The effects of chronic administration of cisplatin on oxidative stress in the isolated rat heart [Efekti hronične primene cisplatine na oksidacioni stres izolovanog srca pacova]

Taken into consideration that molecular and cellular mechanisms involved in cardiotoxicity are still not clear the aim of this study was to compare the production of oxidative stress parameters in the isolated rat heart between animals chronically treated with cisplatin and saline. The hearts of male Wistar albino rats (n = 24, 12 per group, age 8 weeks, body mass 250±50 g) were excised and perfused according to the Langendorff technique at gradually increased coronary perfusion pressures (40-120 cmH2O). We followed the production of superoxide anion radicals, hydrogen peroxide, and nitrites and also index of lipid peroxidation during the changes of coronary perfusion pressure (CPP) (from 40 to 120 cm H2O) in coronary venous effluent. Modifications CPP were performed in order to determined if oxidative stress is involved in coronary endothelium response in conditions of hypoxia (lower than 60 cm H2O) and hyperoxia (higher than 80 cm H2O). Based on the results of this research we can conclude that with enhancement of CPP the values of oxidative stress statistically increased. However, this increment is more prominent in control group as a result of preserved endothelium and its more powerful response to hyperoxia. On the other hand, damaged endothelium of cisplatin-treated animals had weaker response to hyperoxia, and also lower antioxidant capacity. © 2018, University of Kragujevac, Faculty of Science. All rights reserved

Peroxynitrite decomposition catalyst ameliorates renal damage and protein nitration in cisplatin-induced nephrotoxicity in rats

BACKGROUND: Oxidative stress is involved in cisplatin-nephrotoxicity. However, it has not completely established if reactive nitrogen species and nitrosative stress are involved in this experimental model. The purpose of this work was to study the role of peroxynitrite, a reactive nitrogen specie, in cisplatin-nephrotoxicity using the compound 5,10,15,20-tetrakis (4-sulfonatophenyl) porphyrinato iron (III) (FeTPPS), a soluble complex able to metabolize peroxynitrite. RESULTS: In rats treated with cisplatin (a single intraperitoneal dose of 7.5 mg/kg body weight), renal nitrosative stress was made evident by the increase in 3-nitrotyrosine on day 3. In addition, cisplatin-induced nephrotoxicity was evident by the histological damage of proximal tubular cells and by the increase in (a) serum creatinine, (b) blood urea nitrogen, and (c) urinary excretion of N-acetyl-β-D-glucosaminidase and total protein. Cisplatin-induced nitrosative stress and nephrotoxicity were attenuated by FeTPPS-treatment (15 mg/kg body weight, intraperitoneally, every 12 hours for 3 days). CONCLUSIONS: Nitrosative stress is involved in cisplatin-induced nephrotoxicity in rats. Our data suggest that peroxynitrite is involved, at least in part, in cisplatin-induced nephrotoxicity and protein nitration

Determinación de la localización sub-celular de la proteína Bik como respuesta al tratamiento con un agente antineoplásico y con un agente oxidante en dos líneas celulares de cáncer de mama

