Effect of bovine CuZn superoxide dismutase on C3 clone of B-16 mouse melanome cells in culture

B-I6/C3 mouse melanoma cells undergo melanogenesis and differentiation 9 days after plating under usual conditions. In our experiments the effect of exogenous bovine CuZn superoxide dismutase (CuZn SOD) on B-I6/C3 cells in cultures was studied. The exogenous CuZn SOD was added, 24 hours after cell plating, in growth medium, which either contain or not Fetal Calf Serum (FCS). B-I6/C3 melanoma cells growth was followed over 5 days. Different effect of CuZn SOD on the culture was observed in relation to FCS present in growth medium. CuZn SOD induced a dose dependent increase in melanin content of B-I6/C3 cells in serum deprived medium. Our results are discussed in respect to dismutating CuZn SOD activity, which may act to an enhanced level of oxidative stress, related to the higher metabolism in melanoma cells. On the other hand we supposed that CuZn SOD produced an elevated level of hydrogen peroxide. Therefore hydrogen peroxide than may play some role in cell differentiation as transmembrane messenger.Ćelije B-I6/C3 melanoma miša melaniziraju i diferenciraju devetog dana posle zasađivanja pod uobičajenim uslovima. U našim eksperimentima smo proučavali uticaj egzogeno dodavane CuZn superoksid-dismutaze ( CuZn SOD) na ćelije B-I6/C3 u kulturi. Egzogena CuZn SOD je dodavana 24 časa posle zasađivanja ćelija u medijume za rast koji su sadržavali ili ne goveđi fetalni serum. Rast ćelija B-I6/C3 je praćen pet dana. Zapažen je različit uticaj CuZn SOD na ćelijske kulture gajene sa serumom ili bez njega. CuZn SOD izaziva dozno zavisnu sintezu i nakupljanje melanina u ćelijama koje su rasle u medijumu koji nije sadržavao goveđi fetalni serum. Naši rezultatu se mogu diskutovati u svetlu dismutacione aktivnosti CuZn SOD koja deluje na povećani nivo oksidacionog stresa izazvanog visokim metabolizmom melanoma ćelija. S druge strane, pretpostavili smo da CuZn SOD svojom aktivnošću proizvodi više vodonik peroksida. Vodonik peroksid, nadalje, može da ima izvesnu ulogu u diferencijaciji ćelija kao transmembranski signalni molekul.Projekat ministarstva br. 166

Aktivnost superoksid-dismutaze tokom maturacije kulture ćelija b-16 melanoma miša

The C3 clone of B-16 mouse melanoma was cultured for 1, 6 and 9 days and analyzed. The changes which are not directly linked to melanogenesis in the B-16 / C3 cultures during their maturation were characterized. Early (1 day) confluent (6 days) and old (9 days) cell cultures are distinguished by their leucine aminopeptidase (LAP) and α-naphthyl acetate esterase (ANAE) isoenzyme patterns. Both quantitative and qualitative changes in LAP and ANAE isoenzyme can be observed during culture maturation. There is an increase in the activity of the enzyme copper, zinc-containing superoxide-dismutase (CuZn SOD). The increaase in the CuZn SOD enzyme activity might be related to B-16/C3 cell melanogenesis and / or to differentiation.C3 klonovi B-16 melanoma miša su gajeni u kulturi 1, 6 i 9 dana i potom analizirani. Izvršena je karakterizacija promena B-16/C3 ćelijskih kultura tokom njihove maturacije koje nisu direktno povezane sa melanogenezom. Pokazano je da se kulture u početnoj fazi (1 dan), konfluentnoj fazi (6 dana) i kasnoj fazi (9 dana) ćelijskog rasta razlikuju na osnovu izoenzimskih spektara leucin-aminopeptidaze i alfa-naftil-acetat-esteraze. Tokom razvitka ćelijskih kultura B-16/C3 zapažene su kvantitativne i kvalitativne promene leucin-aminopeptidaze i alfa-naftil-acetat-esteraze u njihovim izoenzimskim spektrima. Utvrđeno je da postoji porast aktivnosti bakar-cink superoksid-dismutaze tokom starenja kulture. Porast aktivnosti bakar-cink superoksid-dismutaze može biti povezan sa melanogenezom i/ili diferencijacijom B-16/C3 ćelija

