Isonicotinic Acid Hydrazide 및 Cycloserine의 장기투여가 백서 각장기에 미치는 형태학적 연구

Abstract

[한글]

[영문]

Although among the various antituberculosis drugs, Isonicotinic. Acid Hydrazide(isoniazid) is an excellent drug for the treatment of tuberculosis, in some patients, it may cause toxic reactions. The first, more common type of reaction is seen in the nervous system and is manifested by peripheral neuritis.

This neurotoxicity varies in incidence and severity with the dosage of the drug. The second, less common toxic reactions are thought to be allergic in nature. The original reports on isoniazid described th occurrence of toxic manifestations in

animals and in man(Benson et al., 1952; Bernstein et al., 1952; Elmendorf et al., 1952; Rubin et al., 1952; Steenken & Wolinsky, 1952). After receiving toxic doses animals showed signs of central nervous system stimulation followed by clonic and tonic convulsions, and death resulted from respiratory failure. Different animal species vary considerably in their sensitivity to isoniazid. Clinically the principal form of toxicity is peripheral neuritis, as first reported by Biehl and Skavlem(1953) and Biehl and Nimits(1954). No toxic effects on the peripheral blood or on kidneys have been reported, although a mild transient liver dysfunction was observed. Epileptic patients would seem to be particularly susceptible to the convulsant toxic effect of isomiazid, and fatality in such a patient was reported(Fetterhoff et al., 1952).

The symptoms of isonizid toxicity resemble those of vitamin B^^6 deficiency, and Biehl and Bilter*1954) showed that patients receiving large doses of isoniazid excreted a large amount of pyridoxine, and they found that the complication of peripheral neuritis could be prevented by administering pyridoxine along with isoniazid. In common with the convulsions of B^^6 deficiency, hydrazide induced convulsions can be suppressed immediately by pyridoxine or its congeners (Balzer et al., 1960; Jenney et al., 1953; Ross, 1958). Moreover, pyridoxine does not seem to interfere with the antiacterial action of isoniazid (Crowle & Riemensinder, 1960; Grunberg & BLencowe, 1955; Ungar et al., 1954). The mode of action of isoniazid in producing peripheral neuropathy is not fully understood, but probably isoniazid competes with pyridoxal phosphate for the enzyme, apotrytophanase (Boone & Woodward 1953; Lichstein, 1955), and disturbs the metabolism of the neurone.

Neurological symptoms which are seen frequently after therapeutic dosage, are among the most important of the toxic effects of cycloserine. Also cycloserine produced convulsive seizures and ataxia in mice and rabbits (Fust, 1958). The toxic

action of this tuberculostatic drug, cycloserine, can be explained, at least in part, by its inhibition of B^^6-catalyzed enzymatic reactions(Epstein et al., 1959). Pyridoxine reduces also cycloserine toxicity (Epstein et al., 1959; Robitz다 & Nenashev, 1957). The incidence of acute toxicity following the administration of cycloserine is low, and continued studies in rats, cats, dogs and monkeys showed no toxic effects in theprolonged drug administration except for a temporary anemia and reticulocytosis in dogs and monkeys (Anderson et at., 1956)

Most studies of the toxic action of isoniazid and cycloserine have dealt mainly with clinical, biochemical and antibacterial changes, there are only a few reports of the morphological changes in various organs following prolonged administration

of isoniazid and cycloserine. These change are confined to the nervous system, and simple description of liver, kidney and bone narrow in rats and dogs. Moreover, the aechanism involve in the metabolic and biochemical relationship between isoniazid and cycloserine in large and prolonged doses, and pyridoxine deficiency is poorly understood and still obscure. Very little attention had been paid to the mechanism for the development of organic lesions produced by chronic toxicity of sioniazid

and cycloserine.

Using different experimental conditions, therefore, a detailed morphological study of the lesions of the various organs was made. And special emphasis was made on the effects of prolonged administration of isoniazid and cycloserine-which are known to be related particularly to pyridoxine metabolism. Furthermore, there is still very little known about the mode of action of the factors which cause the organic lesions after prolonged administration of these antituberculosis drugs in rats.

The purpose of this study is to characterize the patterns and define the nature of the morphological changes in various organs in rats, after prolonged administraion of isoniazid and cycloserine. It seeks to clarify the effects of isoniazid administration, the effect of cycloserine administration, their

relationship, and moreover, the effect of the additional administration of synthetic pyridoxine hydrochloride.

Materials and Methods

Healthy male albino rats weighing around 200 Grams were used. The animals were divided into 4 group: Group Ⅰ consisted of 5 rats and normal control; Group Ⅱ consisted of 10 rats receiving isoniazid alone; Group Ⅲ consisted of 12 rats receiving cycloserine alone; Group Ⅳ consisted of 12 rats receiving isoniazed and pyridoxine hydrochlored.

Isoniazid was given by intramuscular injection daily in a dosage of 0.06mg per gm body weight as a 4% solution throughout the experimental period. The dose of cycloserine was 0.09mg per gm body weight per day given by intramuscular injection.

Pyridoxine gydrochloride was given in 0.1mg doses per gm body weight of 5% solution by intramyscular injection daily throughout the experimental period. During the experiment, animal behavior was observed, and body weight was measured weekly.

