In Vitro Study of Antioxidant Activity of Serum and Plasma Samples As Well As Glutathione Exposed to Various Exogenous Stress Factors

Author(s):  
Małgorzata Olszowy-Tomczyk ◽  
Łukasz Paprotny ◽  
Agnieszka Celejewska ◽  
Dorota Szewczak ◽  
Dorota Wianowska

Abstract The imbalance between the production of Reactive Oxygen Species (ROS) and their sequestration promotes the formation of so-called oxidative stress conditions which are considered crucial in the aging process and development of many human diseases. Glutathione plays an essential role in the antioxidative barricade against ROS. Its role in the detoxification process of xenobiotics and carcinogen is also known. However, there are no comparative studies on the antioxidant properties of both biological samples and glutathione as well as the change in these properties as a result of exposure to various stress factors. This paper fills this gap comparing the antioxidant activity of serum and plasma samples of the known glutathione content with the activity of glutathione itself assessed by the different methods. In addition, it reveals a significant role of environmental xenobiotics in oxidative stress and differentiates the stress induced by different groups of drugs, among which the greatest one has been demonstrated for antiarrhythmic drugs and cytostatics. More importantly, it proves that human plasma is more resistant to stress factors and N-acetylcysteine clearly promotes the extension of antioxidant properties of both the plasma and serum samples. The latter conclusion is consistent with the implied preventive and/or supportive action of this drug against SARS-CoV-2.

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Zonghao Tang ◽  
Jiajie Chen ◽  
Zhenghong Zhang ◽  
Jingjing Bi ◽  
Renfeng Xu ◽  
...  

The increase of oxidative stress is one of the important characteristics of mammalian luteal regression. Previous investigations have revealed the essential role of reactive oxygen species (ROS) in luteal cell death during luteolysis, while it is unknown how ROS is regulated in this process. Considering the decrease of blood flow and increase of PGF2α during luteolysis, we hypothesized that the HIF-1α pathway may be involved in the regulation of ROS in the luteal cell of the late corpus luteum (CL). Here, by using a pseudopregnant rat model, we showed that the level of both HIF-1α and its downstream BNIP3 was increased during luteal regression. Consistently, we observed the increase of autophagy level during luteolysis, which is regulated in a Beclin1-independent manner. Comparing with early (Day 7 of pseudopregnancy) and middle CL (Day 14), the level of ROS was significantly increased in late CL, indicating the contribution of oxidative stress in luteolysis. Inhibition of HIF-1α by echinomycin (Ech), a potent HIF-1α inhibitor, ameliorated the upregulation of BNIP3 and NIX, as well as the induction of autophagy and the accumulation of ROS in luteal cells on Day 21 of pseudopregnancy. Morphologically, Ech treatment delayed the atrophy of the luteal structure at the late-luteal stage. An in vitro study indicated that inhibition of HIF-1α can also attenuate PGF2α-induced ROS and luteal cell apoptosis. Furthermore, the decrease of cell apoptosis can also be observed by ROS inhibition under PGF2α treatment. Taken together, our results indicated that HIF-1α signaling is involved in the regression of CL by modulating ROS production via orchestrating autophagy. Inhibition of HIF-1α could obviously hamper the apoptosis of luteal cells and the process of luteal regression.


2021 ◽  
Vol 2021 ◽  
pp. 1-17
Author(s):  
Shiyao Xue ◽  
Hongdong Han ◽  
Shunli Rui ◽  
Mengliu Yang ◽  
Yizhou Huang ◽  
...  