A possible protective role of coenzyme Q10 on antioxidant defense enzyme activities in kidneys of rats treated with cadmium

The aim of this study is to establish the effects of cadmium (Cd), coenzyme Q10 (CoQ10) and Cd + CoQ10 on the activities of: Superoxide dismutases (total SOD, manganese containing, Mn SOD, copper zinc containing, CuZn SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione-S-transferase (GST), as well as glutathione reductase (GR) in the kidneys of male Wistar albino rats in comparison with control animals. Significantly increased activities of Tot SOD and CuZn SOD in the kidneys of Cd-treated animals were observed. At the same time, the activity of Mn SOD was significantly decreased in the kidneys of animals treated with Cd, as well as with CoQ10. The activity of CAT was significantly lower in rats exposed to CoQ10 and Cd+CoQ10 in respect to the control animals. GSH-Px and GST activities were significantly decreased in all treated groups of animals (Cd, CoQ10 and Cd+CoQ10), while GR activity was not changed. Presented results reveal, that CoQ10 only partially reversed Cd-induced changes of antioxidant defense enzyme activities in kidney by reversing the effect of Cd on Tot SOD and CuZn SOD activities.Cilj ovog rada bio je da se objasni efekat kadmijuma (Cd), koenzima Q10 (CoQ10) i Cd + CoQ10 na aktivnost: superoksid dismutaza (ukupne, Uk SOD, mangan sadržavajuće, Mn SOD, bakar cink sadržavajuće, CuZn SOD, katalaze (CAT), glutation peroksidaze (GSH-Px), glutation-S-transferaze (GST) kao i glutation reduktaze (GR) u bubrezima mužjaka Wistar albino pacova u poređenju sa kontrolnom grupom životinja. Dobijeni rezultati pokazuju značajno povećanje aktivnosti Uk SOD i CuZn SOD u bubrezima pacova tretiranih sa Cd. U isto vreme aktivnost Mn SOD je bila značajno smanjena u bubrezima pacova tretiranih sa Cd, kao i tretiranih sa Cd+CoQ10. U poređenju sa kontrolnim životinjama, aktivnost CAT je bila značajno smanjena u bubrezima pacova tretiranih sa CoQ10, kao i tretiranih kombinacijom Cd+CoQ10. Aktivnosti GSH-Px i GST su bile značajno smanjene u bubrezima svih tretiranih grupa životinja (Cd, CoQ10 i Cd+CoQ10), dok se aktivnost GR nije statistički značajno menjala. Iz prikazanih rezultata može se zaključiti, da CoQ10 samo delimično utiče na promené aktivnosti antioksidacionih zaštitnih enzima nastale pod uticajem Cd, kao i da vraća aktivnosti Uk SOD i CuZn SOD na normalne vrednosti.nul

A possible protective role of coenzyme Q10 on antioxidant defense enzyme activities in kidneys of rats treated with cadmium

