In vitro effects of neuropeptide Y on migratory potential and oxidative status of human choriocarcinoma cells

Rezime: Trofoblasti su ćelije placente i karakterišu se visokom sposobnošću proliferacije, migracije i invazije. Njihova konrolisana migracija i invazija u endometrijum, ključni su procesi u implantaciji, kao i u formiranju i razvoju placente. Sposobnost trofoblasta da izvrši migraciju i invaziju na maternalno tkivo leži u osnovi pravilnog odvijanja trudnoće i pravilnog razvoja fetusa. Redukcioni poremećaji trofoblastne migracije uzrokuju aberantnu imlantaciju i plitku placentaciju, što često vodi različitim komplikacijama trudnoće koje su u vezi sa nedovoljnom placentacijom, kao što su preeklampsija i intrauterini zastoj u rastu. U nekoliko patoloških formi trudnoće pokazano je prisustvo značajno povišene koncentracije neuropeptida Y u poređenju sa koncentracijama u zdravoj trudnoći, pa je najveći deo studije fokusiran na ispitivanje potencijalnih patoloških efekata na trofoblaste. Imajući u vidu da je neuropeptid Y prisutan u određenoj koncentraciji i u plazmi zdravih trudnica, uzeti su u razmatranje i efekti ove koncentracije, putem koje je moguće da neuropeptid Y ostvari značajan fiziološki doprinos u optimalnom metabolizmu trofoblasta tokom zdrave trudnoće. Ciljevi ove studije predstavljaju in vitro ispitivanja efekata neuropeptida Y na različite aspekte funkcionisanja humanih trofoblasta. Tokom studije korišćena je ćelijska linija JEG-3 humanih trofoblasta izolovanih iz karcinoma placente, koja je pogodan model sistem za proučavanja procesa migracije i invazije. U eksperimentima su ispitivani efekti neuropeptida Y apliciranog u dve koncentracije (fiziološkoj i patološkoj) u dva vremenska tretmana (24 sata i 72 sata) sa ciljem upoređivanja njegovih fizioloških efekata i efekata koje ostvaruje u koncentracijama koje se javljaju u nekim patološkim stanjima trudnoće. Poseban fokus studije je bio na utvrđivanju potencijalnog doprinosa ovog molekula na promene migratorne i invazivne sposobnosti trofoblasta u njegovim koncentracijama višim od fizioloških. Koncentracije NPY od 0,1 nmol/L i 1 nmol/L korišćene su za testiranje efekata na apoptotski indeks (preživljavanje ćelija), vijabilnost, migracioni kapacitet, oksidativni / antioksidativni status, produkciju metaloproteinaza (MMR-9), kao i za merenje ekspresije gena (iNOS, HIF-1α, VEGF, COX-2 i MMP-9) i distribucije nekih proteina u ćeliji (iNOS, E-kadherin). Kontrolne ćelije su gajene u čistom medijumu za kultivaciju da bi se dobile bazalne vrednosti ispitivanih parametara. Eksperimentalni podaci dobijeni tokom ove studije pokazali su da neuropeptid Y apliciran u fiziološkoj (0,1 nmol/L) i patološkoj (1 nmol/ L) koncentraciji ostvaruje različite efekte na parametre oksidativnog / antioksidativnog metabolizma, migracije, kao i na vijabilnost, apoptotoski indeks i proliferativnu sposobnost humanih trofoblasta linije JEG-3. Najznačajniji efekat fiziološke koncentracije neuropeptida Y je povećavanje vijabilnosti trofoblasta, što ukazuje na značajan doprinos održavanju nivoa oksidtivnog metabolizma u trofoblasima koji prirodno funkcionišu u stanju hipoksije. Fiziološka koncentracija neuropeptida Y ne indukuje značajan oksidativni stres, ukazujući da trofoblasti pri ovoj koncentraciji efikasno mogu neutralisati produkte povišene aktivnosti mitohondrijalnog lanca respiracije. Neuroptpid Y apliciran u koncentraciji od 0,1 nmol/L pokazuje protektivan efekat na preživljavanje trofoblasta, kao i značajan proliferativan efekat. Ovi podaci ukazuju da NPY u fiziološkim koncentracijama može imati doprinos u ostvarivanju funkcija trofoblasta neophodnih za optimalnu implantaciju i placentaciju.Abstract: Trophoblasts cells are placental cell type characterized by high proliferation capacity, migration