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Resistance of the guinea pig to indomethacin ulcerogenesis
TOXICOMGY
AND
APPLIED
PHARMACOLOGY
SHORT Resistance
of the Guinea
45,637-639
(1978)
COMMUNICATIONS Pig to Indomethacin
Ulcerogenesis
Resistance of the Guinea Pig to Indomethacin Ulcerogenesis. MARIANI, L., AND BONANOMI, (1978). Toxicol. AppL Phnrmacol. 45, 637-639. Steroid or nonsteroid anti-inflammatory drugs are said to induce gastrointestinal lesions in laboratory animals and man. Oral or parenteral administration of indomethacin (12 mg/kg) produced a 100% incidence of intestinal ulcerations, peritonitis, and death in the rat. In contrast, indomethacin failed to induce any intestinal or gastric ulceration in the guinea pig after a single intravenous dose (25-50 mg/kg) or prolonged oral administration (50, 100, and 150 mg/kg/day for 15 days). The results show a clear-cut species difference for the toxic effects of indomethacin and cortisone. The relationship, if any, between ulcerogenic sensitivity and the inhibition of indomethacin and cortisone of prostaglandin biosynthesis or release is not known. L.
It is generally accepted that both in man (Liivgren and Allander, 1964); (Sturges and Krone, 1973) and experimental animals (Somogyi et al., 1969; Brodie et al., 1970) prolonged treatment with steroid and nonsteroid anti-inflammatory drugs can induce gastric or intestinal ulcers and can give rise to reactivation or exacerbation of quiescent or latent ulcers. Previous investigations (Heisler and Kovacs, 1969) showed that guinea pigs, in contrast with rats, are insensitive to the ulcerogenic action of cortisone. We have observed that indomethacin, even at high doses, failed to develop ulcers in guinea pigs and have now administered multiple oral doses and single iv doses of indomethacin to guinea pigs. For comparison, a similar experiment was conducted in the rat, since it is well known that in this species oral or parenteral administration of indomethacin in single large doses, or multiple smaller doses, produces a 100% incidence of peritonitis and death (Brodie et al., 1970). METHODS
Studies were performed on Pirbright male guinea pigs (400-500 g) and male albino rats of the Wistar-Charles River strain (200-250 g). The animals were fed laboratory chow (Altromin) and water ad libitum. Preliminary acute toxicity experiments were performed in both guinea pigs and rats to determine the oral LD50 of indomethacin. In these experiments, the animals were observed for 7 days after drug administration, and the LD50 values were calculated by the method of Lit&held and Wilcoxon (1949). For ulcerogenic effects, guinea pigs, randomized into groups of 10 animals were housed, 2 animals per cage, in wire-bottom cages to minimize coprophagy and administered indomethacin orally (0, 50, 100, or 150 mg/kg/day, for 15 days) in an aqueous suspension of 1% carboxymethylcellulose at a volume of 5 ml/kg; controls received the vehicle only. Additional groups of guinea pigs were treated intravenously 637
0041-008x/78/0452-0637$02.00/0 Copyright Q 1978 by Academic Press, Inc. All rights of reproduction in any form resewed. Printed in Great Britain
SHORT
638
COMMUNICATIONS
after light ether anesthesia. The animals received indomethacin solubilized in a 0.1 M phosphate buffer at pH 8.1 at a volume of 1 ml/kg. Rats were randomly distributed into groups of 10 animals and administered the drug as indicated in Table 1. At the end of the selected treatment period (24 hr after the last repetitive dose or 72 hr after the single TABLE
I
THE ULCEROGENIC EFFECTS OF A SINGLE DOSE OF INDOMETHACIN IN RATV
Gastrointestinal ulcers Group 1 2 3
Stomach
Small intestine
Large intestine
Number that died
12po
0 0
65 100
0 10
o/10 3110
12iv
0
100
0
l/10
Indomethacin bdkd 6~0
a Ten rats were used in each group.