The aim of this study is to establish the effects of cadmium (Cd), coenzyme Q10 (CoQ10) and Cd + CoQ10 on the activities of: Superoxide dismutases (total SOD, manganese containing, Mn SOD, copper zinc containing, CuZn SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione-S-transferase (GST), as well as glutathione reductase (GR) in the kidneys of male Wistar albino rats in comparison with control animals. Significantly increased activities of Tot SOD and CuZn SOD in the kidneys of Cd-treated animals were observed. At the same time, the activity of Mn SOD was significantly decreased in the kidneys of animals treated with Cd, as well as with CoQ10. The activity of CAT was significantly lower in rats exposed to CoQ10 and Cd+CoQ10 in respect to the control animals. GSH-Px and GST activities were significantly decreased in all treated groups of animals (Cd, CoQ10 and Cd+CoQ10), while GR activity was not changed. Presented results reveal, that CoQ10 only partially reversed Cd-induced changes of antioxidant defense enzyme activities in kidney by reversing the effect of Cd on Tot SOD and CuZn SOD activities.Cilj ovog rada bio je da se objasni efekat kadmijuma (Cd), koenzima Q10 (CoQ10) i Cd + CoQ10 na aktivnost: superoksid dismutaza (ukupne, Uk SOD, mangan sadržavajuće, Mn SOD, bakar cink sadržavajuće, CuZn SOD, katalaze (CAT), glutation peroksidaze (GSH-Px), glutation-S-transferaze (GST) kao i glutation reduktaze (GR) u bubrezima mužjaka Wistar albino pacova u poređenju sa kontrolnom grupom životinja. Dobijeni rezultati pokazuju značajno povećanje aktivnosti Uk SOD i CuZn SOD u bubrezima pacova tretiranih sa Cd. U isto vreme aktivnost Mn SOD je bila značajno smanjena u bubrezima pacova tretiranih sa Cd, kao i tretiranih sa Cd+CoQ10. U poređenju sa kontrolnim životinjama, aktivnost CAT je bila značajno smanjena u bubrezima pacova tretiranih sa CoQ10, kao i tretiranih kombinacijom Cd+CoQ10. Aktivnosti GSH-Px i GST su bile značajno smanjene u bubrezima svih tretiranih grupa životinja (Cd, CoQ10 i Cd+CoQ10), dok se aktivnost GR nije statistički značajno menjala. Iz prikazanih rezultata može se zaključiti, da CoQ10 samo delimično utiče na promené aktivnosti antioksidacionih zaštitnih enzima nastale pod uticajem Cd, kao i da vraća aktivnosti Uk SOD i CuZn SOD na normalne vrednosti.nul

The effect of diethyldithicarbarnate on antioxidant enzyme activities in the blood of rats

Diethyldithiocarbamate (DDC) exhibits a variety of effects including neurotoxic, radio-protective and sensitizing activity. It is a potent copper chelating agent used for the treatment of oxygen toxicity, as an imunomodulator in cancer therapy, as well as in HIV infected patients. In this study we examined the effect of DDC, a superoxide dismutase (SOD) inhibitor, on the activities of copper-zinc containing superoxide dismutase (CuZn SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and glutathione-S-transferase (GST). Three hours after diethyldithiocar-bamate treatment (1 g/kg b.m, i.p) a significant decrease of SOD and increase of GR and GST activities were found in the blood of rats. A negative correlation between SOD and GR and a positive correlation between GR and GST activities were also obtained. DDC induced a concentration dependent increase of GR activity and NADPH consumption in an enzymatic assay in vitro. The obtained results may be interpreted to indicate that a decrease in SOD activity in the blood of rats, after DDC administration, may be compensated for by changes in the activity of some other compounds suggesting that regulation of antioxidative defence is very complex.Dietilditiokarbamat (DDC) ispoljava različite uticaje kao što su neurotoksičnost, radioprotekcija i senzibilizacja. On je snažan helirajući agens za bakar i koristi se u tretiranju kiseonične toksičnosti imunomodulaciji kao i kod pacijenata zaraženih HIV virusom. U ovom radu smo proučavali uticaj DDC, inhibitora superoksid dismutaze (SOD), na aktivnost bakar cink sadržavajuće superoksid dismutaze (CuZn SOD), katalaze (CAT) glutation peroksidaze (GSH-Px), glutation reduktaze (GR) i glutation-S-transferaze (GST).Tretiranje pacova dietilditiokarbamatom u trajanju od tri časa (1g/kgt.m, i.p) dovodi do značajnogsmanjenja aktivnosti CuZn SOD i povećanja aktivnosti GR i GST u krvi pacova. Takodje je dobijena negativna korelacija izmedju SOD i GR aktivnosti. DDC indukuje povećanje GR aktivnosti koje je koncentraciono zavisno kao i NADPH potrošnju u in vitro enzimatskom eseju. Dobijeni rezultati ukazuju na činjenicu da smanjivanje aktivnosti SOD u krvi pacova posle tretiranja DDCom može biti kompenzovano promenom aktivnosti drugih komponenti što dalje sugeriše da je regulacija antioksidacione zaštite veoma kompleksna.nul

