In vitro and in vivo studies on the effect of tolbutamide and phenformin on adrenal steroidogenesis in the rat

Pharmacological

IN

Research

VITRO

AND

Communications,

IN

PHENFORMIN

VIVO

Vol. 3, No. 4. 7971

STUDIES

ON ADRENAL

ON THE

of

Pharmacology,

The

effect

steroidogenesis

Faculty

of

was

of

adrenal

as

a measure

for

of

corticosterone.

of

25,

50 and

100

of

the

enzyme

activity,

corticosterone.

did

not

The

conclusion

exerts

in

the

2.5

mg per

reached

release

adrenal

glands.

of

150

250

and

with

mg/kg

at either

mg per

of

the

tolbutamide

increase

by

in

at

levels doses

a decreased

reveals

decreased

1.25

plasma

inhibition

study

mg adrenal

admini-

caused

phenformin

a significant

vitro

taken

Oral

and

accompanied

In

that

estimations were

in

was

100

The

adrenal

phosphorylase

100

two

that

mg adrenal

parameters.

induces

the

steroidogenesis;

stimulation phenformin

effect.

It

1957

rat.

in

which

opposite

the

phosphorylase

resulted

a subsequent

the

University

on

corticosterone

adrenal

affect is

stimulates ) and

doses

Phenformin

INTRODUCTION

Cairo

phenformin

treatment

mg/kg

significantly

with

Pharmacy,

and

plasma

corticosterone.

at

and

ACTH

at

Similar

plasma

tolbutamide

of

and

increase

of

in

steroidogenesis

of

level

THE RAT

Egypt.

investigated

tolbutamide

a significant

of

tolbutamide

phosphorylase

stration

IN

AND

1971

7 November

SUMMARY

OF TOLBUTAMIDE

El-Denshary

Cairo, Received

EFFECT

STEROIDOGENESIS

E.S.M. Department

355

has

adrenal

been

reported

glands

to

( Hechter

et

that secrete

al.,

corticotropin ( Haynes

1951

) an

(ACTH) and

increased

Berthet, quantity

356

Pharmacological

of

corticosteroid

action by

revealed

an

that

increased

( Haynes could the

hormones.

Berthet,

therefore

be

effect

object of

assessed

adrenal

the

taken

).

as

ACTH

stimulation

The

in amount

a measure

for

of

accompanied

the of

Vol. 3. No. 4. 1971

mechanism

is

activity

1957

by

and

MATERIALS

in

doses

were

treated

divided

of

adrenal

tissue

adrenal

phosphorylase

steroidogenesis

100

and

The

in

and filter

homogenized activity

with in

0.1

the

of

inorganic

was

the

quantity

of

phosphate

the was

the

made

was

groups for

were gland

to

of

and of water

highest

doses

water

and

to the

tubes trimmed

dried of

between

glands

method

a

was

Phosphorylase

according is

glycogen

to

Shinowara

the

based

on

released and

method the from

5-adenosine

phosphorylase. to

groups

doses

isolated,

pair

of

at

Vitamin

in

phosphate

presence

water

administration

solution.

This

received

received

the

was

inorganic

according

at

collected

Each

influence

U.S.

drug

determined

(1956).

150-

Other

access

after

fluoride

Wosilait

in

free

Each

M sodium

weighing

in

water

used

glands

and

under

in

weighed.

homogenate

glucose-l-phosphate, monophosphate,

and

phospho-

Groups

( DBI,

blood

ice.

corti-

adrenal

respectively.

those

adrenal

plasma

) dissolved

allowed

the

under

paper

Sutherland

estimation

The

placed

each.

Control

hours

and

on

rats

10

respectively.

Twenty-four

vivo as

and

rats

mg/kg,

to

in

steroidogenesis

albino

phenformin

were

the

studied.

) dissolved

rats

heparin.

of

250

study

drugs

Hoechst

comparable

decapitated

containing

of

with

mg/kg,

milk.

were

fat

and

both

male

groups

150

to on

also

( Rastinon,

volumes

drugs.

piece

into

is

phosphorylase of

Adult

Corporation

in

from

effect

similarly

50 and

rats

adrenal

was

100,

Pharmaceutical

bread

vitro

work phenformin

of

sodium

oral

the

and

AND METHODS

tolbutamide

orally

present

corticosterone

g were

25,

the

estimation The

rylase

200

of

tolbutamide

costerone.

of

the

on

CommunicationsS

gland. The

of

Studies

phosphorylase

and

Research

Estimation et

al.

of (1942).

Pharmacological

Research

The

results

unit

can

were be

1 mg of

Communications,

expressed

defined

as

inorganic

conversion

of

level

of

were

phosphorous

in

was

Homogenates

of in

were

absence

the

the

of

which ten

fluoride

at

for

tolbutamide

homogenates

(2.5

was

(1955).

