
20.
J Thorac Dis ; 12(12): 7272-7280, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33447416

RESUMO

BACKGROUND: The incision protective sleeve can protect incisions and help to establish an operating port and thus has been widely applied in thoracic surgeries. However, its other utilities are often neglected. This article explores the additional functions and placement techniques of incision protective sleeves in video-assisted transthoracic surgery (VATS). METHODS: Operators with different surgical experience were divided into three groups: resident group, attending surgeon group, and professor group. Each group independently chose one of the four surgical maneuvers, and the incision protective sleeve was placed during the operation. Up to 200 operations were randomly selected in each group, and the patients' gender, age, incision site, incision length, the operator's experience, and the time and technique of incision protective sleeve placement were recorded. CT was performed to measure the thickness of chest wall and the width of intercostal spaces. Data were analyzed using SPSS 21.0 software package. Multivariate linear regression analysis was performed was performed for the time required for incision protective sleeve placement. RESULTS: The operator's experience was inversely related to the time required for incision protective sleeve placement, width of intercostal spaces was negatively correlated with operative time, chest wall thickness and incision length were positively correlated with operative time. Among the maneuvers, incision protective sleeve placement skills were significant different. CONCLUSIONS: The placement of the incision protective sleeve for VATS is affected by multiple factors, which are not only related to the patient's condition, chest wall thickness and intercostal space, but also closely related to the operator's experience and the manipulation adopted.

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