scholarly journals Η επίδραση καροτενοειδών του κρόκου του ήμερου (Crocus Sativus Linneaus) σε ποιοτικά χαρακτηριστικά του σπέρματος του ταύρου και στην in vitro γονιμοποίηση

2015 ◽  
Author(s):  
Βασιλική Σαπανίδου
Keyword(s):  
Reactive Oxygen Species ◽  
Reactive Oxygen ◽  
Crocus Sativus ◽  
Oxygen Species ◽  
Crocus Sativus L

Τα κύτταρα (σπερματοζωάρια, ωοκύτταρα, έμβρυα) που συμμετέχουν στην in vitro γονιμοποίηση υφίστανται έντονο οξειδωτικό στρες κατά τη διάρκεια των χειρισμών και της καλλιέργειάς τους, καθώς οι συνθήκες κάτω από τις οποίες πραγματοποιούνται οι χειρισμοί χαρακτηρίζονται από υψηλή συγκέντρωση οξυγόνου. Το οξειδωτικό στρες αποτελεί μία από τις κυριότερες αιτίες της υποβάθμισης της ποιότητας του σπέρματος και των χαμηλών ποσοστών επιτυχίας της in vitro γονιμοποίησης. Υψηλές συγκεντρώσεις δραστικών μορφών οξυγόνου (reactive oxygen species, ROS) θίγουν τη δομική και λειτουργική ακεραιότητα των γαμετών. Ωστόσο, μικρές και ελεγχόμενες συγκεντρώσεις ROS είναι απαραίτητες, καθώς ρυθμίζουν βασικές φυσιολογικές διεργασίες της αναπαραγωγής, όπως είναι η ενεργοποίηση των σπερματοζωαρίων και η συγχώνευση των γαμετών. Τα τελευταία χρόνια η επιστημονική κοινότητα έχει στραφεί στην προσθήκη αντιοξειδωτικών παραγόντων στα υποστρώματα της in vitro παραγωγής εμβρύων προκειμένου να αποσοβήσουν τον κίνδυνο που εγκυμονούν οι υψηλές συγκεντρώσεις ROS στη γονιμοποιητική ικανότητα του σπέρματος και στα ποσοστά επιτυχίας της in vitro γονιμοποίησης. Οι αντιοξειδωτικές ιδιότητες του εκχυλίσματος των στύλων του κρόκου του ήμερου (Crocus. sativus L.) έχουν μελετηθεί τόσο in vivo, όσο και in vitro. Έτσι, η πιθανή αντιοξειδωτική δράση του κρόκου του ήμερου έχει ιδιαίτερο ενδιαφέρον να μελετηθεί σε κύτταρα που υφίστανται έντονο οξειδωτικό στρες, όπως αυτά που συμμετέχουν στην in vitro γονιμοποίηση. Στη διατριβή αυτή διερευνάται η επίδραση δύο κύριων βιολογικώς ενεργών συστατικών του κρόκου του ήμερου, της κροκίνης και της κροκετίνης, στη γονιμοποιητική ικανότητα του κατεψυγμένου σπέρματος ταύρου in vitro. Για το σκοπό της αυτό χρησιμοποιήθηκαν κατεψυγμένα σπερματοζωάρια ταύρου και ωοκύτταρα από ωοθήκες αγελάδων, ύστερα από τη μεταθανάτια συλλογή τους. Ελέγχθηκε η επίδραση 3 διαφορετικών συγκεντρώσεων κροκίνης (0,5mM, 1mM και 2mM) και κροκετίνης (1μΜ, 2,5μΜ και 5μΜ) σε ποιοτικές παραμέτρους του σπέρματος, όπως η κινητικότητα, η ζωτικότητα, η ενεργοποίηση/αντίδραση ακροσώματος, η ακεραιότητα του DNA, τα ενδοκυτταρικά επίπεδα της ρίζα του ανιόντος υπεροξειδίου και του υπεροξειδίου του υδρογόνου, το ποσοστό μετάθεσης της φωσφατιδυλοσερίνης στην εξωτερική κυτταρική μεμβράνη και ο βαθμός της λιπιδικής υπεροξείδωσης των κυτταρικών μεμβρανών. Η αξιολόγηση των παραπάνω παραμέτρων πραγματοποιήθηκε σε 3 χρονικά σημεία: 0 ώρα, 2 ώρες και 4 ώρες επώασης. Η επιβεβαίωση της βελτίωσης της γονιμοποιητικής ικανότητας του σπέρματος από την παρουσία των δύο καροτενοειδών έγινε με την τροποποίηση του υποστρώματος της in vitro γονιμοποίησης και τον έλεγχο του ποσοστού αυλάκωσης και των παραγόμενων βλαστικών κύστεων, παρουσία αρνητικού μάρτυρα. Από την εκτίμηση των αποτελεσμάτων προκύπτει ότι η επώαση του σπέρματος παρουσία της κροκίνης (1mM) και της κροκετίνης (2,5μΜ) διασφαλίζει ελεγχόμενα ποσά ROS και λιπιδικών υδροϋπεροξειδίων. Ο αντιοξειδωτικός αυτός ρόλος προστατεύει, σύμφωνα με τα αποτελέσματα, την κινητικότητα, τη ζωτικότητα και την ακεραιότητα του DNA, ιδιότητες που συνδέονται άρρηκτα με τη γονιμοποιητική ικανότητα του σπέρματος. Η τελευταία επιβεβαιώνεται μέσω της σημαντικής αύξησης του ποσοστού των σπερματοζωαρίων με αντίδραση ακροσώματος και των παραγόμενων βλαστικών κύστεων, συγκριτικά με το μάρτυρα.

