RESUMO
This study investigated the effect of the flavonoid-based compound isorhamnetin (ISO) on maturation and developmental competence in oxidative stress-exposed porcine oocytes in vitro. Treatment with 2 µM ISO (2 ISO) increases the developmental rate of oxidative stress-exposed porcine oocytes during in vitro maturation (IVM). The glutathione level and mRNA expression of antioxidant-related genes (NFE2L2 and SOD2) were increased in the 2 ISO-treated group, whereas the reactive oxygen species level was decreased. Treatment with 2 ISO increased mRNA expression of a cumulus cell expansion-related gene (SHAS2) and improved chromosomal alignment. mRNA expression of maternal genes (CCNB1, MOS, BMP15 and GDF9) and mitogen activated protein kinase (MAPK) activity were increased in the 2 ISO-treated group. The total cell number per blastocyst and percentage of apoptotic cells were increased and decreased in the 2 ISO-treated group, respectively. Treatment with 2 ISO increased mRNA expression of development-related genes (SOX2, NANOG, and POU5F1) and anti-apoptotic genes (BCL2L1 and BIRC5) and decreased that of pro-apoptotic genes (CASP3 and FAS). These results demonstrate that 2 ISO improves the quality of porcine oocytes by protecting them against oxidative stress during IVM and enhances subsequent embryo development in vitro. Therefore, we propose that ISO is a useful supplement for IVM of porcine oocytes.
Assuntos
Desenvolvimento Embrionário , Técnicas de Maturação in Vitro de Oócitos , Oócitos , Estresse Oxidativo , Animais , Blastocisto/metabolismo , Desenvolvimento Embrionário/efeitos dos fármacos , Técnicas de Maturação in Vitro de Oócitos/métodos , Oócitos/efeitos dos fármacos , Oócitos/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , SuínosRESUMO
Climate change-related drought stress is expected to shift carbon partitioning toward volatile organic compound (VOC) biosynthesis. The effect of drought stress on VOC synthesis remains unknown in several tree species. Therefore, we exposed eastern white pine (Pinus strobus) plants to severe drought for 32 days and performed physiological analysis (chlorophyll content, leaf water content, and root/shoot index), biochemical analysis (non-structural carbohydrates, proline, lipid peroxidation, and antioxidant assay), and total root VOC analysis. Drought stress decreased the relative water and soil moisture contents. Root proline accumulation and antioxidant activity increased significantly, whereas leaf chlorophyll synthesis and fresh weight decreased significantly in drought-treated plants. A non-significant increase in sugar accumulation (leaves and roots), proline accumulation (leaves), antioxidant activity (leaves), and lipid peroxidation (leaves and roots) was observed in drought-treated plants. Drought stress caused a non-significant decline in root/shoot ratio and starch accumulation (leaves and roots) and caused a significant increase in root abscisic acid content. Drought-treated plants showed an increase in overall monoterpene synthesis (16%) and decline in total sesquiterpene synthesis (3%). Our findings provide an overall assessment of the different responses of VOC synthesis to severe water deficit that may help unravel the molecular mechanisms underlying drought tolerance in P. strobus.
RESUMO
Our previous studies have already revealed that ß-cryptoxanthin (BCX), hesperetin (HES), and icariin (ICA) antioxidants are effective for in vitro maturation (IVM) of porcine oocytes. In this study, we investigated which of BCX, HES, or ICA was more effective for IVM of porcine oocytes. The antioxidant properties were assessed with aged porcine oocytes and embryos by comparing 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH), reducing power, and H2O2 scavenging activity assays. The chemical assay results demonstrated that BCX had a greater DPPH scavenging activity and reducing power than HES and ICA, compared with controls. However, the H2O2 scavenging activity of the antioxidants was similar when tested at the optimal concentrations of 1 µM BCX (BCX-1), 100 µM HES (HES-100), and 5 µM ICA (ICA-5). The biological assay results showed that BCX-1 treatment was more effective in inducing a significant reduction in reactive oxygen species (ROS), improving glutathione levels, and increasing the expression of antioxidant genes. In addition, BCX-1 inhibited apoptosis by increasing the expression of anti-apoptotic genes and decreasing pro-apoptotic genes in porcine parthenogenetic blastocysts. BCX-1 also significantly increased the blastocyst formation rate compared with the ageing control group, HES-100 and ICA-5. This study demonstrates that damage from ROS produced during oocyte ageing can be prevented by supplementing antioxidants into the IVM medium, and BCX may be a potential candidate to improve assisted reproductive technologies.