Previous studies on serum fetuin-B (fetuin-like protein IRL685) have investigated its association with T2DM; however, the reason for the variation in serum fetuin-B and its regulatory factors in metabolic disease remain unclear. Here, we evaluated serum fetuin-B levels in women with newly diagnosed MetS and performed multiple interventions to investigate the role of fetuin-B in the pathogenesis of MetS. Serum fetuin-B levels were assessed using ELISA. Bioinformatics analysis was performed to analyze fetuin-B-related genes and signaling pathways. Additionally, oxidative stress parameters were measured in the in vitro study. For subgroup analyses, we performed EHC, OGTT, and treatment with a GLP-1RA to investigate the regulatory factors of serum fetuin-B. We found that in comparison with healthy subjects, serum fetuin-B levels were markedly increased in women with MetS. Further, serum fetuin-B showed a positive correlation with WHR, FAT%, TG, FBG, HbA1c, FIns, HOMA-IR, VAI, and LAP. Bioinformatics analysis revealed that most fetuin-B-related core genes were involved in cholesterol metabolism and fat decomposition. Consistent with this finding, multivariate regression analysis showed that triglyceride content and WHR were independently associated with serum fetuin-B. We also observed that serum fetuin-B levels were markedly elevated in healthy subjects after glucose loading and in women with MetS during EHC. In vitro, overexpression of fetuin-B promoted oxidative stress in HepG2 cell. After 6 months of treatment with a GLP-1RA, serum fetuin-B levels in women with MetS decreased following an improvement in metabolism and insulin sensitivity. Therefore, serum fetuin-B is associated with MetS, which may serve as a biomarker of oxidative stress. This trial is registered with ChiCTR-OCC-11001422.


Author(s):  
Mona Pourjafar ◽  
Sara Malih ◽  
Akram Ranjbar

: In recent years, the applications of nanoparticles have received a great attention due to their industrial and biomedical applications, while their beneficial effects suffer from controversial results at clinical stages. In the current study, cytotoxicity of cerium oxide (CeNP) nanoparticles (100 nm) were evaluated using mitochondria derived from wistar rat's liver. Isolated mitochondria from rat’s liver were divided into 7 groups including group 1 as control and group 2 to 7 as treatment group with different doses of CeNP (5, 10, 50, 100, 250 and 500mg/ml, respectively), for 24,48 and 72 hours. After exposure, oxidative stress biomarkers such as total ‎antioxidant capacity (TAC), lipid peroxidation (LPO), total thiol groups (TTG), catalase activity (CAT) and mitochondrial viability, were determined in isolated rat liver mitochondria. Results have shown that CeNPs increase TAC, TTG, CAT, LPO and viability of mitochondria in various exposure times and confirm antioxidant properties of CeNPs in mithocondria while mitochondria is a main source for the generation of reactive oxygen species (ROS).


2021 ◽  
Vol 75 (1) ◽  
pp. 889-895
Author(s):  
Małgorzata Lewicka ◽  
Magdalena Zawadzka ◽  
Gabriela Henrykowska ◽  
Maciej Rutkowski ◽  
Andrzej Buczyński

Abstract Objectives The article presents the results of an in vitro study aimed at identifying changes in parameters of oxidative stress – concentration of malondialdehyde (MDA), enzymatic activity of superoxide dismutase (SOD-1) and protective antioxidant role of melatonin (MLT) during the exposure of blood platelets to electromagnetic radiation (EMR) emitted by monitors. Methods Platelets were exposed to an EMR for 30- and 60 min. generated by monitors (1 kHz frequency, 220 V/m intensity). In each sample the level of SOD-1 activity and concentration of MDA were determined. Results The MDA concentration increased significantly after 30-and 60-min. irradiation, as compared to control values (2.53 vs 1.36; 3.64 vs 1.36 nmol/109 blood platelets) and after the addition of MLT it decreased (2.53 vs 1.55; 3.64 vs 1.12 nmol/109 blood platelets). The activity of SOD-1 increased significantly compared to control values after 30 min. and 60 min. of exposure to EMR (1.97vs 0.75; 2.08 vs 0.75 U/g of protein), and significantly decreased after the addition of MLT only in samples exposed for 60 min. (2.08 vs 0.95 U/g of protein). Discussion The results demonstrated the possibly negative effect of EMR on oxygen metabolism of blood platelets and indicated a possible protective role of melatonin in this process.


