Methotrexate가 白鼠生殖腺과 기타 장기에 미치는 영향

Abstract

[한글]

The Effects of Methotrexate on the Gonads and Other Organs in Rats

Ha Won Choo

Department of Pathology, Yonsei University College of Medicine Seoul, Korea

Directed by Prof, Dong Silk Kim, M.D.

Assoc. Prof. Yoo Bock Lee, M.D.

Since the report of a dramatic effect of folic acid antagonist on acute leukemia

of children in 1948 by Farber et al. methotrexate has been used widely for the

treatment of various types of malignancy. Li et al. (1958), Li et al.(1960).

Hertz(1961, 1963) and Solomon et al,(1967) reported that methotrexate can not only

arrest but also cure choriocarcinoma. In view of the fact that choriocarcinoma is

originated from embryonal tissue element, methotrexate has been tried to treat

tumors of germ cell origin in the testis and ovary(Mackenzie 1966, Solomon et al.

1967).

Mechanism of the action of methotrexate is attributed to its antagonistic effect

against folic acid which is necessary for synthesis of DNA and RNA as well as for

cell maturation(Werkheizer, 1963, Bertino et al. 1964). By virtue of this effect

methotrexate inhibit the growth of any types of cells which grow rapidly, as well

as neoplastic cells. Most frequently and markedly affected non-neoplastic tissues

include hematopoietic tissue, liver and the mucosa of gastrointestinal tract.

The ovary and testis contain relatively rapidly growing tissue elements, such as

germ cells and follicular cell components. Therefore, it is thought that

methotrexate may affect these continuously growing tissue elements in the gonads.

Present investigation is designed to investigate the effects of methotrexate on the

gonadal tissue in comparison with the previously proved effects on other tissues.

Materials and Methods

Albino rats; young and adult females, young and adult makes, were divided into

four major groups and treated as follows.

Group Ⅰ : Young female rats

A. Normal untreated control(12 rats)

B. Methotrexate administration for 1 course(12 rats)

C. Methotrexate administration for 2 courses(12 rats)

Group Ⅱ : Adult female rats

A. Normal untreated control(12 rats)

B. Methotrexate administration for 1 course(12 rats)

C. Methotrexate administration for 2 courses(12 rats)

Group Ⅲ : Young make rats

A. Normal untreated control(12 rats)

B. Methotrexate administration for 1 course(12 rats)

C. Methotrexate administration for 2 courses(12 rats)

Group Ⅳ : Adult make rats

A. Normal untreated control(12 rats)

B. Methotrexate administration for 1 course(12 rats)

C. Methotrexate administration for 2 courses(12 rats)

Body weight of around 100 gms were used for young group while the body weight of

adult group was around 180 gms. Methotrexate was given orally in a dose of 0.4mg

per kg of body weight once a day for 5 consecutive days for 1 course, and the 2nd

course of 5 days with same dose are given 20 days after the end of the 1st course.

From each group, 4 animals were killed at the 1st, 10th and 20th day after the

ending of methotrexate administration. The changes of body and organ weight

following the treatment of methotrexate were checked, and histological examinations

of the ovary, testes, liver, bone marrow, spleen, and other organs are made.

Tissues for histologic examinations were fixed in 10% neutral formalin, and cut in

6μ. thickness after paraffin embedding, and stained with hematoxylin and eosin.

Results and Summary

There was no notable changes of body weight increase in young animal group, but

the adult groups showed slight depression on the body weight increase after the 2nd

course of methotrexate administration. The weight of ovary and testis did not alter

notably in all groups, but the weight of the liver and spleen showed marked

fluctuation following the administration of methotrexate in all groups. Immediately

after the methotrexate administration, the weight of the liver and spleen

diminished, and it was followed by rapid marked rebound increase at the 10th day

after the cessation of methotrexate treatment, thereafter returning to normal

level.

Histologic alterations in the ovary consisted of maturation arrest of the

developing follicles and depression of luteinization of corpus luteum with

occasional focal necrosis in the corpus luteum, and marked increase of interstitial

cell mass. Histologic alteration of the testis consisted of depression of

spermatogenesis, as evidenced by the decrease of spermatocytes, spermatids and

spermatozoa. Associated with the decrease of cell population, a tendency of nuclear

enlargement was noted, but no multinucleated giant cell formation or frank necrosis

of the cells were found. The most markedly affected cells were spermatocytes and

the least affected were spermatogonia.

The bone marrow showed marked hypoplastc picture with congestion and even

hemorrhage immediately after the cessation of methotrexate treatment, but it

regenerated promptly to normal or even hyperplastic state at the 10th day, The

spleen showed a marked degree of extramedullary hematopoiesis at the 10th day after

the cessation of methotrexate treatment.

Histologic alternations in liver were a mil degree of cloudy welling and fatty

degeneration, and the histologic alterations in the kidrey and adrenals were not

remarkabel.