The animals were sacrificed at the end of 26 weeks, and the weight of the liver was measured. The selected organs were inspected grossly and examined microsceopically. After fixation in 10% formalin, tissue paraffin sections were prepared for hematoxylin-eosin stain, van Gieson stain, Weiger's Resorcin-Fuchsin elastic stain, Gridley's reticulum stain, and modification of Mallory's Reaction for iron. And oil red O staining was performed after frozen section were done.

Results

In the skin, there was some hyperkeratisis and acanthosis associated with a mild degree of edema and cellular infiltration in the corium, but there were no obvious alterations in the appendages, or vascular channels, and no erythema, nor ulceration, in Group Ⅱ. In group Ⅲ, these changes appeared to be less marked. In group Ⅳ, the hyperkeratosis, acanthosis, edema and cellular in filtration were present but to a lesser degree than that seen in group Ⅱ.

In the thyroid gland, the follicles were increased in number. Most of the follicle epithelium was flattened, although some cells were low cuboidal in group Ⅱ. The colloid was more intense in P.A.S stain than that of group Ⅰ. In goup Ⅲ, the follicles were somewhat increased in number, and most of the follicle

epithelium was low cuboidal, but occasionally flat and rarely high cuboidal. The collid was also more intense than that of group Ⅰ. In group Ⅳ. the follicle epithelium was generally low cuboidal, occasionally high cuboidal and rarely flat.

The intensity of colloid were similar to that of group Ⅱ.

In the liver, the lobular arrangement in group Ⅱ was generally similar to that of group Ⅰ. In many areas the liver cells showed severe granular degeneration accompanied by minute areas of necrosis, and rarely vacuolation of the hepatic cells was observed. No fibrosis, hemorrhage, nor changes of bile ducts were

detected. Fat staining of the tissue showed a diffusely granular sudanophilic material located in the centrolobular and mid zonal regions particularly beneath the cell membrane. P.A. S. and reticulum fiber stains showed no particular changes. In group Ⅲ, the granular degeneration of the hepatic cells was less than that of group Ⅱ, and was particularly prominent in the subcapsular region. The sudanophilic material was similar to that of group Ⅱ. In group Ⅳ, the granular degeneration was definitely less than that of group Ⅱ, and in some cases this degeneration was seen only rarely throughout the psecimen. The sudanphilic material was also generally reduced.

In the spleen, is on stain of group Ⅱ showed relative increment of iron pigments, especially located in the red pulp, trabeculae, capsule, perifollicular and perivascular regions, and around the germinal center of splenic follicles. In group Ⅲ, the amount of iron pigment deposited was fenerally similar to that of

group Ⅱ. But in group Ⅳ, iron pigments were markedly reduced.

The adrenal glands appeared to be hypertrophic, particularly in the zona fasciculata which was more marked in group Ⅳ, but there were no obbious alterations in the zone alomerulasa, & zona reticularis, nor in the adrenal meculla.

In the kidneys, there was a similar mild granular degeneration of the renal tubules among group Ⅱ,Ⅲ & Ⅳ.

In the heart, generally no particular changes such as vacuolation, necrosis, hemorrhage, mural thrombi in the atrium fatty infiltration in the myocardium, myocarditis and the vascular changes were observed in all groups. But in group Ⅳ,

the walls of coronary artery sere definitely hypertrophic in two cases and showed a hyaline-like material one of the two. The elastic fiber stain of these two cases showed a beautiful intimal proliferation. Fat staining showed to fatty infiltration in the vascular wall, nor in the myocardium.

Conclusions

The prolonged administration of isoniazid with or without combination of pyridoxine hyerochloride, and cycloserine in large doses in rats induced morphological changes in the skin, liver, thyroid glands, kidney and coronary arteries.

1. The major morphological changes induced by isoniazid and cycloserine are hyperkeratiosis and acanthosis of epidermis, and edema and cellular infiltration of dermis in the skin, granular and fatty degenerations of hepatic cells in the liver, retrogressive changes of follicle in the thyroid gland, hyperttrophy of adrenal cortex, mild granular degeneration of tubules in the kidneys, and increased iron deposition in the spleen.

2. These morphological changes of the organs were somewhat lesser in the group receiving cycloserine only than isoniazid only, and were identical with those induced by pyridoxine deficiency.

3. The morphological changes being induced by isoniazid were minimized in part by the combined administration of pyridoxine hydrochloride, but were not completely prevented.

4. The two cases of arteriosclerotic lesions with intimal proliferation of coronary arteries in the group of combined administration of isoniazid and pyfidoxine are very interesting and equivocal, whether these lesions are relate to spontaneous occurrence, or to the effect of large doses of pyridoxine

hydrochloride, or other unknown factors.

It appears that the mechanism involving morphological changes induced by isoniazid and cycloserine, and involving the partial prevention of the lesions by combined administration of pyridoxine hydrochloride is a complex metabolic derangement and also is difficult to interpret. And the molphological changes

induced by isoniazid and cycloserine appeared not to be due to deficiency of pyridoxine only, but still remains for particular interpretation.