and invasion. Highly regulated trophoblast migration and invasion of the endometrium, are the essentail process in the implantation and development of the placenta. The migration and invasion of trophoblast cells of maternal tissues are basis of a proper progress of pregnancy and optimal fetal development. Reduced trophoblast migration and invasion cause abberant implantation and shallow placentation, often leading to a various complications of pregnancy that are related with inadequate placentation, such as preeclampsia and intrauterine growth restriction. In several pathological forms of pregnancy the presence of significantly elevated concentration of neuropeptide Y in comparison with its concentrations in healthy pregnancy was recorded, so the largest part of the study were focused on the examination of potential pathological effects of NPY on trophoblast cells. Since neuropeptide Y is also present in a particular concentration in plasma of women with healthy pregnancy, the effects of this physiological concentration by which neuropeptide Y is possible to significantly contribute to the optimal physiological metabolism of trophoblasts during a healthy pregnancy were taken into consideration. The objectives of this study were to investigate in vitro effects of neuropeptide Y on different aspects of the functioning of the human trophoblast. During the study we used the cell line JEG-3 of human trophoblast, isolated from placental cancer, which represents a suitable model system for examinating of the processes of cell migration and invasion. In the experiments we have investigated the effects of neuropeptide Y applied in two concentrations (physiological and pathological) in two different time treatment (24 and 72 hours) with the aim of comparing its physiological effects and the effects achieved by its pathological concentrations present in some pregnancy disorders. A particular focus of the study was to determine the potential contribution of this molecule on changes of the migratory and invasive ability of trophoblast cells induced by concentrations higher than physiological. NPY concentration of 0,1 nmol/L and 1 nmol/L were used to examine the effects on apoptotic index (survival rate of the cells), cell viability, migration capacity, the oxidative / antioxidative status, the production of metalloproteinase (MMP-9), gene expression (iNOS, HIF-1α, VEGF, COX-2 and MMP-9) and the distribution of some proteins in cell (iNOS, E-cadherin). Control cells were grown in pure culture mediim to obtain basal values of the examined parameters. Experimental data obtained in this study showed that neuropeptide Y administered in physiological (0,1 nmol/L) and pathological (1 nmol/L) concentration achieves various effects on the parameters of the oxidative / antioxidative metabolism, migration and the viability index apoptotic index and proliferative rate of human trophoblast cell line JEG -3. The most significant effect of physiological concentrations of neuropeptide Y is increasing trophoblast viability, which indicates a significant contribution to maintaining the levels of oxidative metabolism trofoblasts, naturally operating in a state of hypoxia. Physiological concentrations of neuropeptide Y does not induce significant oxidative stress, indicating that trophoblasts at this concentration can effectively neutralize the products of elevated activity of mitochondrial respiratory chain. Neuropetpide Y administered in a concentration of 0,1 nmol/L also showed protective effect on the survival of the trophoblast, as well as a significant proliferative effect. These data suggest that NPY in physiological concentrations may have a contribution in achieving the function of a trophoblast necessary for optimal implantation and placentation