A possible protective role of coenzyme Q10 on antioxidant defense system in the heart of rats treated with cadmium

The effect of cadmium (Cd), coenzyme Q10 (CoQ10) and Cd+CoQ10 on the activities of superoxide dismutases (total SOD), manganese containing superoxide dismutase (Mn SOD) and copper-zinc containing superoxide dismutase (Cu,Zn SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione-Stransferase (GST), glutathione reductase (GR) and concentrations of ascorbic acid (AsA) and vitamin E (Vit E) in the heart of male Wistar albino rats were studied in comparison to the controls and cadmium treated animals. Cd induces a significant increase of total SOD, Cu, Zn SOD and GSH-Px activities, as well as AsA and Vit E concentrations, but leads to a significant decrease of CAT and GR activities. CoQ10 induces a significant increase of total SOD, Mn SOD, Cu, Zn SOD and GSH-Px activities, as well as AsA and Vit E concentrations. In the same group of animals the activities of CAT, GST and GR were significantly decreased. By concomitant treatment of rats with Cd+CoQ10 the activities of total SOD, Mn SOD and GSH-Px, as well as concentrations of AsA and Vit E were markedly increased. In the same group of animals the activities of Cu, Zn SOD, CAT and GR were significantly decreased. In respect to the Cd treated rats in Cd+CoQ10 partially are reversed changes (Cu,Zn SOD) of antioxidant defense system in the heart.Ispitivan je uticaj kadmijuma (Cd), koenzima Q10 (CoQ10) i Cd+ CoQ10 na aktivnost ukupne superoksid dismutaze (Uk SOD), mangan sadržavajuće superoksid dismutaze (Mn SOD), bakar cink sadržavajuće superoksid dismutaze (Cu,Zn SOD), katalaze (CAT), glutation peroksidaze (GSH-Px), glutation-Stransferaze (GST) i glutation reduktaze (GR), kao i na koncentracije askorbinske kiseline (AsA) i vitamina E (Vit E) u srcu mužjaka Wistar albino pacova u poređenju sa kontrolnom grupom i grupom tretiranom sa kadmijumom. Cd u srcu indukuje značajno povećanje aktivnosti Uk SOD, Cu,Zn SOD i GSH-Px, kao i koncentracija AsA Vit E uz značajno smanjenje aktivnosti CAT i GR. CoQ10 izaziva značajno povećanje aktivnosti Uk SOD, Mn SOD, Cu,Zn SOD i GSH-Px, kao i koncentracija AsA i Vit E. U istoj grupi životinja aktivnosti CAT, GST i GR su značajno smanjene. Istovremeni tretman sa Cd+CoQ10 dovodi do značajnog povećanja aktivnosti Uk SOD, Mn SOD i GSH-Px, kao i koncentracija AsA i Vit E. U istoj grupi životinja aktivnosti Cu,Zn SOD, CAT i GR su značajno smanjene. U poređenju sa pacovima tretiranim sa kadmijumom kod Cd+CoQ10 grupe dobijen je delimični zaštitni efekat (Cu,Zn SOD) na antioksidacioni zaštitni sistem u srcu pacova.nul