In summary, administration of methotrexate in a dose fo 0.4mg/kg of body weight

produced rather marked depression on the bone marrow, but induced relatively little

effect on the ovary and testes, indicating that the gonads will not be effected

greatly as long as the dose of methotrexate is tolerable o the bone marrow.

[영문]

Since the report of a dramatic effect of folic acid antagonist on acute leukemia of children in 1948 by Farber et al. methotrexate has been used widely for the treatment of various types of malignancy. Li et al. (1958), Li et al.(1960).

Hertz(1961, 1963) and Solomon et al,(1967) reported that methotrexate can not only arrest but also cure choriocarcinoma. In view of the fact that choriocarcinoma is originated from embryonal tissue element, methotrexate has been tried to treat

tumors of germ cell origin in the testis and ovary(Mackenzie 1966, Solomon et al.1967).

Mechanism of the action of methotrexate is attributed to its antagonistic effect against folic acid which is necessary for synthesis of DNA and RNA as well as for cell maturation(Werkheizer, 1963, Bertino et al. 1964). By virtue of this effect methotrexate inhibit the growth of any types of cells which grow rapidly, as well as neoplastic cells. Most frequently and markedly affected non-neoplastic tissues include hematopoietic tissue, liver and the mucosa of gastrointestinal tract.

The ovary and testis contain relatively rapidly growing tissue elements, such as germ cells and follicular cell components. Therefore, it is thought that methotrexate may affect these continuously growing tissue elements in the gonads.

Present investigation is designed to investigate the effects of methotrexate on the gonadal tissue in comparison with the previously proved effects on other tissues.

Materials and Methods

Albino rats; young and adult females, young and adult makes, were divided into four major groups and treated as follows.

Group Ⅰ : Young female rats

A. Normal untreated control(12 rats)

B. Methotrexate administration for 1 course(12 rats)

C. Methotrexate administration for 2 courses(12 rats)

Group Ⅱ : Adult female rats

A. Normal untreated control(12 rats)

B. Methotrexate administration for 1 course(12 rats)

C. Methotrexate administration for 2 courses(12 rats)

Group Ⅲ : Young make rats

A. Normal untreated control(12 rats)

B. Methotrexate administration for 1 course(12 rats)

C. Methotrexate administration for 2 courses(12 rats)

Group Ⅳ : Adult make rats

A. Normal untreated control(12 rats)

B. Methotrexate administration for 1 course(12 rats)

C. Methotrexate administration for 2 courses(12 rats)

Body weight of around 100 gms were used for young group while the body weight of adult group was around 180 gms. Methotrexate was given orally in a dose of 0.4mg per kg of body weight once a day for 5 consecutive days for 1 course, and the 2nd course of 5 days with same dose are given 20 days after the end of the 1st course.

From each group, 4 animals were killed at the 1st, 10th and 20th day after the ending of methotrexate administration. The changes of body and organ weight following the treatment of methotrexate were checked, and histological examinations

of the ovary, testes, liver, bone marrow, spleen, and other organs are made.

Tissues for histologic examinations were fixed in 10% neutral formalin, and cut in 6μ. thickness after paraffin embedding, and stained with hematoxylin and eosin.

Results and Summary

There was no notable changes of body weight increase in young animal group, but the adult groups showed slight depression on the body weight increase after the 2nd course of methotrexate administration. The weight of ovary and testis did not alter

notably in all groups, but the weight of the liver and spleen showed marked fluctuation following the administration of methotrexate in all groups. Immediately after the methotrexate administration, the weight of the liver and spleen

diminished, and it was followed by rapid marked rebound increase at the 10th day after the cessation of methotrexate treatment, thereafter returning to normal level.

Histologic alterations in the ovary consisted of maturation arrest of the developing follicles and depression of luteinization of corpus luteum with occasional focal necrosis in the corpus luteum, and marked increase of interstitial

cell mass. Histologic alteration of the testis consisted of depression of spermatogenesis, as evidenced by the decrease of spermatocytes, spermatids and spermatozoa. Associated with the decrease of cell population, a tendency of nuclear

enlargement was noted, but no multinucleated giant cell formation or frank necrosis of the cells were found. The most markedly affected cells were spermatocytes and the least affected were spermatogonia.

The bone marrow showed marked hypoplastc picture with congestion and even hemorrhage immediately after the cessation of methotrexate treatment, but it regenerated promptly to normal or even hyperplastic state at the 10th day, The spleen showed a marked degree of extramedullary hematopoiesis at the 10th day after the cessation of methotrexate treatment.

Histologic alternations in liver were a mil degree of cloudy welling and fatty degeneration, and the histologic alterations in the kidrey and adrenals were not remarkabel.

In summary, administration of methotrexate in a dose fo 0.4mg/kg of body weight produced rather marked depression on the bone marrow, but induced relatively little effect on the ovary and testes, indicating that the gonads will not be effected

greatly as long as the dose of methotrexate is tolerable o the bone marrow.