scholarly journals Targeting autophagy enhances atezolizumab-induced mitochondria-related apoptosis in osteosarcoma

2021 ◽  
Vol 12 (2) ◽  
Author(s):  
Zhuochao Liu ◽  
Hongyi Wang ◽  
Chuanzhen Hu ◽  
Chuanlong Wu ◽  
Jun Wang ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Antitumor Immunity ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Specific Monoclonal Antibody ◽  
Osteosarcoma Cells ◽  
Jnk Pathway ◽  
In Vivo Experiments

AbstractIn this study, we identified the multifaceted effects of atezolizumab, a specific monoclonal antibody against PD-L1, in tumor suppression except for restoring antitumor immunity, and investigated the promising ways to improve its efficacy. Atezolizumab could inhibit the proliferation and induce immune-independent apoptosis of osteosarcoma cells. With further exploration, we found that atezolizumab could impair mitochondria of osteosarcoma cells, resulting in increased release of reactive oxygen species and cytochrome-c, eventually leading to mitochondrial-related apoptosis via activating JNK pathway. Nevertheless, the excessive release of reactive oxygen species also activated the protective autophagy of osteosarcoma cells. Therefore, when we combined atezolizumab with autophagy inhibitors, the cytotoxic effect of atezolizumab on osteosarcoma cells was significantly enhanced in vitro. Further in vivo experiments also confirmed that atezolizumab combined with chloroquine achieved the most significant antitumor effect. Taken together, our study indicates that atezolizumab can induce mitochondrial-related apoptosis and protective autophagy independently of the immune system, and targeting autophagy is a promising combinatorial approach to amplify its cytotoxicity.

scholarly journals Plant Extracts and Reactive Oxygen Species as Two Counteracting Agents with Anti- and Pro-Obesity Properties

2019 ◽  
Vol 20 (18) ◽  
pp. 4556 ◽  
Author(s):  
Hanna Zielinska-Blizniewska ◽  
Przemyslaw Sitarek ◽  
Anna Merecz-Sadowska ◽  
Katarzyna Malinowska ◽  
Karolina Zajdel ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Plant Extracts ◽  
Complex Disease ◽  
Reactive Oxygen ◽  
Complementary Therapy ◽  
Biologically Active ◽  
Mitochondrial Activity ◽  
Oxygen Species

Obesity is a complex disease of great public health significance worldwide: It entails several complications including diabetes mellitus type 2, cardiovascular dysfunction and hypertension, and its prevalence is increasing around the world. The pathogenesis of obesity is closely related to reactive oxygen species. The role of reactive oxygen species as regulatory factors in mitochondrial activity in obese subjects, molecules taking part in inflammation processes linked to excessive size and number of adipocytes, and as agents governing the energy balance in hypothalamus neurons has been examined. Phytotherapy is the traditional form of treating health problems using plant-derived medications. Some plant extracts are known to act as anti-obesity agents and have been screened in in vitro models based on the inhibition of lipid accumulation in 3T3-L1 cells and activity of pancreatic lipase methods and in in vivo high-fat diet-induced obesity rat/mouse models and human models. Plant products may be a good natural alternative for weight management and a source of numerous biologically-active chemicals, including antioxidant polyphenols that can counteract the oxidative stress associated with obesity. This review presents polyphenols as natural complementary therapy, and a good nutritional strategy, for treating obesity without serious side effects.