100

the

the

range the

Der

12 to

Vies

of

six

control

solution. 30 minutes

measured

of

in

(1960). rats

The

homo-

the

presence

LOO mg adrenal), The

was

or

corticosterone

according

phosphorylase

22%.

corticosterone

Van

mg per

One

percentage

of

to

mg adrenal).

then

Adrenal

in

weight. release

when

and

M sodium

gram the

minutes

glands

30°C

per

causes

according

adrenal

mg per

unit

centrifuged

measured

0.1

(1.25

Schally

of

is

was

incubated

phenformin in

amount

blood

prepared

and

terms

the

plasma

genates

in

glucose-l-phosphate

Heparinized

357

Vol. 3. No. 4, 1971

to

Saffran

estimated

and

as

previously

mentioned.

RESULTS

Tolbutamide

significant

effect

sterone.

Phenformin

a significant plasma

was

proportional

are

given

level

in

in

phenformin

did

doses in

mg/kg

produced

to

the

dose.

250

level of

of

25,

show

Results

adrenal

phosphorylase

enzyme 100

of

obtained

for

corticoproduced

phenformin both

drugs

Fig.1. induced

and

corticosterone.

affect

caused

and

mg/kg

tolbutamide

significantly

drug

activity

effect

in

cortico-

the

phosphorylase The

that

the

no

plasma

mg/kg

50 and

illustrated

results

or

and

adrenal

corticosterone.

Results mg/kg

costerone. the

150

the

of

not

100

the

significant However,

level

of

the

enzyme

hormone.

DISCUSSION 250

of

in

1 and

of

phosphorylase

doses

at

Table

vitro

decreases

and

adrenal

reduction

and

the

at

a dose

elevation

sterone.

or

on

However,

a significant

In

at

adrenal

reveal

elevated

adrenal

Phenformin, enzyme

and

that

tolbutamide

at

phosphorylase

on the

other

hand,

plasma

corticosterone

doses

and was

plasma

found at

of

doses

150 corti-

to

decrease of

25,

50 and

and

control 100 mg/kg 150 mg/kg 250 mg/kg

+

sodium

Dosage

abcd-

2)

control 25 mdkg 50 mg/kg 100 mg/kg

Phenformin

*

----------------.---------_-----me----------

abcd-

.)Tolbutamide

Drug

1

26.60 19.80 15.21 11.50

33.70 35.34 38.18 37.20

Units adrenal

drug at

2 2 2 +

2 : t 2

100.0 74.43 57.14 43.23

100.0 106.05 113.30 110.30

Percentage to control

Phosphorylase

was given orally. doses comparable

0.90 1.80 1.50 1.01

1.07 1.55 1.58 0.96

per g ? S.E.

Adrenal

to

The rats those

P = 0.01 P =O.OOl P =O.OOl

P = 0.5 P = 0.05 P = 0.05

Significance

of the used for

16.31 12.21 9.02 8.61

15.82 16.73 19.61 18.81

~g per plasma

1.20 1.13 0.87 0.88

1.02 1.07 1.04 1.11

received doses of

100.0 74.86 55.30 46.65

100.0 105.75 120.10 118.90

Percentage to control

Corticosterone

water the

P = 0.05 P = 0.00 P = 0.00

P = 0.5 P = 0.05 P = 0.05

Significance

and plasma level of hours after adminimade on adrenal homoge-

control group the highest

+ ? + ”

2 + 'r 2

100 ml i S.E.

Plasma

sodium and phenformin on rat adrenal phosphorylase Twenty-four each consisting of 10 rats, were used. the rats were sacrificed and the estimations were

Each orally drugs.

Effect of tolbutamide corticosterone. Groups, stration of the drugs, nate and plasma.

TABLE

Pharmacological

Research

Communications,

ToL

8 U TAHIDE

Effect of tolbutamide and plasma corticosterone. The drugs were given after administration.

PHENFORMIN

and

phenformin on rat Groups of 10 rats and the animals

orally

Phosphorylase

I

359

Vol. 3, No. 4, 1971

unit

Corticosterone

per

(mcg

adrenal phosphorylase were used. each were used 24 hours

g adrenal

per

100

ml

plasma)

lssl Standard

I

100 of

mg, ACTH

Berthet by

Error.

respectively. results

in

(1957)

stimulating

a stimulation

of

the

corticoid

that

ACTH

activation

and

adrenal

promotes

the

results

may

The

by

stimulation Haynes

of

therefore

steroidogenesis

is

by

ACTH

based

causes

adrenal indicate

relationship

phosphorylase enzyme

the

steroidogenesis

The

phenformin.

adrenal of

ACTH

(1955), production.

corticosterone

by and

Vogt

activity.

plasma

inhibition

steroidogenesis that

increased

phosphorylase and

their

to

postulated

phosphorylase

and

an

According

tolbutamide between

on larger

the

fact amounts

and

Pharmacological

360

Research

TABLE In

vitro

effect

of

sodium

and

corticosterone.

