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Drug and Chemical Toxicology Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/idct20
Increased Reduced Glutathione and Glutathione STransferase Activity in Chronic Cephaloridine Nephrotoxicity Studies in the Rat a
a
a
M. F. Olivier , H. Dutertre-Catella1 , M. Thevenin , a
a
C. Martin , J.-M. Warnet & J.-R. Claude
a
a
Laboratoire, de Toxicologie, Faculté de Pharmacie, 4, avenue de I'Observatoire, F 75006, Paris Published online: 11 Apr 2015.
To cite this article: M. F. Olivier, H. Dutertre-Catella1, M. Thevenin, C. Martin, J.M. Warnet & J.-R. Claude (1990) Increased Reduced Glutathione and Glutathione STransferase Activity in Chronic Cephaloridine Nephrotoxicity Studies in the Rat, Drug and Chemical Toxicology, 13:2-3, 209-219 To link to this article: http://dx.doi.org/10.3109/01480549009018121
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or
indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [RMIT University] at 00:04 15 August 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/terms-and-conditions
DRUG AND CHEMICAL TOXICOLOGY, 1 3 ( 2 & 3 ) , 2 0 9 - 2 1 9 (1990)
INCREASED REDUCED GLUTATHIONE AND GLUTATHIONE S-TRANSFERASE ACTIVITY IN CHRONIC CEPHALORIDINE
Downloaded by [RMIT University] at 00:04 15 August 2015
NEPHROTOXlClTY STUDIES IN THE RAT
M.F.Olivier, H.Dutertre-Catella', M.Thevenin, C.Martin, J.-M.Warnet & J.-R.Claude Laboratoire, de Toxicologie, Faculte de ~Pharmacie, 4, avenue de I'Obsewatoire, F 75006 Paris
ABSTRACT
The effect of repeated cephaloridine treatment on renal glutathione and related enzymes has been investigated in young adult male and female Sprague-Dawley rats. Animals were given intraperitoneally daily doses of either 750mg/kg for two weeks or 500 or 750mg/kg for three months. Measurement of blood and urinary parameters (electrolytes, urea, creatinine) did not reveal any renal function impairment and histological examination confirmed the absence of. renal damage. By contrast, an increase in reduced glutathione (2 to 3-fold) and glutathione S-transferase activity (1.5 to 2-fold) was observed. These results are consistent with the development of an adaptative phenomenon to cephaloridine subchronic treatment in the rat, leading to a tolerance to high repeated doses.
110 whom correspondence should be addressed
209
Copyright
0 1990 by Marcel Dekker, Inc.
210
O L I V I E R ET A L .
JNTRODUCTION
The rat is known to cephaloridine
be a relatively resistant species to
(CPH)-induced nephrotoxicity. GOLDSTEIN et al.1
reported that younger rats were less susceptible to very high single doses of CPH than older rats. Also, young rats have been found tolerant to repeated administration of this cephalosporin 2 1 3 ~ 4In .
Downloaded by [RMIT University] at 00:04 15 August 2015
order to explore the biochemical mechanism of this resistance, male and female Sprague-Dawley rats were administered either 14-day or 3-month course of CPH and the renal glutathione content and relatedenzyme activities were examined on account of their critical role in detoxication.
MATERIAL AND MFTHODS
Animals and ExDerimentaI Procedu re ; Groups of six male and six female Sprague-Dawley rats were used. Animals were housed in a ventilated room (temperature: 2225°C; humidity: 40-60%) with a 12-hr lighvdark cycle. They were fed on a commercial diet and water ad libitum. Rats received CPH intraperitoneally at doses of 750 mg/kg per day in the morning, for two weeks, and 500 or 750 mg/kg per day for three months. Controls received saline solution. All the rats were 16 weeks old at termination of the study. Following the last injection, rats were placed in individual metabolism cages for collection of 24 hr-urine samples. They were anesthetized and thoracotomized: the
sus-hepatic vein and the right auricle of the heart were excised for blood collection and liver and kidneys were perfused simultaneously via the
left ventricle with
heparinized saline solution at 37°C.