El cáncer de mama es el cáncer más frecuente en mujeres de acuerdo a la OMS (GLOBOCAN, 2012) y para combatirlo la quimioterapia es la terapia más usada a pesar de sus efectos secundarios. En particular, uno de los propósitos al usar quimioterapia es la inducción de apoptosis. La apoptosis generada por la vía mitocondrial es regulada principalmente por la familia de proteínas BCL-2. La mayoría de las cuales se localizan en el retículo endoplásmico o la mitocondria. Recientemente, algunas proteínas de esta familia han sido localizadas en sitios diferentes realizando funciones diferentes a las canónicas, sin embargo muchas de estas funciones permanecen aún sin comprenderse. BIK, un miembro pro-apoptótico de esta familia se ha encontrado desregulado en cáncer lo que ha causado gran interés en los últimos años y podría ser susceptible al estrés oxidante. Debido a que la apoptosis se asocia con la generación de estrés oxidante, el objetivo de este trabajo fue determinar si la localización subcelular de la proteína BIK se ve afectada tras un insulto oxidante inducido directamente por el tratamiento con H2O2 o indirectamente por el cisplatino el cual es un agente antineoplásico. Para ello, se realizaron experimentos en dos líneas celulares transformadas de cáncer de mama, MCF-7 y MDA-MB-231 y una línea celular epitelial de mama, MCF-10A. Los resultados mostraron que en la línea celular MCF-7, BIK se localiza dentro del citosol y después del tratamiento para inducir estrés oxidante la proteína se transloca al interior del núcleo. Sin embargo, en la línea celular MDA-MB-231, BIK se localiza en el interior del núcleo y posteriormente se transloca al citosol. De manera interesante, en la línea celular MCF-10A no hay cambio en la localización de dicha proteína después de los tratamientos, además esta línea celular mostró ser más resistente al cisplatino que las líneas celulares transformadas. Con estos resultados se concluye que la proteína BIK se localiza en diferentes compartimentos celulares dependiendo del estadio de cáncer y ésta tiene la capacidad de cambiar su localización subcelular en respuesta al estrés oxidante. Ello se asocia con una mayor sensibilidad ante la exposición a agentes tóxicos, creando así nuevas oportunidades para estudiar nuevas blancos terapéuticos que permitan el desarrollo de agentes más activos y menos dañinos

Povezanost polimorfizama NRF2, SOD2 I GPX1 gena sa pokazateljima oksidativnog distresa i prognozom kod bolesnika sa terminalnom bubrežnom slabošću