scholarly journals Salvianolic Acid A Protects Against Oxidative Stress and Apoptosis Induced by Intestinal Ischemia-Reperfusion Injury Through Activation of Nrf2/HO-1 Pathways

2018 ◽  
Vol 49 (6) ◽  
pp. 2320-2332 ◽  
Author(s):  
Guo Zu ◽  
Tingting Zhou ◽  
Ningwei Che ◽  
Xiangwen Zhang
Keyword(s):  
Reactive Oxygen Species ◽  
Glutathione Peroxidase ◽  
Ischemia Reperfusion ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Salvianolic Acid ◽  
Salvianolic Acid A ◽  
Tnf Α

Background/Aims: Ischemia-reperfusion (I/R) adversely affects the intestinal mucosa. The major mechanisms of I/R are the generation of reactive oxygen species (ROS) and apoptosis. Salvianolic acid A (SalA) is suggested to be an effective antioxidative and antiapoptotic agent in numerous pathological injuries. The present study investigated the protective role of SalA in I/R of the intestine. Methods: Adult male Sprague-Dawley rats were subjected to intestinal I/R injury in vivo. In vitro experiments were performed in IEC-6 cells subjected to hypoxia/ reoxygenation (H/R) stimulation to simulate intestinal I/R. TNF-α, IL-1β, and IL-6 levels were measured using enzyme-linked immunosorbent assay. Malondialdehyde and myeloperoxidase and glutathione peroxidase levels were measured using biochemical analysis. Apoptosis was measured by terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling staining or flow cytometry in vivo and in vitro. The level of reactive oxygen species (ROS) was measured by dichlorodihydrofluorescin diacetate (DCFH-DA) staining. Western blotting was performed to determine the expression of heme oxygenase-1 (HO-1), Nrf2 and proteins associated with apoptosis. The mRNA expressions of Nrf2 and HO-1 were detected by quantitative real-time polymerase chain reaction in vivo and in vitro. Results: Malondialdehyde level and myeloperoxidase and glutathione peroxidase, TNF-α, IL-1β, and IL-6 levels group in intestinal tissue decreased significantly in the SalA pretreatment groups compared to the I/R group. SalA markedly abolished intestinal injury compared to the I/R group. SalA significantly attenuated apoptosis and increased Nrf2/HO-1 expression in vivo and in vitro. However, Nrf2 siRNA treatment partially abrogated the above mentioned effects of SalA in H/R-induced ROS and apoptosis in IEC-6 cells. Conclusion: The present study demonstrated that SalA ameliorated oxidation, inhibited the release of pro-inflammatory cytokines and alleviated apoptosis in I/R-induced injury and that these protective effects may partially occur via regulation of the Nrf2/ HO-1 pathways.

scholarly journals Selection of theIn VitroCulture Media Influences mRNA Expression of Hedgehog Genes,Il-6,and Important Genes regarding Reactive Oxygen Species in Single Murine Preimplantation Embryos

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
N. Pfeifer ◽  
D. M. Baston-Büst ◽  
J. Hirchenhain ◽  
U. Friebe-Hoffmann ◽  
D. T. Rein ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Mrna Expression ◽  
Culture Media ◽  
Expression Profiles ◽  
Reactive Oxygen ◽  
Female Reproductive Tract ◽  
Preimplantation Embryos ◽  
Oxygen Species

Background. The aim of this paper was to determine the influence of differentin vitroculture media on mRNA expression of Hedgehog genes,il-6,and important genes regarding reactive oxygen species in single mouse embryos.Methods. Reverse transcription of single embryos either culturedin vitrofrom day 0.5 until 3.5 (COOK’s Cleavage medium or Vitrolife’s G-1 PLUS medium) orin vivountil day 3.5post coitum. PCR was carried out forβ-actinfollowed by nested-PCR forshh, ihh, il-6, nox, gpx4, gpx1,andprdx2.Results. The number of murine blastocysts cultured in COOK medium which expressedil-6, gpx4, gpx1,andprdx2mRNA differed significantly compared to thein vivogroup. Except fornox, the mRNA profile of the Vitrolife media group embryos varied significantly from thein vivoones regarding the number of blastocysts expressing the mRNA ofshh, ihh, il-6, gpx4, gpx1andprdx2.Conclusions. The present study shows that differentin vitroculture media lead to different mRNA expression profiles during early development. Even the newly developedin vitroculture media are not able to mimic the female reproductive tract. The question of long-term consequences for children due to assisted reproduction techniques needs to be addressed in larger studies.

scholarly journals Reactive oxygen species and nitric oxide imbalances lead to in vivo and in vitro arrhythmogenic phenotype in acute phase of experimental Chagas disease