Exsanguined kidneys were removed immediately and weighed. A small
211
EFFECT OF CEPHALORIDINE TREATMENT ON GLUTATHIONE
part of one kidney was immediately placed in Bouin's solution for ulterior histologic examination; the rest was homogenized at +4"C, in 5 vol. of 5% 5-sulfosalicylic acid. The homogenate was centrifuged at 3000 r.p.rn. for 15 min and stored at -4OOC. Concentrations of total,
reduced and oxidized glutathione were measured in this homogenate. The other kidney was homogenized in 9 volumes of buffered solution pH 7.0 (glucose 5 mM, NaCl 140 mM, KH2P04 10 mM, EDTA Naz 1 mM,
Downloaded by [RMIT University] at 00:04 15 August 2015
N a N 3 1 mM), then centrifuged at 3000 r.p.m. for 15 rnin. The supernatant fraction was collected and centrifuged twice at 15,000 g, at +4"C, for 15 min. Cytosol was collected, then frozen in liquid nitrogen and stored at -80°C until glutathione-related enzyme activities were assayed. Enzvmuria Determination: 5 ml urine filtered on PD 10 columns (Pharmacia) were used 5 . NAG (N-acetyl-P-D-glucosaminidase) and y-GT (gamma glutamyl transferase) activities were determined with reagent kits obtained from Boehringer. AAP
(alanine
aminopeptidase)
activity
was
determined
according to DIENER et a1.6. Biochemical Determinations on Kidnev Tissue: Total, reduced and oxidized glutathione were determined using the enzymatic method of TIETZE 7 as modified by GRlFFlTH 8 . Glutathione-reductase (GSSG-reductase) activity
was
measured
according to CARLBERG & MANNERVIKg. Glutathione transferase (GSH S-transferase) activity was assayed according to WARHLOM et a1.l using
1-chloro-2,4-dinitrobenzene
peroxidase
as
a
substrate.
Glutathione
(GSH-peroxidase) activity was assayed according to
W E N D E L et a1.l 1 using curnene hydroperoxide as a substrate. Glutathione-related
enzyme activities were expressed relative to
protein concentration determined by a modified Lowry method1 2 .
O L I V I E R ET AL.
212
HIStOlOgy;
After fixation,
kidney samples were embedded in paraffin.
Sections of kidney were then stained with Masson's trichrome stain or Halmi's stain for the brush border (PAS, orange G, light green, hematoxylin). St;iistical
analySis;
All the results were expressed as mean
2
standard deviation.
Downloaded by [RMIT University] at 00:04 15 August 2015
The differences between control and treated groups were evaluated by Student's t test and a p value of 0.05 or less was considered significant.
RESULTS Blood and urine parameters (electrolytes, urea, creatinine), urinary activities of brush border enzymes (AAP, y-GT) and lysosomal enzymes (NAG) were not modified in treated rats (data not detailed). Histological study confirmed the absence of renal damage since organization and cytological structure of nephrons were normal in the different areas of the kidney (cortex and medulla) in both treated and control rats. The 14-day and 3-month CPH treatments induced a significant increase in total glutathione concentrations, in males as in females (Table 1). This was due to the increase in reduced glutathione in treated groups. In contrast, oxidized glutathione was not modified significantly, except in the females treated with 750 mglkg for 14 days. Glutathione S-transferase activity was significantly increased in
all
treated
groups.
Glutathione
reductase
activity
was
significantly but slightly increased in the 14-day 750 mglkg CPH treated female rats and in the 3-month 750 mglkg treated male rats. Glutathione peroxidase activity remained unchanged in all treated groups (Table 2).
213
EFFECT O F CEPHALORIDINE TREATMENT ON GLUTATHIONE
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OL IV IER ET A L .
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E F F E C T O F C E P H A L O R I D I N E TREATMENT ON GLUTATHIONE
215
DISCUSSION
Repeated administration of high doses of cephaloridine (CPH) for 14 days (750 mg/kg) or 3 months (500 or 750 mg/kg) did not
induce nephrotoxicity in the rat, as there was no variation in biological indices (blood and urine urea, creatinine and electrolytes and enzymuria), nor in histological appearance. This is consistent
Downloaded by [RMIT University] at 00:04 15 August 2015
with literature data which reveal that the rat tolerates repeated doses of CPH: administration of 250 mglkg per day or 1100 mg/kg per day in divided doses given twice daily for 4 weeks induced moderate proximal tubular injury in few rats while the dose of 1100 mg/kg per day given as a single dose produced more renal damages.
