Merphological Effect of Excess Copper Sulfate on the Adrenal Gland and Anterior
Hypophysis of the Rat
Sang Ho Cho, M.D.
Department of Medical Science, The Graduate School, Yonsei University
(Directed by Prof. Dong Sik Kim, M.D. and Prof. Yoo Bock Lee, M.D.)
The mechanism by which copper exerts its toxic effect on the hepatic cells in
Wilson's disease is not known yet.
Attempts to produce hepatic dysfunction and morphological lesions equivalent to
Wilson's disease in experimental animals have been unsuccessful. Experimentally
induced deposits of copper in hepatic cell generally do not produce necrosis or
cirrhosis (Howell, 1959 ; McNary, 1963; Barka et al., 1964).
Recently, results of many experiments on this subject suggested that
sequestration of copper by lysosomes may protect hepatocytes from toxic effects of
this metal (Goldfischer and Moskal, 1966; Goldfischer, 1967; Goldfischer and
Sternlieb, 1968; Goldfischer et al., 1970). The corticosteroid is known to
stabilize lysosome in vivo (Weissmann, 1965).
According to the results of many experiments, one of the consequences of
adrenalectomy or hypophysectomy in rats is an increased ceruloplasmin with a
restoration of normal levels when the animals are given corticosterone (Evans and
Wiederanders, 1968; Gregoriades and Sources, 1969: Evans et al., 1970). On the
basis of these results, Evans and Cornatzer (1971) have considered that following
adrenalectomy or hypophysectomy, bile flow was significantly decreased suggesting
that adrenal steroids regulate biliary copper excretion by choleretic action.
But studies of the role of the adrenals in the metabolism of copper have been
concerned mainly with effect of adrenalectomy and of certain corticosteroids on the
activity of plasma ceruloplasmin (El-Mofty et al., 1959; Evans and Wiederanders,
Attempting to gain a better understanding of the role of the adrenals in copper
metabolism, the influence of administration of excess copper on both adrenals and
anterior hypophysis is examined.
Materials and Methods
Adult female albino rats were divided into two groups and treated as follows:
Group Ⅰ: Normal control 24 rats
Group Ⅱ: CuSO^^4 treated 84 rats
In experimental group, 0.3 c.c. of 0.5% copper sulfate solution was given
intraperitoneally per animal daily and 0.3 c.c. of distilled water was given by
same method in control group. At 1st, 3rd, 5th, 7th, l0th, 15th, and 30th days, 12
rats were killed in experimental group and 6 rats in control group by
The unfixed adrenals and pituitary glands were weighed on the chemical balance in
order to verify enlargement.
For light microscopic examination, adrenals and liver were fixed in 10% neutral
formalin and pituitary glands were fixed in Zenker's solution. They were embedded
in paraffin and stained with hematoxyli-eosin. Additionally, liver sections stained
with diethyldithiocarbamate stain (Howell, 1959) and pituitary sections stained
with P.A.S. reaction. Sudan stain with the adrenal sections were performed.
Specimens for Schultz reaction were obtained from the adrenal gland. The
specimens were fixed in formol-calcium for 2 to 3 days and cut 10μ frozen
sections. And then they were washed for 24 hours in several changes of distilled
water, placed he sections into ferric ammonium sulfate solution for one week at
37℃ washed sections in 3 changes of acetate buffer one hour in each, rinsed
quickly in distilled water, placed in 5% formalin for 10 minutes, mounted on
slides, applied a drop of acetic-sulfuric acid mixture (1:1) to a cover slip and
examine immediately under the microscope.
Results and Summary
1) The intracellular deposition of copper was increased gradually, but neither
hepatic cell necrosis nor cirrhosis were noted throughout the whole experimental
2) The excess amount of copper sulfate loading caused increased weight and
hyperplastic changes of zona fasciculata and zona reticularis combined with
increased accmulation of lipids in adrenal gland of adult rat after the
administration as early as the first day. And also increased functional activity
was induced. However, these hyperplastic changes and increased functional activity
in adrenal gland did not progressively increase as copper loading increased.
3) In anterior hypophysis, increased weight and hyperplastic changes of beta
cells were caused after the administration of the first day. But these changes did
not progressively increase as copper loading increased.
On the basis of the above results it is speculated that an increased amount of
adrenocortical hormones as a result of adrenocortical hyperfunction, mediated by
the pituitary, in excess copper loaded rats may protect the hepatic cells from the
toxic effects of excess copper.