2020 ◽  
Vol 16 (3) ◽  
pp. e1008379 ◽  
Author(s):  
Artur Santos-Miranda ◽  
Julliane Vasconcelos Joviano-Santos ◽  
Grazielle Alves Ribeiro ◽  
Ana Flávia M. Botelho ◽  
Peter Rocha ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Reactive Oxygen Species ◽  
Chagas Disease ◽  
Acute Phase ◽  
Reactive Oxygen ◽  
Oxygen Species

scholarly journals BRAF Mutant Melanoma Adjusts to BRAF/MEK Inhibitors via Dependence on Increased Antioxidant SOD2 and Increased Reactive Oxygen Species Levels

Cancers ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 1661 ◽  
Author(s):  
Long Yuan ◽  
Rosalin Mishra ◽  
Hima Patel ◽  
Samar Alanazi ◽  
Xin Wei ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Braf Inhibitor ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Braf Mutations ◽  
Mek Inhibitors ◽  
Braf Mutant ◽  
Resistant Cells

B-Rapidly Accelerated Fibrosarcoma (BRAF) mutations are found in about 50% of melanoma patients. Treatment with Food and Drug Administration (FDA)-approved BRAF and MAP/ERK kinase (MEK) inhibitors has improved progression free and overall survival of patients with BRAF mutant melanoma. However, all responders develop resistance typically within 1 year of treatment with these inhibitors. Evidence indicates that reactive oxygen species (ROS) levels are elevated after BRAF pathway inhibition treatment. We aim to decipher the role of mitochondrial antioxidant proteins relative to ROS levels and BRAF pathway inhibitor resistance. We observed BRAF mutant melanoma cells treated with the combination of a MEK inhibitor (trametinib) and a BRAF inhibitor (dabrafenib), exhibited elevated ROS levels, both in in vitro and in vivo melanoma models. We next generated trametinib- and dabrafenib-resistant (TDR) cells and found increased ROS levels after acquisition of resistance. An immunofluorescence experiment showed an increase of DNA damage in TDR cell lines. Furthermore, we observed that TDR cells increased superoxide dismutase 2 (SOD2), an antioxidant, at both mRNA and protein levels, with the upregulation of the transcription factor Nuclear Factor (NF)-κB. Knockdown of SOD2 significantly reduced the growth of BRAF pathway inhibitor-resistant cells. In addition, the results indicate that TDR cells can be re-sensitized to BRAF pathway inhibitors by the ROS scavenger, N-Acetyl Cysteine (NAC). Overall, these data indicate that BRAF pathway inhibitor-resistant cells can compensate for elevated ROS via increased expression of the antioxidant SOD2.

Effects of D-Glucose on Chemokinesis and Resting Production of Reactive Oxygen Species in Neutrophil Granulocytes of Lean or Obese-Hyperglycemic Mouse

1997 ◽  
Vol 17 (5) ◽  
pp. 487-498 ◽  
Author(s):  
Per-Arne Oldenborg ◽  
Janove Sehlin
Keyword(s):  
Reactive Oxygen Species ◽  
Reactive Oxygen ◽  
Neutrophil Function ◽  
Neutrophil Granulocytes ◽  
Oxygen Species ◽  
Enhanced Chemiluminescence ◽  
Chronic Hyperglycemia ◽  
Locomotive Activity

The response to D-glucose (0–21 mM) was studied in neutrophil granulocytes from obese, hyperglycemic and hyperinsulinemic Umeå ob/ob mice and their lean, littermate controls in order to further elucidate the effects of in vivo and in vitro hyperglycemia on neutrophil function. Neutrophil random locomotion on glass and neutrophil resting luminol-enhanced chemiluminescence in cell suspension were studied. Random locomotion was stimulated by D-glucose in neutrophils from both Umeå ob/ob and control mice but the locomotive activity in Umeå ob/ob mouse neutrophils was significantly higher than that found in the controls at 4–21 mM glucose. In both types of mice, the stimulatory effect of D-glucose on random locomotion was diminished at 21 mM glucose (not significantly different from that at 0 mM glucose). Resting chemiluminescence from mouse neutrophils was also stimulated by glucose but here the magnitude of response was similar in neutrophils from both types of mice. These results indicate that chronic hyperglycemia and hyperinsulinemia in the Umeå ob/ob mouse may be associated with an increased neutrophil random locomotive activity but a similar resting production of reactive oxygen species, as compared with neutrophils from control mice at physiological and hyperglycemic glucose concentrations in vitro.

Sign in / Sign up

Export Citation Format

Share Document