Background: Impaired redox homeostasis is a hallmark of end-stage renal disease (ESRD). Both excessive production of reactive oxygen species (ROS) and impaired antioxidant function play role in systemic oxidative stress in these patients. Polymorphisms of antioxidant genes may influence individual susceptibility towards ESRD, oxidative stress, cardiovascular complications, as well as prognosis in ESRD patients. Aims: The aim of this study was to assess the association of antioxidant nuclear factor E2-related factor 2 (Nrf2) rs6721961, superoxide dismutase 2 (SOD2) rs4880 and glutathione peroxidase 1 (GPX1) rs1050450 gene polymorphisms with the risk of ESRD development and their functional significance in terms of the level of oxidative stress byproducts and soluble cellular adhesion molecules in ESRD patients. Furthermore, the predictive power of two biomarker panels in terms of the 8-year overall and cardiovascular survival in ESRD patients was evaluated. The first biomarker panel was comprised of a specific combination of Nrf2, SOD2, GPX1 and glutathione S-transferase M1 (GSTM1) genotypes. The second biomarker panel was consisted of a combination of byproducts of oxidative stress, circulating adhesion molecules and GSTM1 deletion polymorphism. A functional role of GSTM1 deletion on endothelial dysfunction in uremic milieu was explored in vitro, using human umbilical vein endothelial cells (HUVECs). Methods: Polymorphisms of Nrf2 rs6721961, SOD2 rs4880, GPX1 rs1050450 and GSTM1 genes were determined by PCR in 256 ESRD patients and 374 controls. Byproducts of oxidative stress (thiol and carbonyl groups, advanced oxidative protein products (AOPP), nitrotyrosine, malondialdehyde (MDA), malondialdehyde adducts (MDAadd), total oxidant status (TOS) and prooxidant-antioxidant balance (PAB)), were analyzed in plasma of ESRD patients spectrophotometricaly or by ELISA. Concentration of soluble cell adhesion molecules (soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1)) was determined by ELISA. In vitro part of this study was conducted on GSTM1+/+ HUVECs and HUVECs silenced for the GSTM1 (GSTM1+/-) which were treated with 30% control or uremic serum for 6 h. Oxidative stress parameters in HUVECs were analyzed as follows: total ROS by flow cytometer, MDA by ELISA and SOD and GPX activity spectrophotometrically. Expression of 105 cytokines in HUVECs was determined by Proteome Array. Expression of ICAM-1 and VCAM-1 proteins in HUVECs was assessed by Western blot...Uvod: Poremećaj redoks homeostaze predstavlja obeležje terminalne bubrežne slabosti (TBS). Prekomerno stvaranje slobodnih radikala i smanjena antioksidativna zaštita imaju značajnu ulogu u pojavi sistemskog oksidativnog stresa kod bolesnika sa TBS. Polimorfizmi antioksidativnih gena mogu uticati na individualnu podložnost za TBS, oksidativni stres, kardiovaskularne komplikacije, kao i prognozu bolesnika sa TBS. Ciljevi: Cilj ove studije bio je da se ispita povezanost polimorfizama gena koji kodiraju regulatorne i katalitičke antioksidativne proteine, Nrf2 rs6721961, superoksid dismutazu 2 (SOD2) rs4880 i glutation peroksidazu 1 (GPX1) rs1050450 sa podložnošću za nastanak TBS, kao i njihov funcionalni značaj u pogledu nivoa pokazatelja oksidativnog oštećenja lipida i proteina i nivoa adhezionih molekula u plazmi bolesnika sa TBS. Pored toga, cilj ove studije bio je da se ispita prognostički značaj dva panela biomarkera u odnosu na opšte i kardiovaskularno preživljavanje bolesnika sa TBS nakon 8 godina praćenja. Prvi panel biomarkera sastojao se od specifične kombinacije Nrf2, SOD2, GPX1 i glutation S-transferaze M1 (GSTM1) genotipova. Drugi panel biomarkera se sastojao od kombinacije pokazatelja oksidativnog stresa, adhezionih molekula i polimorfizma GSTM1 gena. Funkcionalna uloga GSTM1 delecionog polimorfizma u nastanku endotelne disfunkcije u uremijskim uslovima ispitana je in vitro, korišćenjem kulture endotelnih ćelija (engl. human umilical vein endothelial cells - HUVECs). Materijal i metode: Polimorfizmi Nrf2 rs6721961, SOD2 rs4880, GPX1 rs1050450 i GSTM1 gena su određeni PCR metodom kod 256 TBS bolesnika i 374 pripadnika kontrolne grupe. Pokazatelji oksidativnog stresa (tiol i karbonilne grupe, napredni produkti oksidacije proteina (AOPP), nitrotirozin, malondialdehid (MDA), malondialdehid adukti (MDAadd), ukupni oksidativni status (TOS) i prooksidativni-antioksidativni balans (PAB)) analizirani su u plazmi TBS bolesnika spektrofotometrijski ili ELISA metodom. Koncentracije humanog solubilnog vaskularnog adhezivnog molekula-1 (sVCAM-1) i humanog solubilnog intracelularnog adhezivnog molekula-1 (sICAM-1) analizirani su ELISA metodom. In vitro deo ove studije sproveden je na GSTM1+/+ HUVEC ćelijama i HUVEC ćelijama utišanim za GSTM1 gen (GSTM1+/-) koje su tretirane 30% kontrolnim ili uremijskim serumom tokom 6 sati. Pokazatelji oksidativnog stresa u HUVEC ćelijama su analizirani na sledeći način: protočnim citometrom – ukupne reaktivne vrste kiseonika, ELISA metodom – MDA, spektrofotmetrijski – aktivnost SOD i GPX enzima. Metodom proteoereja je određena ekspresija 105 citokina u HUVEC ćelijama. Ekspresija ICAM-1 i VCAM-1 proteina analizirana je metodom imunoblota..