1. of

:“c

6

experiments

on

adrenal

Phenformin (1.25 mg per 100 mg adrenal)

6

6

unit 2 S.E.

29*23

+ 2.31 -

16.66

+ 1.47*

28.56

+ 2.17

[renal corticostelne (pg per 100 mg adrenal) ? S.E.

2.91

+ 0.04

1.73

‘r 0.06~

2.89

+ 0*02

kosphorylase g adrenal

3e :r .id

phenformin

Tolbutamide sodium (2.5mg per 100 mg adrenal)

Control

Pt

and

1

c

.k

Vol. 3, No. 4, 1971

2

tolbutamide

phosphorylase

Communications,

t

*

Significant

of

from

glucose.l-phosphate

Berthet, the

1957

to

system

generate

steps

in

the

xylation of

adrenal

cortex

the

gland In

alter

supported

is

accompanied

in

vivo

1955 is

196.1 the

by

).

fact

that

phenformin

and

corticosterone.

stimulant

drug

on

of

levels

drug

effect

is of

in

mediated tolbutamide

dehydrogenase several the

concept

hydroof

stimulation the

the of

glycogen

the

content

). did

not

significantly

This

parameters.

and

adrenal the

1955

both

phosphorylase

their

effect

the

The

by

for

This

in

reveals

of

).

in

that

a fall

and

oxidized

required

study

phosphorylase

( Haynes

particularly

al., by

al.,

Papageorge,

adrenal

decreased

The

is

effect

it

et

largely

which

steroids,

( White

is

and

adrenal

increased

of

p = 0.01.

glycogen

( Noble

vitro

a direct

et

NADP

at

from

( Kelly

synthesis

ACTH

control

formed

reduced

reaction

action

be

the

Glucose-l-phosphate

).

dehydrogenase

reactions

of

decrease

plasma

may

Although

exclude tolbutamide

corticosterone

vitro.

This

through

may ACTH

on ACTH

is

in indicate

vivo, that

the

activation. supported

by

the

Pharmacological

Research

finding

Communications,

reported

depletes

by

adrenal

mulation

of

in

as

it

was

between

( Jollife,

1953

with

the

finding

drug

causes

The of

an

in

the

rat

that

et

that

at

evidenced

of

in

El-Aziz

ascorbic

effect

increase

Abd

ascorbic

Miller

361

adrenal

adrenal

).

4. 1971

and

acid

depletion

ACTH

relationship

El-Denshary

ascorbic

The

250 w/kg.

Vol. 3, No.

doses

plasma

is

an

content

tolbutamide

and

ACTH

reported

corticosteroids

in

sti-

inverse

and

on who

150

indicates

there

a1.(1966)

of

acid

acid

tolbutamide

ACTH

activity

is

consistent

that

the

the

rhesus

monkey. It

was

revealed

by

gluconeogenesis and

from

Williams, the

present

of

steroidogenesis

promote

work

agents

liver

in in

produces

the

adrenal

likely

on

liver

present

stimulation

and to

be

phosphorylase

rylase,

on the glucagon

'The

(Rail

conclusion

adrenal

phenformin

phosphorylase might

and that rat

(Ciron

to

the

inhibition

agent,

increases

(Hildmann

and was

the

found

fact

ACTH and

was

the the

effect

ACTH

of phosphorylase

has

no

Liver

stimulated

phosphoby

epinephrine

1957).

therefore

reached produced through

that by

their

effect

stimulates

1957). be

inhibition

adrenal

specifically

to

On the

tolbutamide

that

Berthet,

found

1965).

that

liver

1958)

phosphorylase

Zillmann,

and

by

al.,

liver

on

drugs

hypoglycemic causes

et

on

be mediated

finding

tolbutamide

controversy

activity

the

biguanide

causes

al.,

to

glucocorticoids

phenformin

hand,

is

that

in

(Haynes

et

attributed

sulfonylurea

However,

other

be

Steiner

1969).

This

phosphorylase.

According

while

biguanide

decreases 1957;

it

interpreted

adrenal

and

may

al.,

work,

phosphorylase.

sulphonylurea

19.58).

activity

guinea-pig

the

et

reveal

enzyme

phenformin

evidenced

of

a biguanide the

contrary,

was

effect

the

buformin,

is

effect

phosphorylase

of

activity

al.,

(Harper,

on the

inhibition

of

this

gluconeogenesis

on

while

et

it

that

(Williams

Wick

as

Studies

authors

protein

1958;

of

many

the

change

tolbutamide effects

in and

on

ACTH.

362

Pharmacological

Research

Communications,

Vol. 3, No.

4. 197 7

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Hyde

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Nielsen

R.L.,

(1957),