2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
R Carnevale ◽  
V Cammisotto ◽  
C Nocella ◽  
S Bartimoccia ◽  
D Pastori ◽  
...  

Abstract Background High circulating levels of proprotein convertase subtilisin/kexin 9 (PCSK9) were shown to be predictive of cardiovascular events (CVEs) in patients with atrial fibrillation (AF). Because high PCSK9 plasma levels were significantly correlated with 11-dehydro-thromboxane B2 (11-dh-TxB2), a marker of platelet activation, it is conceivable to hypothesize a direct effect of PCSK9 on platelet activation but the mechanism is still unclear. Purpose We evaluated the association between PCSK9 and platelet activation in FA patients and investigate the possible molecular mechanism involved. Methods According to our previous prospective study, we conducted a post-hoc analysis including 50 patients with baseline PCSK9 below and 50 above the median value of 1200pg/ml. The two groups were balanced for age and sex. In vivo platelet activation was assessed by aggregation (PA), recruitment, plasma thromboxane B2 (TxB2) formation and sPselectin levels. As markers of oxidative stress we used sNox2-dp, H2O2 production, urinary isoprostanes and oxLDL. To asses the role of PCSK9 in platelet activation, we performed an in vitro study with platelets from healthy subjects (n=5) added with PCSK9 concentrations achievable in human circulation (1000pg/ml and 2000pg/ml) measuring PA, TxB2, isoprostanes production, Nox2 activation, H2O2 production, oxLDL, p38, p47 and PLA2 phosphorylation. Results We observed an increased of platelet activation and oxidative stress in patients with PCSK9 levels above median (1200pg/ml) compared to those below (p<0.05). A significant correlation between plasma levels of PCSK9 and markers of platelet activation and markers of oxidative stress were found. In vitro study demonstrated that PCSK9, at the concentration similar to that of patients with CVEs, was able to increase platelet activation act by binding oxLDL receptor. PCSK9 dependent platelet activation is mediated by p47 phosphorylation, a key step in Nox2 activation and is mediated by the PLA2 phosporylation. Conclusions PCSK9, at concentration achievable in patients with CVEs, increased platelet aggregation via oxLDL receptor with a pathway involving Nox2 activation.


2020 ◽  
Vol 10 (7) ◽  
pp. 2374 ◽  
Author(s):  
Rosaria Arena ◽  
Simona Manuguerra ◽  
Edward Collins ◽  
Abdelkarim Mahdhi ◽  
Giuseppe Renda ◽  
...  

The aim of this paper was to obtain different extracts from the aerial parts of Mesembryanthemum nodiflorum, comparing traditional extraction (using solvents with increasing polarity such as hexane, ethanol 80%, acetone 70% and water) with an eco-friendly technique (supercritical fluid extraction (SFE)); to evaluate which extract showed a higher amount of antioxidants and then evaluate the bioactive properties in vitro, in human fibroblast (HS68). From the six extracts obtained it was observed that the solvent with the highest extraction efficiency was water, but the extracts in ethanol, N-hexane and SFE are those that showed the highest antioxidant activity (polyphenols, 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and reducing power). On the basis of these results, the SFE extract was chosen to evaluate the antioxidant and anti-aging activity in vitro, by assessing cell vitality and molecular markers (MTT and immunoblotting assays). The results showed that the SFE extract exerted antioxidant activity in vitro, protecting cells from mortality induced by oxidative stress; this protection was also confirmed at the molecular level, by the levels of the protein integrin -1, that is able to prevent the negative effects of a stress situation, such as oxidative stress, that could promote aging and related diseases. This extract, obtained with an eco-friendly technique, given its beneficial properties, could be used for application in nutraceuticals and cosmeceuticals.


Antioxidants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 318
Author(s):  
Ida Ceravolo ◽  
Federica Mannino ◽  
Natasha Irrera ◽  
Francesco Squadrito ◽  
Domenica Altavilla ◽  
...  