As shown by ATKINSON et a1.2, doses that are nephrotoxic given singly had surprisingly little effect when given daily for 5 days. Later, with
repeated
administration
of nephrotoxic constant or
increasing doses of CPH, WACHSMUTH4 concluded from enzymuria and histology data that an initial phase of cell degeneration and tubular necrosis was followed by a phase of regeneration with young cells which tolerated larger doses of the nephrotoxic compound than the original epithelium. It should be noticed that these experiments were conducted in young rats. Age-related differences in susceptibility to
CPH nephrotoxicity also were evaluated by GOLDSTEIN et al.1 with single doses. This author observed that 2.5 or 4 month-old rats were less susceptible to CPH nephrotoxicity than 10-12 or 27-29 monthold rats. This resistance could be due to weak uptake of CPH in Proximal tubular cells as observed by TUNE13 with single CPH dosage. Nevertheless, in our experiments, in spite of the absence of kidney injury, there was
a significant
alteration
of glutathione
content and glutathione-related enzyme system: repeated 14-day
216
O L I V I E R ET AL.
(750 rnglkg) or 3-month treatments (500 or 750 mglkg) induced an increase in kidney reduced glutathione content, associated with a slight increase in glutathione reductase and a two-fold increase in glutathione S-transferase activities. The lack of increase in oxidized glutathione
is
well
correlated
with
this
slight
increase
in
glutathione reductase activity. It should be emphasized that the rise in glutathione
S-transferase activity
observed after subchronic
Downloaded by [RMIT University] at 00:04 15 August 2015
treatments also occurs precociously as noticed by COJOCEL et a1.l in the renal cortex of rats treated with 1200 mg/kg per day CPH for 2 days. Interestingly, the variation in glutathione content is opposite
to the one observed by KU015~16117with massive single CPH dosage. This author attributed the glutathione depletion to the nephrotoxic response to the probable lipid peroxidation mechanism, confirmed by
COJOCEL et .I."? Glutathione as well _as glutathione S-transferase are assumed to play a critical role in detoxication mechanism in response to a lipoperoxidative processl8. However, such a mechanism did not seem to take place in our experimental conditions, and this was confirmed
by the lack of variation in glutathione peroxidase activity. With repeated CPH dosage, the increased glutathione content, the synthesis of which is known to increase rapidly after an acute depletion' 9, associated with the two-fold increase in glutathione Stransferase, corresponded to a marked increase in detoxication and strongly suggest an adaptative phenomenon to the cephaloridine nephrotoxicity. Furthermore, this is well correlated with the absence of histological and enzymic renal disturbances.
/iCKNOWLEffiF MENTS
We wish to thank Prof L. Olivier for histological interpretation, Roussel Uclaf Laboratories for a grant to Miss M.F. Olivier and
21 7
EFFECT OF CEPHALORIDINE TREATMENT O N GLUTATHIONE
A.D.R.T.E. (Association pour le Developpement de la Recherche en Toxicologie Experimentale) for financial support.
REFERENCFS
1. R.S. Goldstein, D.A. Pasino & J.B. Hook, Cephaloridine nephrotoxicity in aging male Fischer-344 rats, Toxicology,
a,43
Downloaded by [RMIT University] at 00:04 15 August 2015
(1 986). 2. R.M. Atkinson, J.D. Caisey, J.P. Currie, T.R. Middleton, D.A.H. Pratt, H.M. Sharpe & E.G. Tomich, Subacute toxicity of cephaloridine to various species, Toxicol. Appl. Pharmacol., 8, 407 (1966).
3.
F. Silverblatt, W.O. Harrison
& M. Turck, Nephrotoxicity of
cephalosporin antibiotics in experimental animals, J. Infect. Dis.,
128,S 4.
367 (1973).
E.D.Wachsmuth, Adaptation to nephrotoxic effects of cephaloridine in subacute rat toxicity studies, Toxicol. Appl. Pharmacol.,
5.
U, 446
(1982).
A. Berscheid, H. Grotsh, M. Hropot & E. Klauss, Enzymuria of the rat: the preparation of urine for enzyme analysis, J. Clin. Chem. Clin. Biochem.,
6.
U. Diener,
a,799 (1983).
E. Knoll, B. Langer, H. Rautenstrauch, D. Ratge & H.
Wisser, Urinary excretion of N-acetyl-P-D-glucosaminidase and alanine aminopeptidase in patients receiving amikacine or cisplatinum, Clin. Chim. Acta,
7.
112,149
(1981).
F. Tietze, Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione. Applications
to mammalian blood and other tissues, Anal. Biochem., 22, 502 (1 969).
OLIVIER ET A L .
218
8.
O.W. Griffith, Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine, Anal. Biochem.,
9.
m, 207
(1980).
I. Carlberg & B. Mannervik, Glutathione reductase, Methods Enzymol.,
U, 484
(1985).