dose) the animals were killed. Stomach and intestines were removed and the incidence of ulcerative lesions was determined. RESULTS AND DISCUSSION The oral LD50 values of indomethacin in rats and guinea pigs were found to be 38 (29-46) and 480 (360-600) mg/kg, respectively. In the guinea pig, indomethacin administered orally (SO-lQ0 and 150 mg/kg/day) for 15 days or iv in a single dose (25-50 mg/kg) did not induce gastric or intestinal lesions. In the rat, a single dose of 12 mg/kg indomethacin given orally or iv produced intestinal ulcers in all of the animals and a high incidence of perforations. The lesions were found almost exclusively in the midportion of the small intestine. Occasionally, ulcers were also found in the large intestine (Table 1). From the results of this investigation it appears that the guinea pig, in contrast with the rat, is insensitive to the ulcerogenic action of indomethacin. Intestinal lesions occurred in all rats given a single dose of the drug, whereas in the guinea pig, indomethacin given at higher doses and on a multiple dose schedule failed to induce lesions. In addition, the LD50 of indomethacin in rats was 10 to 15 times lower than that in the guinea pig, showing a relationship between &erogenic and toxic effects in these two species. Heisler and Kovacs (1969) reported a difference in the ulcerogenic action of cortisone in these two species. In their studies, in contrast with rats, guinea pigs appeared insensitive to cortisone, which Heisler and Kovacs postulated was due to a protective factor present in the guinea pig but not in the rat. A similar protective fact, if it exists, might also be involved with respect to indomethacin. It is not known if sensitivity to the ulcerogenic activity of nonsteroid and steroid antiinflammatory drugs may correlate primarily with a difference in the structure of the intestines, with metabolism, with enterohepatic circulation of these drugs (Klaassen,
SHORT
COMMUNICATIONS
639
1976), or with the known capacity of these drugs to inhibit biosynthesis or release of prostaglandins (Ferreira et al., 1971; Flower, 1974; Vane, 1971; Lewis and Piper, 1975). REFERENCES D. A., COOK, P. G., BAUER, B. J., AND DAGLE, G. E. (1970).Indomethacin-induced intestinallesionsin the rat. Toxicol. Appl. Pharmacol.17,6 15-624. FERREIRA, S. H., MONCADA, S., AND VANE, J. R. (1971). Indomethacinand aspirinabolish prostaglandinreleasefrom the spleen.Nature NewBiol. 231,237-239. FLOWER, R. J. (1974).Drugs which inhibit prostaglandinbiosynthesis. Pharmacol.Rev. 26,33BRODIE,
67. S., AND KOVACS, E. M. (1969). The resistanceof guinea pigs to cortisone ulcerogenesis. J. Pharm.Pharmacol.21,269-270. KLAASSEN, C. D. (1976). Studieson the mechanismof spironolactoneprotection against indomethacintoxicity. Toxicol. Appl. Pharmacol.38, 127-135. LEWIS, G. P.,AND PIPER, P. T. (1975).Inhibition of prostaglandinrelease by anti-inflammatory steroids.International Conferenceon Prostaglandins. Florence,1975(Abstract). LITCHFIELD, J. T., AND WILCOXON, F. (1949). A simplifiedmethodof evaluatingdose-effect experiments.J. Pharmacol.Exp. Ther. 96,99-l 13. LGVGREN, O., AND ALLANDER, E. (1964).Sideeffectsof indomethacin.Brit. Med. J. 1, 118. SOMOGYI, A., KOVACS, K., AND SELYE, H. (1969).Jejunalulcersproducedby indomethacin.J. Pharm.Pharmacol.21,122-123. STURGES, H. F., AND KRONE, C. L. (1973).Ulceration andstructureof the jejunumin a patient on long-termindomethacintherapy. Amer. J. Gastroenterol.59, 162-169. VANE, J. R. (1971). Inhibition of prostaglandinsynthesisas a mechanism of action for aspirinlike drugs.Nature NewBiol. 231,232-235. HEISLER,
L. MARIANI L. BONANOMI
Departmentof Pharmacologyand Therapeutics University of Milan Milan, Italy ReceivedMay 13,1977; acceptedMarch 9,1978
AND
APPLIED
PHARMACOLOGY
SHORT Resistance
of the Guinea
45,637-639
(1978)
COMMUNICATIONS Pig to Indomethacin
Ulcerogenesis
Resistance of the Guinea Pig to Indomethacin Ulcerogenesis. MARIANI, L., AND BONANOMI, (1978). Toxicol. AppL Phnrmacol. 45, 637-639. Steroid or nonsteroid anti-inflammatory drugs are said to induce gastrointestinal lesions in laboratory animals and man. Oral or parenteral administration of indomethacin (12 mg/kg) produced a 100% incidence of intestinal ulcerations, peritonitis, and death in the rat. In contrast, indomethacin failed to induce any intestinal or gastric ulceration in the guinea pig after a single intravenous dose (25-50 mg/kg) or prolonged oral administration (50, 100, and 150 mg/kg/day for 15 days). The results show a clear-cut species difference for the toxic effects of indomethacin and cortisone. The relationship, if any, between ulcerogenic sensitivity and the inhibition of indomethacin and cortisone of prostaglandin biosynthesis or release is not known. L.