Assuntos
Antioxidantes , Técnicas de Maturação in Vitro de Oócitos , Animais , Antioxidantes/metabolismo , Bioensaio , Blastocisto/metabolismo , Desenvolvimento Embrionário , Peróxido de Hidrogênio/farmacologia , Técnicas de Maturação in Vitro de Oócitos/métodos , Oócitos , Espécies Reativas de Oxigênio/metabolismo , SuínosRESUMO
This study investigated the effect of the antioxidant dieckol, a component of Ecklonia cava, on maturation and developmental competence of porcine oocytes exposed to oxidative stress in vitro. Oocytes were matured in in vitro maturation (IVM) medium containing various concentrations of dieckol. The blastocyst formation rate was highest in the 0.5 µM dieckol-treated (0.5 DEK) group. The reactive oxygen species level was decreased, and the level of glutathione and expression of antioxidant genes (NFE2L, SOD1, and SOD2) at metaphase II were increased in the 0.5 DEK group. Abnormal spindle organization and chromosome misalignment were prevented in the 0.5 DEK group. Expression of maternal markers (CCNB1 and MOS) and activity of p44/42 mitogen-activated protein kinase were increased in the 0.5 DEK group. After parthenogenetic activation, the total number of cells per blastocyst was increased and the percentage of apoptotic cells was decreased in the 0.5 DEK group. Expression of development-related genes (CX45, CDX2, POU5F1, and NANOG), antiapoptotic genes (BCL2L1 and BIRC5), and a proapoptotic gene (CASP3) were altered in the 0.5 DEK group. These results indicate that the antioxidant dieckol improves IVM and subsequent development of porcine oocytes and can be used to improve the quality of oocytes under peroxidation experimental conditions.
Assuntos
Antioxidantes/farmacologia , Benzofuranos/farmacologia , Desenvolvimento Embrionário/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Partenogênese/efeitos dos fármacos , Animais , Antioxidantes/administração & dosagem , Benzofuranos/administração & dosagem , Blastocisto/citologia , Posicionamento Cromossômico/efeitos dos fármacos , Relação Dose-Resposta a Droga , Técnicas de Cultura Embrionária , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Glutationa/metabolismo , Técnicas de Maturação in Vitro de Oócitos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Meiose , Oócitos/metabolismo , Phaeophyceae/química , Espécies Reativas de Oxigênio/metabolismo , Fuso Acromático/efeitos dos fármacos , Fuso Acromático/ultraestrutura , SuínosRESUMO
OBJECTIVE: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. METHODS: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. RESULTS: Treatment with 5 µM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. CONCLUSION: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.
RESUMO
Chronic inflammation has been linked to colitis-associated colorectal cancer in humans. The human symbiont enterotoxigenic Bacteroides fragilis (ETBF), a pro-carcinogenic bacterium, has the potential to initiate and/or promote colorectal cancer. Antibiotic treatment of ETBF has shown promise in decreasing colonic polyp formation in murine models of colon cancer. However, there are no reported natural products that have shown efficacy in decreasing polyp burden. In this study, we investigated the chemopreventive effects of oral administration of zerumbone in ETBF-colonized mice with azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced tumorigenesis. Zerumbone significantly reduced the severity of disease activity index (DAI) scores as well as several parameters of colonic inflammation (i.e., colon weight, colon length, cecum weight and spleen weight). In addition, inflammation of the colon and cecum as well as hyperplasia was reduced. Zerumbone treatment significantly inhibited colonic polyp numbers and prevented macroadenoma progression. Taken together, these findings suggest that oral treatment with zerumbone inhibited ETBF-promoted colon carcinogenesis in mice indicating that zerumbone could be employed as a promising protective agent against ETBF-mediated colorectal cancer.