Paracetamol-induced changes of haemato-biochemical and oxidative stress parameters in rat blood: protective role of vitamin C and betaglucan

Paracetamol (acetaminophen) is widely used as an ov er-the-counter analgesic and antipyretic drug. The aim of this stu dy was to investigate the possible protective effects of vitamin C (100mg/kg/day i.p.) and β -glucan (40 mg/kg/day i.p.) on altered haematological, biochemical and oxidative s tress parameters in the blood of rats treated with paracetamol (100 mg/kg/day i.p.) for 3 days. Exposure of rats to paracetamol caused changes of some haematological parameters (R BCs count, Hb concentration, Ht value and WBCs count), suggesting that the paraceta mol induced haematotoxicity. Paracetamol reduced serum total protein (TP), album in and globulin, while increased alanine aminotransferase (ALT), aspartate aminotran sferase (AST) and lactate dehydrogenase (LDH) activities compared to the cont rol. The results indicate that paracetamol is led to significant decrease in the c oncentration of Na + and K + and increase of Ca 2+ in the serum compared to the control. Coadministra tion of vitamin C and β -glucan with paracetamol reversed these changes of haematol ogical and biochemical parameters and diminished the toxic effects of paracetamol. Th e obtained results indicated that the concentration of LPO in erythrocytes significantly increased in, while the concentration of GSH significantly decreased in the group treated with paracetamol compared to control group. Coadministration of vitamin C and β -glucan with paracetamol reversed paracetamol-induced alterations in these oxidative stress parameters. This study suggests that paracetamol has significant prooxidative effec ts and may disrupt oxidant/antioxidant balance in erythrocytes. Furthermore, coadministrat ion with vitamin C and β -glucan have protective effects on paracetamol-induced oxidative damage and haematotoxicity.Kragujevac Journal of Science (2016), 38: 135-14

Lipid Peroxidative Damage on Cisplatin Exposure and Alterations in Antioxidant Defense System in Rat Kidneys: A Possible Protective Effect of Selenium

Cisplatin (Cis-diamminedichloroplatinum II, CP) is an important chemotherapeutic agent, useful in the treatment of several cancers, but with several side effects such as nephrotoxicity. The present study investigated the possible protective effect of selenium (Se) against CP-induced oxidative stress in the rat kidneys. Male Wistar albino rats were injected with a single dose of cisplatin (7 mg CP/kg b.m., i.p.) and selenium (6 mg Se/kg b.m, as Na2SeO3, i.p.), alone or in combination. The obtained results showed that CP increased lipid peroxidation (LPO) and decreased reduced glutathione (GSH) concentrations, suggesting the CP-induced oxidative stress, while Se treatment reversed this change to control values. Acute intoxication of rats with CP was followed by statistically significant decreased activity of antioxidant defense enzymes: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and glutathione-S-transferase (GST). Treatment with Se reversed CP-induced alterations of antioxidant defense enzyme activities and significantly prevented the CP-induced kidney damage

Zaštitno djelovanje kvercetina i vitamina C protiv nikotinom izazvane toksičnosti u krvi Wistar štakora

Nicotine is a potential inducer of oxidative stress, through which it can damage numerous biological molecules. The aim of our study was to investigate the prooxidative effects of nicotine and protective (additive or synergistic) effects of quercetin and vitamin C in the blood of experimental animals, to determine whether the combination of these antioxidants might be beneficial for clinical purposes. Wistar albino rats were receiving intraperitoneal nicotine injection (0.75 mg kg-1 per day) or saline (control group) or nicotine plus quercetin (40 mg kg-1 per day) and vitamin C (100 mg kg-1 per day) for three consecutive days. On day 4, we determined their blood lipid profile, liver enzymes, oxidative stress parameters, and antioxidative system parameters. Compared to untreated control, nicotine significantly increased total cholesterol, LDL-cholesterol, triglycerides, liver enzymes (alanine transaminase, aspartate transaminase, and lactate dehydrogenase) and oxidative stress parameters (superoxide anion, hydrogen peroxide, and lipid peroxide) and decreased HDL-cholesterol, glutathione, and superoxide dismutase/catalase activity. Quercetin + vitamin C reversed these values significantly compared to the nicotine alone group. Our results confirm that nicotine has significant prooxidative effects that may disrupt the redox balance and show that the quercetin + vitamin C combination supports antioxidant defence mechanisms with strong haematoprotective activity against nicotine-induced toxicity. In practical terms, this means that a diet rich in vitamin C and quercetin could prevent nicotine-induced toxicity and could also be useful in the supportive care of people exposed to nicotine.Nikotin je potencijalni induktor oksidacijskoga stresa, preko kojega može oštetiti brojne biološke molekule. Cilj našega istraživanja bio je ispitati prooksidacijsko djelovanje nikotina i zaštitno (aditivno ili sinergističko) djelovanje kvercetina i vitamina C u krvi eksperimentalnih životinja te utvrditi može li kombinacija tih antioksidansa biti korisna u kliničke svrhe. Wistar albino štakori primali su intraperitonealno injekciju nikotina (0,75 mg kg-1 po danu) ili fiziološke otopine (kontrolna skupina) ili nikotina s kvercetinom (40 mg kg-1 po danu) i vitaminom C (100 mg kg-1 po danu) tri uzastopna dana. Četvrtoga dana odredili smo lipidni profil u krvi, jetrene enzime, parametre oksidacijskoga stresa i antioksidacijskoga sustava. U usporedbi s netretiranom kontrolnom skupinom, nikotin je značajno povećao ukupni kolesterol, LDL-kolesterol, trigliceride, jetrene enzime (alanin transaminaze, aspartat transaminaze i laktat dehidrogenaze) i parametre oksidacijskoga stresa (superoksid anion, vodikov peroksid i lipidne perokside), a smanjio HDL-kolesterol, glutation i aktivnosti superoksid dismutaze/katalaze. Kvercetin i vitamin C značajno su utjecali na te vrijednosti u odnosu na skupinu samo s nikotinom. Naši rezultati potvrdili su značajno prooksidacijsko djelovanje nikotina koje može poremetiti redoks ravnotežu i pokazuje da kombinacija kvercetina i vitamina C podržava antioksidacijske obrambene mehanizme s jakim hematoprotekcijskim aktivnostima protiv nikotinom izazvane toksičnosti. Možemo zaključiti da prehrana bogata kvercetinom i vitaminom C može koristiti kao prevencija nikotinom inducirane toksičnosti te da kombinacija tih dvaju antioksidansa može biti korisna u kliničkom oporavku ljudi izloženih nikotinu