It is generally accepted that both in man (Liivgren and Allander, 1964); (Sturges and Krone, 1973) and experimental animals (Somogyi et al., 1969; Brodie et al., 1970) prolonged treatment with steroid and nonsteroid anti-inflammatory drugs can induce gastric or intestinal ulcers and can give rise to reactivation or exacerbation of quiescent or latent ulcers. Previous investigations (Heisler and Kovacs, 1969) showed that guinea pigs, in contrast with rats, are insensitive to the ulcerogenic action of cortisone. We have observed that indomethacin, even at high doses, failed to develop ulcers in guinea pigs and have now administered multiple oral doses and single iv doses of indomethacin to guinea pigs. For comparison, a similar experiment was conducted in the rat, since it is well known that in this species oral or parenteral administration of indomethacin in single large doses, or multiple smaller doses, produces a 100% incidence of peritonitis and death (Brodie et al., 1970). METHODS
Studies were performed on Pirbright male guinea pigs (400-500 g) and male albino rats of the Wistar-Charles River strain (200-250 g). The animals were fed laboratory chow (Altromin) and water ad libitum. Preliminary acute toxicity experiments were performed in both guinea pigs and rats to determine the oral LD50 of indomethacin. In these experiments, the animals were observed for 7 days after drug administration, and the LD50 values were calculated by the method of Lit&held and Wilcoxon (1949). For ulcerogenic effects, guinea pigs, randomized into groups of 10 animals were housed, 2 animals per cage, in wire-bottom cages to minimize coprophagy and administered indomethacin orally (0, 50, 100, or 150 mg/kg/day, for 15 days) in an aqueous suspension of 1% carboxymethylcellulose at a volume of 5 ml/kg; controls received the vehicle only. Additional groups of guinea pigs were treated intravenously 637
0041-008x/78/0452-0637$02.00/0 Copyright Q 1978 by Academic Press, Inc. All rights of reproduction in any form resewed. Printed in Great Britain
SHORT
638
COMMUNICATIONS
after light ether anesthesia. The animals received indomethacin solubilized in a 0.1 M phosphate buffer at pH 8.1 at a volume of 1 ml/kg. Rats were randomly distributed into groups of 10 animals and administered the drug as indicated in Table 1. At the end of the selected treatment period (24 hr after the last repetitive dose or 72 hr after the single TABLE
I
THE ULCEROGENIC EFFECTS OF A SINGLE DOSE OF INDOMETHACIN IN RATV
Gastrointestinal ulcers Group 1 2 3
Stomach
Small intestine
Large intestine
Number that died
12po
0 0
65 100
0 10
o/10 3110
12iv
0
100
0
l/10
Indomethacin bdkd 6~0
a Ten rats were used in each group.