Effect of olive oil on antioxidative defense components in liver of cadmium intoxicated rats

The present study investigated the possible protective role of olive oil on antioxidative defense components in liver of cadmium (Cd) intoxicated animals. Male, Wistar albino 3 months old rats were injected with a single dose of: 1) CdCl2 (0.4 mg Cd/kg i.p. and sacrificed after 24h), 2) olive oil (0.4 ml olive oil/kg b.m., i.m., 48h before the sacrificing) and 3) olive oil + Cd (in above mentioned amounts). Treatment with Cd increased lipid peroxidation (LP) in blood and activities of alanine and aspartate transaminase (ALT and AST) in plasma. Concentrations of non enzymatic components of antioxidative defense system (AOS): vitamin C (Vit C) and vitamin E (Vit E) were significantly decreased in liver tissue of Cd treated rats. Pretreatment with olive oil reversed Cd induced alterations in LP content and transaminases activities, as well as on endogenous antioxidive defense components.U ovom radu ispitivana je moguća zaštitna uloga maslinovog ulja na antioksidacione zaštitne komponente u jetri pacova intoksiciranim kadmijumom (Cd). Mužjaci pacova Wistar albino, starosti tri meseca akutno su tretirani sa: 1) CdCl (0.4 mg Cd/kg t.m., i.p., 24h pre žrtvovanja), 2) maslinovim uljem (0.4 ml maslinovog ulja/kg t.m., i.m., 48h pre žrtvovanja) i 3) maslinovim uljem + Cd (u navedenim količinama). Posle tretmana kadmijumom značajno se povećava koncentracija lipidnih peroksida (LP) u krvi, kao i aktivnost alanin i aspartat transaminaza (ALT i AST) u plazmi. Koncentracije neenzimskih komponenti antioksidacionog zaštitnog sistema (AOS): vitamin C (Vit C) i vitamin E (Vit E) su značajno smanjene u jetri kod pacova tretiranih kadmijumom. Eksperimenti sa pacovima koji su u pred tretmanu dobijali maslinovo ulje pokazuju da maslinovo ulje sprečava ili ublažava kadmijumom izazvane promene u koncentraciji LP, aktivnosti, kao i koncentraciji endogenih antioksidacionih zaštitnih komponenti

Effect of olive oil on antioxidative defense components in liver of cadmium intoxicated rats

The present study investigated the possible protective role of olive oil on antioxidative defense components in liver of cadmium (Cd) intoxicated animals. Male, Wistar albino 3 months old rats were injected with a single dose of: 1) CdCl2 (0.4 mg Cd/kg i.p. and sacrificed after 24h), 2) olive oil (0.4 ml olive oil/kg b.m., i.m., 48h before the sacrificing) and 3) olive oil + Cd (in above mentioned amounts). Treatment with Cd increased lipid peroxidation (LP) in blood and activities of alanine and aspartate transaminase (ALT and AST) in plasma. Concentrations of non enzymatic components of antioxidative defense system (AOS): vitamin C (Vit C) and vitamin E (Vit E) were significantly decreased in liver tissue of Cd treated rats. Pretreatment with olive oil reversed Cd induced alterations in LP content and transaminases activities, as well as on endogenous antioxidive defense components.U ovom radu ispitivana je moguća zaštitna uloga maslinovog ulja na antioksidacione zaštitne komponente u jetri pacova intoksiciranim kadmijumom (Cd). Mužjaci pacova Wistar albino, starosti tri meseca akutno su tretirani sa: 1) CdCl (0.4 mg Cd/kg t.m., i.p., 24h pre žrtvovanja), 2) maslinovim uljem (0.4 ml maslinovog ulja/kg t.m., i.m., 48h pre žrtvovanja) i 3) maslinovim uljem + Cd (u navedenim količinama). Posle tretmana kadmijumom značajno se povećava koncentracija lipidnih peroksida (LP) u krvi, kao i aktivnost alanin i aspartat transaminaza (ALT i AST) u plazmi. Koncentracije neenzimskih komponenti antioksidacionog zaštitnog sistema (AOS): vitamin C (Vit C) i vitamin E (Vit E) su značajno smanjene u jetri kod pacova tretiranih kadmijumom. Eksperimenti sa pacovima koji su u pred tretmanu dobijali maslinovo ulje pokazuju da maslinovo ulje sprečava ili ublažava kadmijumom izazvane promene u koncentraciji LP, aktivnosti, kao i koncentraciji endogenih antioksidacionih zaštitnih komponenti

