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Influence of butilated hidroxytoluene, tocopherol and probucol on free radical reactions
ANTIOXIDANT EFFECTS OF CERTAIN D U R I N G E X T R E M A L CONDITIONS
PHARMACOLOGIC
AGENTS
V. Kip.iaDi, T. Sanikidze, N. Antelava, A. Antelava Tbilisi State M e d i c a l University, Asatiani 7, 3 8 0 0 7 7 Tbilisi. Republic of Georgia W i t h the a i m of f i n d i n g out and to b e c o m e c l e a r the m e c h a n i s m s of the action of p h a r m a c o l o g i c a l d e f e n s i v e agents of the body from free radical injury d u r i n g the c e r t a i n e x t r e m a l states, it was studied the i n f l u e n c e of Ionol, V e r a p a m i l and O l i g o c r i n on the p r o c e s s e s of peroxide oxidation during traumatic shock and hemicastration. The paramagnetic centers of myocardium, liver, K i d n e y a n d b l o o d of r a t s was e x a m i n e d w i t h electron spin resonance (EPR). The r e s u l t s of s p e c t r o s c o p i c e x a m i n a t i o n showed, that the p r o c e s s e s of p e r o x i d e o x i d a t i o n in a n i m a l s d u r i n g traumatic shock and hemicastration are s t i m u l a t e d , which has been expressed in disturbances of m i t o c h o n d r i a l e l e c t r o n transport chain on the site of NAD.H: U b i q u i n o n oxidoreductase, a c c u m u l a t i o n of Mn contenting complexes and inactivation of e n z y m e superoxidedismutase (SOD) in the organs, appearance of methemoglobin and i n c r e a s e o f oxidized ceruloplasmin in the b l o o d . Ionol, Verapamil and Oligocrin render unidirection normalization effects on the c h a r a c t e r i s t i c standars of p e r o x i d e o x i d a t i o n and the condition of mitochondrial electron transport chain in organs during the extremal states. We can suggest, that the general core of a n t i o x i d a n t a c t i o n of e x a m i n e d preparatio'ns is a v o i d a n c e of e x c e s s i v e n o x i o u s affect of c y t o z o l i c c a l c i u m due tc membrane - stabilization influences.
CAN HIGH DOSES OF VITAMIN E MODIFY PARAMETERS OF STREPTOZOTOCIN DIABETES IN RATS? J. KopeckS, ~Z. ~Tervinkova, +V. Pali~ka, J. Zoulovfi, M. Miillerovfi, J. Kvfitina, Institute of Exp. Biopharmaceutics, Joint Research Center of PRO.MED.CS Praha a.s. & the Academy of Sciences of the Czech Republic, Heyrovsk6ho 1207, CZ-50002 Hradec Krfilov6; "Faculty of Medicine, Charles University, Hradec Krfilov6;" Department of Clin. Biochemistry, Faculty Hospital, Hradec Krfilov6. Czech Republic. Pretreatment with c~-tocopherol acetate provides complete protection against the diabetogenic effect of alloxan and substantially reduces diabetogenic effect of streptozotocin (STZ) on the free radical scavenger basis. Since there is growing evidence that nonenzymatic glycation of proteins can also lead to the formation of free radicals we decided to investigate the effects of vitamin E in overt diabetes mellitus. Diabetes was induced in 16 male Wistar rats (220-300 g) by i.p. administration of 50 mg/kg STZ (Zanosar ®, Upjohn Co.). 16 control rats received i.p. 0.9 % NaCI. 4 weeks later 8 STZ rats and 8 control rats started to be treated With DL-a-tocopherol acetate (~TCA), and 8 STZ rats and 8 control rats with 0.9 % NaCI. Drugs were administered orally three times per week for 4 weeks. The single doses of ct-TCA were 400 mg/kg. Afterwards rats were sacrified, blood analysed and organs weighed. Presented results are mean ±SD. STZ induced statist, significant elevation in plasma levels of glucose (20.1_+7.9 in STZ rats vs 8.8+_l.6 mmol/l in controls), glycohaemoglobin (3.4_-/O.5 % vs 1.0_+0.6 %), urea (10.0+~.6 vs 6.8_+0.8 mmol/l), tot. cholesterol (2.2_+0.4 vs 1.4_+0.1 mmol/I) and fall in creatininaemia (37+A3 vs 43-+1 l~mol/I). Body weight was decreased and kidney weight increased in STZ rats. Administratin of ~-TCA had no effect on investigated parameters both in control and STZ rats, except of partial restoring elavated plasma cholesterol levels in STZ rats (1.8 _+0.2 mmol/l). With respect to progressive character of diabetic complications we suppose that long-term study would be useful.