10. M. Warholm, C. Guthenberg, C. Von Bahr & B. Mannervik, Glutathione transferases from human liver, Methods Enzymol.,
Downloaded by [RMIT University] at 00:04 15 August 2015
m, 499
(1985).
11. A. Wendel, Glutathione peroxidase, Methods Enzymol.,
U ,325
(1981). 12. M.A. Markwell, S.M. Hass, L.L. Bieber & N.E. Tolbert, A
modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples, Anal. Biochem., 13.
Bz, 206
(1978).
B.M. Tune, Relationships between the transport and toxicity of cephalosporins in the kidney, J. Infect. Dis.,
m,189
(1975).
14. C. Cojocel, W. Kramer & D. Mayer, Depletion of cytochrome P-450
and alterations in activities of drug metabolizing enzymes induced by cephaloridine in the rat kidney cortex, Biochem. Pharmacol.,
Z, 3781
(1988).
15. C.H. Kuo & J.B. Hook, Depletion of renal cortical glutathione and nephrotoxicity by cephaloridine, cephalothin and gentamicin in male Sprague-Dawley rats, Life Sci., Z, 255 (1982). 16. C.H. Kuo & J.B. Hook, Depletion of renal glutathione content and nephrotoxicity of cephaloridine in rabbits, rats and mice, Toxicol. Appl. Pharmacol.,
B,292
(1982).
17. C.H. Kuo, K. Maita, S.D. Sleight & J.B. Hook, Lipid peroxidation: a possible mechanism of cephaloridine-induced nephrotoxicity, Toxicol. Appl. Pharmacol.,
U , 78 (1983).
EFFECT O F CEPHALORIDINE TREATMENT O N GLUTATHIONE
18.
N. Kaplowitz, Physiological significance of glutathione Stransferases, Am. J. Physiol.,
23-9,G
439 (1980).
19. A. Meister & M.E. Anderson, Glutathione, Ann. Rev. Biochern., 711 (1983).
Downloaded by [RMIT University] at 00:04 15 August 2015
219
2,
Drug and Chemical Toxicology Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/idct20
Increased Reduced Glutathione and Glutathione STransferase Activity in Chronic Cephaloridine Nephrotoxicity Studies in the Rat a
a
a
M. F. Olivier , H. Dutertre-Catella1 , M. Thevenin , a
a
C. Martin , J.-M. Warnet & J.-R. Claude
a
a
Laboratoire, de Toxicologie, Faculté de Pharmacie, 4, avenue de I'Observatoire, F 75006, Paris Published online: 11 Apr 2015.
To cite this article: M. F. Olivier, H. Dutertre-Catella1, M. Thevenin, C. Martin, J.M. Warnet & J.-R. Claude (1990) Increased Reduced Glutathione and Glutathione STransferase Activity in Chronic Cephaloridine Nephrotoxicity Studies in the Rat, Drug and Chemical Toxicology, 13:2-3, 209-219 To link to this article: http://dx.doi.org/10.3109/01480549009018121
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or
indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [RMIT University] at 00:04 15 August 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/terms-and-conditions
DRUG AND CHEMICAL TOXICOLOGY, 1 3 ( 2 & 3 ) , 2 0 9 - 2 1 9 (1990)
INCREASED REDUCED GLUTATHIONE AND GLUTATHIONE S-TRANSFERASE ACTIVITY IN CHRONIC CEPHALORIDINE
Downloaded by [RMIT University] at 00:04 15 August 2015
NEPHROTOXlClTY STUDIES IN THE RAT
M.F.Olivier, H.Dutertre-Catella', M.Thevenin, C.Martin, J.-M.Warnet & J.-R.Claude Laboratoire, de Toxicologie, Faculte de ~Pharmacie, 4, avenue de I'Obsewatoire, F 75006 Paris
ABSTRACT
The effect of repeated cephaloridine treatment on renal glutathione and related enzymes has been investigated in young adult male and female Sprague-Dawley rats. Animals were given intraperitoneally daily doses of either 750mg/kg for two weeks or 500 or 750mg/kg for three months. Measurement of blood and urinary parameters (electrolytes, urea, creatinine) did not reveal any renal function impairment and histological examination confirmed the absence of. renal damage. By contrast, an increase in reduced glutathione (2 to 3-fold) and glutathione S-transferase activity (1.5 to 2-fold) was observed. These results are consistent with the development of an adaptative phenomenon to cephaloridine subchronic treatment in the rat, leading to a tolerance to high repeated doses.
110 whom correspondence should be addressed
209
Copyright
0 1990 by Marcel Dekker, Inc.