Investigation of the effects of chronic administration of cisplatin and Ru(II) complexes on the isolated heart and oxidative stress of rats

Uvod: Kompleksi rutenijuma(II) su pokazali antitumorsku aktivnost, a njihova prednost u poređenju sa cisplatinom ogleda se u potencijalno nižoj toksičnosti, dobroj selektivnosti i inhibiciji metastaza. Cilj: Cilj ove studije bio je da se ispitaju efekti kompleksa rutenijuma, [Ru(Cltpy)(en)Cl][Cl], [Ru(Cl-tpy)(dach)Cl][Cl] i [Ru(Cl-tpy)(bpy)Cl][Cl], na srce i uporede sa cisplatinom. Posebna pažnja je bila usmerena i ka ispitivanju efekata cisplatine i kompleksa rutenijuma na indukciju oksidacionog stresa. Materijal i metode: U studiju je bilo uključeno 60 W. albino pacova podeljenih u 5 grupa. Životinje su 4 nedelje primale 4 mg/kg/nedeljno komplekse rutenijuma, i cisplatinu u istoj dozi i fiziološki rastvor (4 ml/kg). Na kraju tretmana, životinje su žrtvovane, a srca su perfundovana prema Langendorff-ovom modelu. U krvi se određivao nivo markera oksidacionog stresa i funkcije srca, bubrega i jetre. Rezultati: [Ru(Cl-tpy)(en)Cl][Cl] je ispoljio negativno inotropno i hipotenzivno dejstvo i indukovao oksidacioni stres, slično kao i cisplatina, dok su histopatološka oštećenja organa bila blaža. [Ru(Cl-tpy)(dach)Cl][Cl] je ostvario negativno inotropno i hipotenzivno dejstvo i snažan oksidacioni stres, i ova dejstva su bila slabija od cisplatine. [Ru(Cl-tpy)(bpy)Cl][Cl] je ostvario slične efekte na srce kao i [Ru(Cl-tpy)(dach)Cl][Cl] i indukovao umereni oksidacioni stres. Zaključak: Profil toksičnosti [Ru(Cl-tpy)(en)Cl][Cl] smanjuje izglede za potencijalnu terapijsku primenu, dok bi [Ru(Cl-tpy)(dach)Cl][Cl] i [Ru(Cltpy)(bpy)Cl][Cl] mogli biti potencijalni kandidati za dalji razvoj, uz hemijske modifikacije koje bi umanjile toksičnost, a povećale biološku aktivnost.Introduction: Ruthenium(II) complexes have shown antitumor activity and their advantage compared to cisplatin is reflected in potentially lower toxicity, good selectivity and inhibition of metastases. Aim: The aim of this study was to examine the effects of ruthenium complexes, [Ru(Cltpy)(en)Cl][Cl], [Ru(Cl-tpy)(dach)Cl][Cl] and [Ru(Cl-tpy)(bpy)Cl][Cl], on heart and compared with cisplatin. Special attention was aimed at examining the effects of cisplatin and ruthenium complexes on induction of oxidative stress. Material and methods: 60 W. albino rats divided into 5 groups were included in study. Animals received 4 mg/kg/week of ruthenium complexes, and cisplatin in same dose and physiological solution (4 ml/kg) for 4 weeks. At the end of treatment animals were sacrificed and hearts were perfused according to Langendorff model. The level of markers of oxidative stress and heart, kidney and liver function was assessed in blood. Results: [Ru(Cl-tpy)(en)Cl][Cl] exerted negative inotropic and hypotensive effects and induced oxidative stress, similar to cisplatin, while histopathological organ damage was milder. [Ru(Cl-tpy)(dach)Cl][Cl] exerted negative inotropic and hypotensive effects and strong oxidative stress, and these effects were weaker than cisplatin. [Ru(Cl-tpy)(bpy)Cl][Cl] exerted similar cardiac effects as [Ru(Cl-tpy)(dach)Cl][Cl] and induced moderate oxidative stress. Conclusion: The toxicity profile of [Ru(Cl-tpy)(en)Cl][Cl] reduces the prospects for potential therapeutic use, while [Ru(Cl-tpy)(dach)Cl][Cl] and [Ru(Cl-tpy)(bpy)Cl][Cl] could be potential candidates for further development, with chemical modifications that would reduce toxicity and increase biological activity