Metabolism of SIN-1: Peroxynitrite donor in rat red blood cells

The aim of this study is elucidation of the molecular mechanism of the 3-morpholino-sydnonimine-hydrochloride (SIN-1) action, particularly its metabolism in rat erythrocytes and reticulocytes. SIN-1 is active metabolite of molsidomine, an established vasodilator drug, clinically used in the treatment of coronary artery disease. Rats erythrocyte and reticulocyte-rich suspensions were aerobically incubated without (control) or in the presence of SIN-1 (0.1, 0.25, 0.5, 1,0 and 1.5 mM). The concentrations of reactive nitrogen species (RNS), reactive oxygen species (ROS) and parameters that indicate oxidative damage of red blood cells were determined after incubation. In rat erythrocytes SIN-1 in high doses increased concentrations of nitrite (NO+ ion indicator), hydroxylamine (NO- ion indicator) and 3-nitrotyrosine (peroxynitrite indicator). Concentration of superoxide anion radical (O2) increased in the presence of SIN-1 on dose-dependent way (except in the presence of higher applied dose), while level of hydrogen peroxide (H2O2) did not alter. In rat reticulocytes SIN-1 significantly increased 3-nitrotyrosine concentrations only. In addition, low doses of SIN-1 induced decrease of O2.- level, while concentration of H2O2 increased dose-dependently. These data indicate that SIN-1-induced nitrosative and oxidative stress in rat red blood cells. The strong oxidative damage of erythrocytes and reticulocytes (increased level of methemoglobin, Heinz bodies and lipid peroxides concentrations) are appeared as consequence of SIN-1-induced oxidative stress.Cilj ovog rada je da se objasne molekularni mehanizmi delovanja 3-morfolino-sidnonimin-hidrohlorida (SIN-1) sa posebnim akcentom na metabolizam ovog leka u eritrocitima i retikulocitima pacova. SIN-1 je aktivni metabolit molsidomina, leka koji se klinički koristi u terapiji bolesti koronarnih arterija. Suspenzije eritrocita i crvenih krvnih ćelija bogate retikulocitima su aerobno inkubirane bez (kontrola) ili u prisustvu različitih koncentracija SIN-1 (0.1, 0.25, 0.5, 1.0 i 1.5 mM). Koncentracije reaktivnih vrsta azota (RNS) i reaktivnih vrsta kiseonika (ROS), kao i koncentracije parametara koji ukazuju na oksidaciona oštećenja crvenih krvnih ćelija, određivane su nakon inkubacije. U eritrocitima pacova, SIN-1 samo u visokim dozama povećava koncentracije nitrita (indikator NO+ jona) hidroksilamina (indicator NO- jona) i 3-nitrotirozina (indikator peroksinitrita). Koncentracija superoksid anjon radikala (O2.-) je povećana na dozno-zavisan način (osim u prisustvu maksimalne primenjene doze SIN-1) dok se nivo vodonik peroksida (H2O2) ne menja. U retikulocitima pacova SIN-1 značajno povećava samo koncentraciju 3-nitrotirozina. Niske doze SIN-1 indukuju smanjenje O2.- nivoa, dok koncentracije H2O2 rastu na dozno-zavisan način. Na osnovu iznetih podataka, možemo zaključiti da SIN-1 indukuje nitrozacioni i oksidacioni stres u crvenim krvnim ćelijama pacova. Ovakvo stanje je posledično praćeno snažnim oksidacionim oštećenjem eritrocita i retikulocita (povećanjem koncentracija methemoglobina, Heinz-ovih telašaca i lipidnih peroksida).nul