- - 220
CUMOL HYDROPEROXIDE INFLUENCE ON GLUTATHIONE METABOLISM L.S. Kolesnichenko, Yu.V. Ryzhenkova, E.F. Glushkova Irkutsk State Medical University, U1. Krasnogo Vosstanija, 1, 664003, Irkutsk, Russia. Majority of glutathione metabolizing enzymes (GME) are inducible, but possibility of glutathione peroxidase (GPx) induction is questionable. The single injection of cumol hydroperoxide (CuOOH) 66-329 l*Mol/kg almost does not influence on activities of GME; only glutathione S-transferase (GST) increases in kidney cytosol and glutathione reductase (GR) decreases i n liver mitochondria. 10 everyday injections increase GSH concentration in liver and kidney cytosol, but decrease one in liver mitochondria; GPx in liver cytosol and GST in liver mitoehondria increase, GST in heart cytosol decreases (all these results are within 20-50%). Increase of GPx and GSH in liver cytosol suggests possibility of GPx and enzymes of GSH biosynthesis induction at peroxidative stress, what is optimal for defense from peroxidation. Phenobarbital and butylated hydroxytoluene induce GST and GR, but not GPx. Changes of GSH metabolism are different in cytosol of investigated organs and in cytosol and mitochondria of liver. It suggests, that CuOOH course protects liver cytosol better, than mitochondria, and liver better than other organs. No induction is in the heart.
INFLUENCE OF BUTILATED H I D R O X Y T O L U E N E , TOCOPHEROL AND P R O B U C O L ON FREE RADICAL REACTIONS. V.A. Kosolapov. O.V. Ostrovsky The Volgograd Medical Academy, Dep. Pharmacology, Pl. Pavshikh Bortsov. 1, 400066 Volgograd, Russia. The effects of lipid-soluble drugs butilated hidroxytoluene (BHT), probucol and ~-tocopherol were investigated on three models with free radical reactions. The drug concentrations, which inhibited reaction by 50% (IC50) were determined. Tocopherol and BHT IC50 in reaction of askorbat-indueed lipid peroxidation in rat's liver microsomes were 0.5 and 0.8 ~M/1, while probucol IC50 was 5.6 g M / 1 only. Determination of effects of the drugs in the reaction with super-oxide anion generation (reaction of quercetin oxidation) showed that tocopherol and probucol [C50 were 13 and 18 ~xM/1, while BHT IC50 was much higher and made 500 IxM/I. Tocopherol IC50 for the reaction with free radical diphenil-picryl-hidrazile was 14 ~xM/I. As for the other drugs, BHT and probncol IC50 made 500 and 100 ~tM/[. These results indicated that investigated drugs displayed different effects on the various models with free radical reactions. The antihypoxm and antiishaemic effects of the drugs are investigated.
PHARMACOLOGIC
AGENTS
V. Kip.iaDi, T. Sanikidze, N. Antelava, A. Antelava Tbilisi State M e d i c a l University, Asatiani 7, 3 8 0 0 7 7 Tbilisi. Republic of Georgia W i t h the a i m of f i n d i n g out and to b e c o m e c l e a r the m e c h a n i s m s of the action of p h a r m a c o l o g i c a l d e f e n s i v e agents of the body from free radical injury d u r i n g the c e r t a i n e x t r e m a l states, it was studied the i n f l u e n c e of Ionol, V e r a p a m i l and O l i g o c r i n on the p r o c e s s e s of peroxide oxidation during traumatic shock and hemicastration. The paramagnetic centers of myocardium, liver, K i d n e y a n d b l o o d of r a t s was e x a m i n e d w i t h electron spin resonance (EPR). The r e s u l t s of s p e c t r o s c o p i c e x a m i n a t i o n showed, that the p r o c e s s e s of p e r o x i d e o x i d a t i o n in a n i m a l s d u r i n g traumatic shock and hemicastration are s t i m u l a t e d , which has been expressed in disturbances of m i t o c h o n d r i a l e l e c t r o n transport chain on the site of NAD.H: U b i q u i n o n oxidoreductase, a c c u m u l a t i o n of Mn contenting complexes and inactivation of e n z y m e superoxidedismutase (SOD) in the organs, appearance of methemoglobin and i n c r e a s e o f oxidized ceruloplasmin in the b l o o d . Ionol, Verapamil and Oligocrin render unidirection normalization effects on the c h a r a c t e r i s t i c standars of p e r o x i d e o x i d a t i o n and the condition of mitochondrial electron transport chain in organs during the extremal states. We can suggest, that the general core of a n t i o x i d a n t a c t i o n of e x a m i n e d preparatio'ns is a v o i d a n c e of e x c e s s i v e n o x i o u s affect of c y t o z o l i c c a l c i u m due tc membrane - stabilization influences.