dose) the animals were killed. Stomach and intestines were removed and the incidence of ulcerative lesions was determined. RESULTS AND DISCUSSION The oral LD50 values of indomethacin in rats and guinea pigs were found to be 38 (29-46) and 480 (360-600) mg/kg, respectively. In the guinea pig, indomethacin administered orally (SO-lQ0 and 150 mg/kg/day) for 15 days or iv in a single dose (25-50 mg/kg) did not induce gastric or intestinal lesions. In the rat, a single dose of 12 mg/kg indomethacin given orally or iv produced intestinal ulcers in all of the animals and a high incidence of perforations. The lesions were found almost exclusively in the midportion of the small intestine. Occasionally, ulcers were also found in the large intestine (Table 1). From the results of this investigation it appears that the guinea pig, in contrast with the rat, is insensitive to the ulcerogenic action of indomethacin. Intestinal lesions occurred in all rats given a single dose of the drug, whereas in the guinea pig, indomethacin given at higher doses and on a multiple dose schedule failed to induce lesions. In addition, the LD50 of indomethacin in rats was 10 to 15 times lower than that in the guinea pig, showing a relationship between &erogenic and toxic effects in these two species. Heisler and Kovacs (1969) reported a difference in the ulcerogenic action of cortisone in these two species. In their studies, in contrast with rats, guinea pigs appeared insensitive to cortisone, which Heisler and Kovacs postulated was due to a protective factor present in the guinea pig but not in the rat. A similar protective fact, if it exists, might also be involved with respect to indomethacin. It is not known if sensitivity to the ulcerogenic activity of nonsteroid and steroid antiinflammatory drugs may correlate primarily with a difference in the structure of the intestines, with metabolism, with enterohepatic circulation of these drugs (Klaassen,
SHORT
COMMUNICATIONS
639
1976), or with the known capacity of these drugs to inhibit biosynthesis or release of prostaglandins (Ferreira et al., 1971; Flower, 1974; Vane, 1971; Lewis and Piper, 1975). REFERENCES D. A., COOK, P. G., BAUER, B. J., AND DAGLE, G. E. (1970).Indomethacin-induced intestinallesionsin the rat. Toxicol. Appl. Pharmacol.17,6 15-624. FERREIRA, S. H., MONCADA, S., AND VANE, J. R. (1971). Indomethacinand aspirinabolish prostaglandinreleasefrom the spleen.Nature NewBiol. 231,237-239. FLOWER, R. J. (1974).Drugs which inhibit prostaglandinbiosynthesis. Pharmacol.Rev. 26,33BRODIE,
67. S., AND KOVACS, E. M. (1969). The resistanceof guinea pigs to cortisone ulcerogenesis. J. Pharm.Pharmacol.21,269-270. KLAASSEN, C. D. (1976). Studieson the mechanismof spironolactoneprotection against indomethacintoxicity. Toxicol. Appl. Pharmacol.38, 127-135. LEWIS, G. P.,AND PIPER, P. T. (1975).Inhibition of prostaglandinrelease by anti-inflammatory steroids.International Conferenceon Prostaglandins. Florence,1975(Abstract). LITCHFIELD, J. T., AND WILCOXON, F. (1949). A simplifiedmethodof evaluatingdose-effect experiments.J. Pharmacol.Exp. Ther. 96,99-l 13. LGVGREN, O., AND ALLANDER, E. (1964).Sideeffectsof indomethacin.Brit. Med. J. 1, 118. SOMOGYI, A., KOVACS, K., AND SELYE, H. (1969).Jejunalulcersproducedby indomethacin.J. Pharm.Pharmacol.21,122-123. STURGES, H. F., AND KRONE, C. L. (1973).Ulceration andstructureof the jejunumin a patient on long-termindomethacintherapy. Amer. J. Gastroenterol.59, 162-169. VANE, J. R. (1971). Inhibition of prostaglandinsynthesisas a mechanism of action for aspirinlike drugs.Nature NewBiol. 231,232-235. HEISLER,
L. MARIANI L. BONANOMI
Departmentof Pharmacologyand Therapeutics University of Milan Milan, Italy ReceivedMay 13,1977; acceptedMarch 9,1978