Metabolism of SIN-1: Peroxynitrite donor in rat red blood cells

The aim of this study is elucidation of the molecular mechanism of the 3-morpholino-sydnonimine-hydrochloride (SIN-1) action, particularly its metabolism in rat erythrocytes and reticulocytes. SIN-1 is active metabolite of molsidomine, an established vasodilator drug, clinically used in the treatment of coronary artery disease. Rats erythrocyte and reticulocyte-rich suspensions were aerobically incubated without (control) or in the presence of SIN-1 (0.1, 0.25, 0.5, 1,0 and 1.5 mM). The concentrations of reactive nitrogen species (RNS), reactive oxygen species (ROS) and parameters that indicate oxidative damage of red blood cells were determined after incubation. In rat erythrocytes SIN-1 in high doses increased concentrations of nitrite (NO+ ion indicator), hydroxylamine (NO- ion indicator) and 3-nitrotyrosine (peroxynitrite indicator). Concentration of superoxide anion radical (O2) increased in the presence of SIN-1 on dose-dependent way (except in the presence of higher applied dose), while level of hydrogen peroxide (H2O2) did not alter. In rat reticulocytes SIN-1 significantly increased 3-nitrotyrosine concentrations only. In addition, low doses of SIN-1 induced decrease of O2.- level, while concentration of H2O2 increased dose-dependently. These data indicate that SIN-1-induced nitrosative and oxidative stress in rat red blood cells. The strong oxidative damage of erythrocytes and reticulocytes (increased level of methemoglobin, Heinz bodies and lipid peroxides concentrations) are appeared as consequence of SIN-1-induced oxidative stress.Cilj ovog rada je da se objasne molekularni mehanizmi delovanja 3-morfolino-sidnonimin-hidrohlorida (SIN-1) sa posebnim akcentom na metabolizam ovog leka u eritrocitima i retikulocitima pacova. SIN-1 je aktivni metabolit molsidomina, leka koji se klinički koristi u terapiji bolesti koronarnih arterija. Suspenzije eritrocita i crvenih krvnih ćelija bogate retikulocitima su aerobno inkubirane bez (kontrola) ili u prisustvu različitih koncentracija SIN-1 (0.1, 0.25, 0.5, 1.0 i 1.5 mM). Koncentracije reaktivnih vrsta azota (RNS) i reaktivnih vrsta kiseonika (ROS), kao i koncentracije parametara koji ukazuju na oksidaciona oštećenja crvenih krvnih ćelija, određivane su nakon inkubacije. U eritrocitima pacova, SIN-1 samo u visokim dozama povećava koncentracije nitrita (indikator NO+ jona) hidroksilamina (indicator NO- jona) i 3-nitrotirozina (indikator peroksinitrita). Koncentracija superoksid anjon radikala (O2.-) je povećana na dozno-zavisan način (osim u prisustvu maksimalne primenjene doze SIN-1) dok se nivo vodonik peroksida (H2O2) ne menja. U retikulocitima pacova SIN-1 značajno povećava samo koncentraciju 3-nitrotirozina. Niske doze SIN-1 indukuju smanjenje O2.- nivoa, dok koncentracije H2O2 rastu na dozno-zavisan način. Na osnovu iznetih podataka, možemo zaključiti da SIN-1 indukuje nitrozacioni i oksidacioni stres u crvenim krvnim ćelijama pacova. Ovakvo stanje je posledično praćeno snažnim oksidacionim oštećenjem eritrocita i retikulocita (povećanjem koncentracija methemoglobina, Heinz-ovih telašaca i lipidnih peroksida).nul