210
O L I V I E R ET A L .
JNTRODUCTION
The rat is known to cephaloridine
be a relatively resistant species to
(CPH)-induced nephrotoxicity. GOLDSTEIN et al.1
reported that younger rats were less susceptible to very high single doses of CPH than older rats. Also, young rats have been found tolerant to repeated administration of this cephalosporin 2 1 3 ~ 4In .
Downloaded by [RMIT University] at 00:04 15 August 2015
order to explore the biochemical mechanism of this resistance, male and female Sprague-Dawley rats were administered either 14-day or 3-month course of CPH and the renal glutathione content and relatedenzyme activities were examined on account of their critical role in detoxication.
MATERIAL AND MFTHODS
Animals and ExDerimentaI Procedu re ; Groups of six male and six female Sprague-Dawley rats were used. Animals were housed in a ventilated room (temperature: 2225°C; humidity: 40-60%) with a 12-hr lighvdark cycle. They were fed on a commercial diet and water ad libitum. Rats received CPH intraperitoneally at doses of 750 mg/kg per day in the morning, for two weeks, and 500 or 750 mg/kg per day for three months. Controls received saline solution. All the rats were 16 weeks old at termination of the study. Following the last injection, rats were placed in individual metabolism cages for collection of 24 hr-urine samples. They were anesthetized and thoracotomized: the
sus-hepatic vein and the right auricle of the heart were excised for blood collection and liver and kidneys were perfused simultaneously via the
left ventricle with
heparinized saline solution at 37°C.
Exsanguined kidneys were removed immediately and weighed. A small
211
EFFECT OF CEPHALORIDINE TREATMENT ON GLUTATHIONE
part of one kidney was immediately placed in Bouin's solution for ulterior histologic examination; the rest was homogenized at +4"C, in 5 vol. of 5% 5-sulfosalicylic acid. The homogenate was centrifuged at 3000 r.p.rn. for 15 min and stored at -4OOC. Concentrations of total,
reduced and oxidized glutathione were measured in this homogenate. The other kidney was homogenized in 9 volumes of buffered solution pH 7.0 (glucose 5 mM, NaCl 140 mM, KH2P04 10 mM, EDTA Naz 1 mM,
Downloaded by [RMIT University] at 00:04 15 August 2015
N a N 3 1 mM), then centrifuged at 3000 r.p.m. for 15 rnin. The supernatant fraction was collected and centrifuged twice at 15,000 g, at +4"C, for 15 min. Cytosol was collected, then frozen in liquid nitrogen and stored at -80°C until glutathione-related enzyme activities were assayed. Enzvmuria Determination: 5 ml urine filtered on PD 10 columns (Pharmacia) were used 5 . NAG (N-acetyl-P-D-glucosaminidase) and y-GT (gamma glutamyl transferase) activities were determined with reagent kits obtained from Boehringer. AAP
(alanine
aminopeptidase)
activity
was
determined
according to DIENER et a1.6. Biochemical Determinations on Kidnev Tissue: Total, reduced and oxidized glutathione were determined using the enzymatic method of TIETZE 7 as modified by GRlFFlTH 8 . Glutathione-reductase (GSSG-reductase) activity
was
measured
according to CARLBERG & MANNERVIKg. Glutathione transferase (GSH S-transferase) activity was assayed according to WARHLOM et a1.l using
1-chloro-2,4-dinitrobenzene
peroxidase
as
a
substrate.
Glutathione
(GSH-peroxidase) activity was assayed according to
W E N D E L et a1.l 1 using curnene hydroperoxide as a substrate. Glutathione-related
enzyme activities were expressed relative to
protein concentration determined by a modified Lowry method1 2 .
O L I V I E R ET AL.
212
HIStOlOgy;
After fixation,
kidney samples were embedded in paraffin.
Sections of kidney were then stained with Masson's trichrome stain or Halmi's stain for the brush border (PAS, orange G, light green, hematoxylin). St;iistical
analySis;
All the results were expressed as mean
2
standard deviation.
Downloaded by [RMIT University] at 00:04 15 August 2015
The differences between control and treated groups were evaluated by Student's t test and a p value of 0.05 or less was considered significant.