Fuchs endothelial corneal dystrophy (FECD) is characterized by the gradual deterioration of corneal endothelial cells (CECs) and is the most common cause of corneal transplantation worldwide. CECs apoptosis caused by oxidative stress plays a pivotal role in the pathogenesis of FECD. Antioxidant compounds have been of considerable significance as a candidate treatment in the management of corneal diseases. Based on these findings, the objective of this study was to evaluate the effects of an aloe extract with antioxidant properties, in an “in vitro” model of FECD. Human corneal epithelial (HCE) cells were preincubated with aloe extract 100 μg/mL, two hours before hydrogen peroxide (H2O2) stimulus. H2O2 challenge significantly reduced the cell viability, increased the generation of Reactive Oxygen Species (ROS) and malondialdehyde levels. Moreover, m-RNA expression and activity of Nrf-2, Catalase and Superoxide dismutase (SOD) were reduced together with an enhanced expression of IL-1β, tumor necrosis factor-α (TNF-α), IL-6, and cyclooxygenase 2 (COX-2). Furthermore, Bcl-2, Caspase-3 and Caspase-8 expression were down-regulated while Bax was up-regulated by H2O2 stimulus. Aloe extract blunted the oxidative stress-induced inflammatory cascade triggered by H2O2 and modulated apoptosis. Aloe extract defends HCE cells from H2O2-induced injury possibly due its antioxidant and anti-inflammatory activity, indicating that eye drops containing aloe extract may be used as an adjunctive treatment for FECD.


2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
L. C. Cruz ◽  
J. E. S. Batista ◽  
A. P. P. Zemolin ◽  
M. E. M. Nunes ◽  
D. B. Lippert ◽  
...  

We characterized, for the first time, the quality and identity of Brazilian Pampa biome honey and its antioxidant propertiesin vitro(FRAP, DDPH and ABTS). The potential protective effect of honey against oxidative stress induced by iron (Fe) and paraquat, (PQ) in aDrosophila melanogastermodel (in vivo) was also tested. The results indicated that all honey samples tested showed antioxidant activityin vitro. Flies treated with honey showed increased lifespan and were protected against oxidative stress induced by Fe and PQ. Despite the high concentration of sugars in honey (approximately 70–80%), our results demonstrate a hypoglycemic-like effect of honey inDrosophila. Thus, this study demonstrates the high quality of Brazilian Pampa biome honey as well as its significant antioxidant activityin vitroandin vivo, pointing to the potential use of this natural product as an alternative in the therapy of oxidative stress-associated diseases.


Author(s):  
Dipti Acharya ◽  
Umesh Kumar Gilhotra ◽  
Ranjan Singh ◽  
Himanshu Rajpurohit ◽  
Ajay Gar

Antioxidant play an important role to protect damage caused oxidative stress (OS).Chlorophyll having phenolic contents are reported to possess antioxidant properties. The present study was designed to investigate the antioxidant properties of methanolic solution from Super Chlorophyll. Objective: The main objective of the study was to evaluate the antioxidant activity of the chlorophyll in different validated in vitro models. Methods: The antioxidant activities of methanolic solution were evaluated by in vitro standard method using UV-Spectrophotometer. The antioxidant activity were determined bt total antioxidant capacity, DPPH (1-1-diphennyl-2-picrylhydrzine) radical scavenging assay, Nitric Oxide scavenging assay and Hydrogen Peroxide scavenging assay methods. Result: The solution of Chlorophyll was studied for antioxidant potential. Ascorbic acid was used as standard. This method is simple and activity of the solution is reported in term of IC50 value. Antioxidant properties depend on the IC50 value. Lesser the IC50 value, more is the antioxidant activity. Chlorophyll showed good antioxidant activity. Conclusion:  In case of in vitro antioxidant activity, Chlorophyll scavenged DPPH, hydrogen peroxide, nitric oxide radicals significantly, showed IC50 values near to standard ascorbic acid, thus proving to have good antioxidant potential. Keywords: Oxidative stress, DPPH, Antioxidant, Super chlorophyll.


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