Antioxidative and haematoprotective activity of coenzyme Q10 and vitamin E against cadmium-induced oxidative stress in Wistar rats.

Cadmium (Cd) is a major environmental pollutant, which exerts adverse effects mainly by inducing oxidative stress. Coenzyme Q10 (CoQ10) and vitamin E (VE), naturally occurring antioxidants, improve health condition by inactivating free radicals and enhancing antioxidative defence. The aim of our study was to investigate the protective role of CoQ10 and/or VE pretreatment against Cd-induced haematotoxicity. Wistar albino rats were intramuscularly injected with CoQ10 (20 mg/kg b.w.) and/or VE (20 IU/kg b.w.) or with saline (control group). After 24 h, Cd was injected intraperitoneally (0.4 mg/kg b.w.) and 1 day after, animals were sacrificed. Acute Cd intoxication caused significant changes in haematological and biochemical parameters and altered the glutathione cycle, leading to the formation of lipid peroxidation, while the concentrations and activities of antioxidants (vitamins C and E, superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase) were decreased. CoQ10 and/or VE significantly maintained these values to near-normal levels, afforded additional protection by reducing lipid peroxidation and improved the levels of antioxidants in the blood. Plasma CoQ10 and VE levels negatively correlated with oxidative damage parameters while positively correlated with antioxidative defence parameters. Regarding their effects, CoQ10 and VE were in synergistic interaction. The present study suggested that CoQ10 and VE combination may be beneficial in protecting from Cd-induced haematotoxicity and may be used as a preventive against acute Cd intoxication of exposed people.Toxicology and industrial health (2017), 33(10): 746-75

Role of selenium and vitamin C in mitigating oxidative stress induced by fenitrothion in rat liver.

Excessive use of organophosphate insecticides, including fenitrothion (FNT) can cause detrimental consequences in non-target organisms. Selenium (Se) and vitamin C (Vit C) possess protective abilities against various toxic compounds due to their antioxidative properties. Accordingly, the aim of the present study was to examine the possible ameliorative effects of Se and Vit C in hepatotoxicity induced by FNT. For the purpose of this study, male Wistar albino rats were divided into control and groups treated with Se (0.5 mg/kg b.w, as Na2SeO3) and Vit C (100 mg/kg b.w), FNT (20 mg/kg b.w) and FNT in cotreatment with Se and Vit C for 30 days. The current data showed a reduction in absolute and relative liver weight after FNT administration. Increased activities of liver enzymes (AST, ALT, ALP, LDH and GGT) indicated liver damage. FNT alone caused significant alterations in biochemical parameters (glucose and total bilirubin). Elevation in LPO level along with decreased activities of antioxidant enzymes (SOD, CAT, GSH-Px) and GSH content reflected the presence of oxidative stress. Coadministration of FNT with Se and Vit C exhibited hepatoprotective role confirmed by reduction of oxidative stress levels and restoration in the values of examined parameters. Because of their beneficial effects, Se and Vit C may be used in reducing injuries caused by pesticides