RESULTS Blood and urine parameters (electrolytes, urea, creatinine), urinary activities of brush border enzymes (AAP, y-GT) and lysosomal enzymes (NAG) were not modified in treated rats (data not detailed). Histological study confirmed the absence of renal damage since organization and cytological structure of nephrons were normal in the different areas of the kidney (cortex and medulla) in both treated and control rats. The 14-day and 3-month CPH treatments induced a significant increase in total glutathione concentrations, in males as in females (Table 1). This was due to the increase in reduced glutathione in treated groups. In contrast, oxidized glutathione was not modified significantly, except in the females treated with 750 mglkg for 14 days. Glutathione S-transferase activity was significantly increased in
all
treated
groups.
Glutathione
reductase
activity
was
significantly but slightly increased in the 14-day 750 mglkg CPH treated female rats and in the 3-month 750 mglkg treated male rats. Glutathione peroxidase activity remained unchanged in all treated groups (Table 2).
213
EFFECT O F CEPHALORIDINE TREATMENT ON GLUTATHIONE
b
00 0 0 c
%
c pr,
0
N 0
0
0
0
0
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Downloaded by [RMIT University] at 00:04 15 August 2015
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E F F E C T O F C E P H A L O R I D I N E TREATMENT ON GLUTATHIONE
215
DISCUSSION
Repeated administration of high doses of cephaloridine (CPH) for 14 days (750 mg/kg) or 3 months (500 or 750 mg/kg) did not
induce nephrotoxicity in the rat, as there was no variation in biological indices (blood and urine urea, creatinine and electrolytes and enzymuria), nor in histological appearance. This is consistent
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with literature data which reveal that the rat tolerates repeated doses of CPH: administration of 250 mglkg per day or 1100 mg/kg per day in divided doses given twice daily for 4 weeks induced moderate proximal tubular injury in few rats while the dose of 1100 mg/kg per day given as a single dose produced more renal damages.
As shown by ATKINSON et a1.2, doses that are nephrotoxic given singly had surprisingly little effect when given daily for 5 days. Later, with
repeated
administration
of nephrotoxic constant or
increasing doses of CPH, WACHSMUTH4 concluded from enzymuria and histology data that an initial phase of cell degeneration and tubular necrosis was followed by a phase of regeneration with young cells which tolerated larger doses of the nephrotoxic compound than the original epithelium. It should be noticed that these experiments were conducted in young rats. Age-related differences in susceptibility to
CPH nephrotoxicity also were evaluated by GOLDSTEIN et al.1 with single doses. This author observed that 2.5 or 4 month-old rats were less susceptible to CPH nephrotoxicity than 10-12 or 27-29 monthold rats. This resistance could be due to weak uptake of CPH in Proximal tubular cells as observed by TUNE13 with single CPH dosage. Nevertheless, in our experiments, in spite of the absence of kidney injury, there was
a significant
alteration
of glutathione
content and glutathione-related enzyme system: repeated 14-day
216
O L I V I E R ET AL.
(750 rnglkg) or 3-month treatments (500 or 750 mglkg) induced an increase in kidney reduced glutathione content, associated with a slight increase in glutathione reductase and a two-fold increase in glutathione S-transferase activities. The lack of increase in oxidized glutathione
is
well
correlated
with
this
slight
increase
in
glutathione reductase activity. It should be emphasized that the rise in glutathione
S-transferase activity
observed after subchronic
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treatments also occurs precociously as noticed by COJOCEL et a1.l in the renal cortex of rats treated with 1200 mg/kg per day CPH for 2 days. Interestingly, the variation in glutathione content is opposite
to the one observed by KU015~16117with massive single CPH dosage. This author attributed the glutathione depletion to the nephrotoxic response to the probable lipid peroxidation mechanism, confirmed by
COJOCEL et .I."? Glutathione as well _as glutathione S-transferase are assumed to play a critical role in detoxication mechanism in response to a lipoperoxidative processl8. However, such a mechanism did not seem to take place in our experimental conditions, and this was confirmed
by the lack of variation in glutathione peroxidase activity. With repeated CPH dosage, the increased glutathione content, the synthesis of which is known to increase rapidly after an acute depletion' 9, associated with the two-fold increase in glutathione Stransferase, corresponded to a marked increase in detoxication and strongly suggest an adaptative phenomenon to the cephaloridine nephrotoxicity. Furthermore, this is well correlated with the absence of histological and enzymic renal disturbances.
/iCKNOWLEffiF MENTS
We wish to thank Prof L. Olivier for histological interpretation, Roussel Uclaf Laboratories for a grant to Miss M.F. Olivier and
21 7
EFFECT OF CEPHALORIDINE TREATMENT O N GLUTATHIONE
A.D.R.T.E. (Association pour le Developpement de la Recherche en Toxicologie Experimentale) for financial support.
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2,