Assuntos
Bacteroides fragilis/patogenicidade , Neoplasias do Colo/prevenção & controle , Substâncias Protetoras/uso terapêutico , Sesquiterpenos/uso terapêutico , Administração Oral , Animais , Azoximetano/toxicidade , Peso Corporal/efeitos dos fármacos , Carcinogênese/efeitos dos fármacos , Colite/complicações , Colite/microbiologia , Colite/patologia , Neoplasias do Colo/induzido quimicamente , Neoplasias do Colo/patologia , Sulfato de Dextrana/toxicidade , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Substâncias Protetoras/farmacologia , Sesquiterpenos/farmacologia , Índice de Gravidade de DoençaRESUMO
The azoxymethane (AOM)/dextran sulfate sodium (DSS) murine model is commonly used to study colitis-associated cancer. The human commensal bacterium, enterotoxigenic Bacteroides fragilis (ETBF) secretes the Bacteroides fragilis toxin (BFT) which is necessary and sufficient to cause colitis. We report that BALB/c mice infected with WT-ETBF and administered three cycles of AOM/DSS developed numerous, large-sized polyps predominantly in the colorectal region. In addition, AOM/DSS-treated BALB/c mice orally inoculated with wild-type nontoxigenic Bacteroides fragilis (WT-NTBF) overexpressing bft (rETBF) developed numerous polyps whereas mice infected with WT-NTBF overexpressing a biologically inactive bft (rNTBF) did not promote polyp formation. Unexpectedly, the combination of AOM+ETBF did not induce polyp formation whereas ETBF+DSS did induce polyp development in a subset of BALB/c mice. In conclusion, WT-ETBF promoted polyp development in AOM/DSS murine model with increased colitis in BALB/c mice. The model described herein provides an experimental platform for understanding ETBF-induced colonic tumorigenesis and studying colorectal cancer in wild-type mice.
Assuntos
Infecções por Bacteroides/patologia , Carcinogênese/genética , Colite/patologia , Neoplasias Colorretais/patologia , Animais , Azoximetano/toxicidade , Toxinas Bacterianas/toxicidade , Infecções por Bacteroides/induzido quimicamente , Infecções por Bacteroides/complicações , Infecções por Bacteroides/microbiologia , Bacteroides fragilis/patogenicidade , Carcinogênese/induzido quimicamente , Colite/induzido quimicamente , Colite/complicações , Colite/microbiologia , Colo/efeitos dos fármacos , Colo/patologia , Neoplasias Colorretais/induzido quimicamente , Neoplasias Colorretais/complicações , Neoplasias Colorretais/microbiologia , Sulfato de Dextrana/toxicidade , Modelos Animais de Doenças , Humanos , Metaloendopeptidases/toxicidade , Camundongos , Pólipos/induzido quimicamenteRESUMO
Enterotoxigenic Bacteroides fragilis (ETBF) is human intestinal commensal bacterium and a potent initiator of colitis through secretion of the metalloprotease Bacteroides fragilis toxin (BFT). BFT induces cleavage of E-cadherin in colon cells, which subsequently leads to NF-κB activation. Zerumbone is a key component of the Zingiber zerumbet (L.) Smith plant and can exhibit anti-bacterial and anti-inflammatory effects. However, whether zerumbone has anti-inflammatory effects in ETBF-induced colitis remains unknown. The aim of this study was to determine the anti-inflammatory effect of orally administered zerumbone in a murine model of ETBF infection. Wild-type C57BL/6 mice were infected with ETBF and orally administered zerumbone (30 or 60 mg/kg) once a day for 7 days. Treatment of ETBF-infected mice with zerumbone prevented weight loss and splenomegaly and reduced colonic inflammation with decreased macrophage infiltration. Zerumbone treatment significantly decreased expression of IL-17A, TNF-α, KC, and inducible nitric oxide synthase (iNOS) in colonic tissues of ETBF-infected mice. In addition, serum levels of KC and nitrite was also diminished. Zerumbone-treated ETBF-infected mice also showed decreased NF-κB signaling in the colon. HT29/C1 colonic epithelial cells treated with zerumbone suppressed BFT-induced NF-κB signaling and IL-8 secretion. However, BFT-mediated E-cadherin cleavage was unaffected. Furthermore, zerumbone did not affect ETBF colonization in mice. In conclusion, zerumbone decreased ETBF-induced colitis through inhibition of NF-κB signaling.