Hematoprotective effects and antioxidant properties of β-glucan and vitamin C against acetaminophen-induced toxicity: an experimental study in rats.

Acetaminophen is widely used as an over-the-counter analgesic and antipyretic drug. The aim of the present study was to investigate the pro-oxidative effects of acetaminophen (300 mg/kg/day i.p.) and antioxidative effects of β-glucan (4 mg/kg/day i.p.) and/or vitamin C (100 mg/kg/day i.p.) on the blood parameters of treated rats. After 3 days of treatment, hematological and parameters of redox status were measured. Exposure of rats to acetaminophen caused significant changes in some hematological parameters and the glutathione redox cycle, leading to an increased concentration of oxidative stress parameters and the formation of lipid peroxidation, while the activities of antioxidant enzymes were decreased. Administration of β-glucan and/or vitamin C reduced lipid peroxidation and restored the levels of examined hematological and oxidative stress parameters and improved the activities of antioxidant enzymes. Obtained results demonstrated that acetaminophen has significant pro-oxidative effects and may disrupt redox balance in blood of rats, while the combination of β-glucan and/or vitamin C amplified the antioxidant defense potential and exhibited a strong hematoprotective activity against acetaminophen-induced toxicity. Therefore, β-glucan and vitamin C co-treatment may be a promising therapeutic option for the treatment of acute acetaminophen hematotoxicity

Material characterization of the main steam pipeline made of 12%Cr tempered martensite ferritic steel after prolonged service

In the past decades, tempered martensitic ferritic steels with 9-12% chromium and containing carbide stabilizing elements have been widely used for components which operate within the creep range in power and industrial processing plants. The investigated samples made of X20CrMoV12-1 steel were taken from the main steam pipeline in thermal power plant after 250.000 h (545°C and 19MPa). The low values of impact toughness are the consequence of aging and weakening of grain boundaries, despite the fact that aging processes have not significant effects on the microstructural degradation and drop of the other mechanical properties

Material characterization of the main steam pipeline made of 12%Cr tempered martensite ferritic steel after prolonged service

In the past decades, tempered martensitic ferritic steels with 9-12% chromium and containing carbide stabilizing elements have been widely used for components which operate within the creep range in power and industrial processing plants. The investigated samples made of X20CrMoV12-1 steel were taken from the main steam pipeline in thermal power plant after 250.000 h (545°C and 19MPa). The low values of impact toughness are the consequence of aging and weakening of grain boundaries, despite the fact that aging processes have not significant effects on the microstructural degradation and drop of the other mechanical properties

The ameliorating effects of selenium and vitamin C against fenitrothion-induced blood toxicity in Wistar rats

Fenitrothion is widely used organophosphate pesticide in agriculture and health programs, but besides, it causes several toxic effects. The present study was designed to evaluate the possible protective effects of selenium (0.5 mg/kg b.w.) and vitamin C (100 mg/kg b.w) on altered haematological, biochemical and oxidative stress parameters in the blood of rats orally treated with fenitrothion (20 mg/kg b.w) for 30 days. Fenitrothion caused changes in body weight, food and water intake, and some haematological and biochemical parameters. Fenitrothion altered the glutathione redox status (GSH and GSSG) and decreased activity of antioxidant enzymes (GSH-Px, GST, SOD and CAT), leading to a lipid peroxidation. Selenium and vitamin C, by improving the activity of antioxidants, reduced oxidative stress and a lipid peroxidation, maintaining the values of examined parameters to optimal levels. Therefore, selenium and vitamin C could be useful in providing protection of exposed non-target organisms including people from fenitrothion.Environmental Toxicology and Pharmacology (2017), 56: 204-20