CAN HIGH DOSES OF VITAMIN E MODIFY PARAMETERS OF STREPTOZOTOCIN DIABETES IN RATS? J. KopeckS, ~Z. ~Tervinkova, +V. Pali~ka, J. Zoulovfi, M. Miillerovfi, J. Kvfitina, Institute of Exp. Biopharmaceutics, Joint Research Center of PRO.MED.CS Praha a.s. & the Academy of Sciences of the Czech Republic, Heyrovsk6ho 1207, CZ-50002 Hradec Krfilov6; "Faculty of Medicine, Charles University, Hradec Krfilov6;" Department of Clin. Biochemistry, Faculty Hospital, Hradec Krfilov6. Czech Republic. Pretreatment with c~-tocopherol acetate provides complete protection against the diabetogenic effect of alloxan and substantially reduces diabetogenic effect of streptozotocin (STZ) on the free radical scavenger basis. Since there is growing evidence that nonenzymatic glycation of proteins can also lead to the formation of free radicals we decided to investigate the effects of vitamin E in overt diabetes mellitus. Diabetes was induced in 16 male Wistar rats (220-300 g) by i.p. administration of 50 mg/kg STZ (Zanosar ®, Upjohn Co.). 16 control rats received i.p. 0.9 % NaCI. 4 weeks later 8 STZ rats and 8 control rats started to be treated With DL-a-tocopherol acetate (~TCA), and 8 STZ rats and 8 control rats with 0.9 % NaCI. Drugs were administered orally three times per week for 4 weeks. The single doses of ct-TCA were 400 mg/kg. Afterwards rats were sacrified, blood analysed and organs weighed. Presented results are mean ±SD. STZ induced statist, significant elevation in plasma levels of glucose (20.1_+7.9 in STZ rats vs 8.8+_l.6 mmol/l in controls), glycohaemoglobin (3.4_-/O.5 % vs 1.0_+0.6 %), urea (10.0+~.6 vs 6.8_+0.8 mmol/l), tot. cholesterol (2.2_+0.4 vs 1.4_+0.1 mmol/I) and fall in creatininaemia (37+A3 vs 43-+1 l~mol/I). Body weight was decreased and kidney weight increased in STZ rats. Administratin of ~-TCA had no effect on investigated parameters both in control and STZ rats, except of partial restoring elavated plasma cholesterol levels in STZ rats (1.8 _+0.2 mmol/l). With respect to progressive character of diabetic complications we suppose that long-term study would be useful.
- - 220
CUMOL HYDROPEROXIDE INFLUENCE ON GLUTATHIONE METABOLISM L.S. Kolesnichenko, Yu.V. Ryzhenkova, E.F. Glushkova Irkutsk State Medical University, U1. Krasnogo Vosstanija, 1, 664003, Irkutsk, Russia. Majority of glutathione metabolizing enzymes (GME) are inducible, but possibility of glutathione peroxidase (GPx) induction is questionable. The single injection of cumol hydroperoxide (CuOOH) 66-329 l*Mol/kg almost does not influence on activities of GME; only glutathione S-transferase (GST) increases in kidney cytosol and glutathione reductase (GR) decreases i n liver mitochondria. 10 everyday injections increase GSH concentration in liver and kidney cytosol, but decrease one in liver mitochondria; GPx in liver cytosol and GST in liver mitoehondria increase, GST in heart cytosol decreases (all these results are within 20-50%). Increase of GPx and GSH in liver cytosol suggests possibility of GPx and enzymes of GSH biosynthesis induction at peroxidative stress, what is optimal for defense from peroxidation. Phenobarbital and butylated hydroxytoluene induce GST and GR, but not GPx. Changes of GSH metabolism are different in cytosol of investigated organs and in cytosol and mitochondria of liver. It suggests, that CuOOH course protects liver cytosol better, than mitochondria, and liver better than other organs. No induction is in the heart.
INFLUENCE OF BUTILATED H I D R O X Y T O L U E N E , TOCOPHEROL AND P R O B U C O L ON FREE RADICAL REACTIONS. V.A. Kosolapov. O.V. Ostrovsky The Volgograd Medical Academy, Dep. Pharmacology, Pl. Pavshikh Bortsov. 1, 400066 Volgograd, Russia. The effects of lipid-soluble drugs butilated hidroxytoluene (BHT), probucol and ~-tocopherol were investigated on three models with free radical reactions. The drug concentrations, which inhibited reaction by 50% (IC50) were determined. Tocopherol and BHT IC50 in reaction of askorbat-indueed lipid peroxidation in rat's liver microsomes were 0.5 and 0.8 ~M/1, while probucol IC50 was 5.6 g M / 1 only. Determination of effects of the drugs in the reaction with super-oxide anion generation (reaction of quercetin oxidation) showed that tocopherol and probucol [C50 were 13 and 18 ~xM/1, while BHT IC50 was much higher and made 500 IxM/I. Tocopherol IC50 for the reaction with free radical diphenil-picryl-hidrazile was 14 ~xM/I. As for the other drugs, BHT and probncol IC50 made 500 and 100 ~tM/[. These results indicated that investigated drugs displayed different effects on the various models with free radical reactions. The antihypoxm and antiishaemic effects of the drugs are investigated.