Assuntos
Antibacterianos/uso terapêutico , Infecções por Bacteroides/tratamento farmacológico , Bacteroides fragilis , Colite/tratamento farmacológico , NF-kappa B/antagonistas & inibidores , Sesquiterpenos/uso terapêutico , Animais , Toxinas Bacterianas , Infecções por Bacteroides/imunologia , Bacteroides fragilis/metabolismo , Caderinas/metabolismo , Colite/imunologia , Colo/efeitos dos fármacos , Colo/imunologia , Colo/fisiopatologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/imunologia , Células Epiteliais/patologia , Células HT29 , Humanos , Interleucina-17/metabolismo , Interleucina-8/sangue , Metaloendopeptidases , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico Sintase Tipo II/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Optimization of culture conditions is important to improve oocyte maturation and subsequent embryo development. In particular, this study analyzed the effects of increasing concentrations of PIO in the maturation medium on spindle formation and chromosome alignment, glutathione, and intracellular ROS levels and expression of selected genes related to maternal markers, apoptosis, and lipid metabolism. The percentage of oocytes displaying normal spindle formation and chromosome alignment was higher in the 1 µM PIO (1 PIO)-treated group than in the control group. The glutathione level was significantly higher in the 1 PIO-treated group than in the control group, while the reactive oxygen species level did not differ. Expression of maternal marker (MOS and GDF9), antiapoptotic (BIRC5), and lipid metabolism-related (ACADS, CPT2, SREBF1, and PPARG) genes was higher in the 1 PIO-treated group than in the control group, while expression of a proapoptotic gene (CASP3) was lower. The blastocyst formation rate and the percentage of blastocysts that reached at least the hatching stage on Days 6 and 7, and the percentage of blastocysts containing more than 128 cells were significantly higher in the 1 PIO-treated group than in the control group. These results indicate that PIO treatment during in vitro maturation improves porcine oocyte maturation and subsequent parthenogenetic embryo development mainly by enhancing lipid metabolism and antioxidant defense in oocytes.
Assuntos
Embrião de Mamíferos/embriologia , Desenvolvimento Embrionário/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Oócitos/metabolismo , Partenogênese/efeitos dos fármacos , Pioglitazona/farmacologia , Animais , Embrião de Mamíferos/citologia , SuínosRESUMO
BACKGROUND: Tracheal intubation is closely associated with increases in intraocular pressure (IOP); however, the effects of double-lumen tube (DLT) intubation on IOP have not been validated. Systemic hypertension (HTN) is another factor that may increase IOP. In this study, we observed differences in IOP increases between DLT and singlelumen tube (SLT) intubation, and evaluated the influence of underlying HTN during rapid sequence induction. METHODS: Sixty-eight patients were allocated into one of the following group: SLT/without HTN (n = 17), SLT/HTN (n = 17), DLT/without HTN (n = 17), and DLT/HTN (n = 17). An SLT was inserted for orthopedic or gynecological surgery, and a DLT was inserted for lung surgery after rapid sequence induction using succinylcholine. IOP was measured before anesthetic induction and until 10 min after intubation using a handheld tonometer (Tono-Pen AVIA®). RESULTS: In the DLT/without HTN and DLT/HTN groups, the maximum increases in IOPs after tracheal intubation were 7.9 and 12.2 mmHg, respectively, compared to baseline. In the SLT/without HTN and SLT/HTN groups, the maximum increases were 5.0 and 4.9 mmHg, respectively, compared to baseline. In comparisons between patients with and without underlying HTN, the values of IOPs were comparable. CONCLUSIONS: Tracheal intubation with a DLT is associated with more increases in IOPs than with an SLT in rapid sequence induction. Well-controlled underlying hypertension did not increase IOP during tracheal intubation.