193
PROGESTERONE IV. IN
IN
DEPENDENCE OF THE IN THE RAT UTERUS ON THE Dieter
Egert,
THE
VITRO PROGESTERONE METABOLISM PROGESTERONE CONCENTRATION.
Walter
Jonat
und
Department
of Obstetrics University of Hamburg-Eppendorf,
Received:
UTERUS.
Heinrich
Maass
and Gynaecology Hamburg Germany (FRG)
5/19/T? ABSTRACT
After incubat'on of uterine segments of normal rats s H-progesterone with various concentrations in nutrient medium, different patterns of radioactive steroids were obtained in uterine issue. Using hormone concentrations -4 M progesterone metabolites could of less than 5 x 10 not be detected in the tissue. A series of metabolites appeared with progesterone concentrations of 10w6M and Six radiometabolites were identified and two higher. were characterized. INTRODUCTION Previously
we
progesterone uterine 01 with
and low
The
discrepancy
ly
with
with
of
respect
to
2
other
the
S
amounts
TDBOXDl
of
data
studies
did
possibly
conditions applied
rat
( k-9).
authors
experimental
in
incubation
These
other
detect
progesterone
after
of of
to
labelled
(3).
results
different
able
amounts
segments
investigations to
not
significant
concentrations
by
26, iiwnber
are
injection uterine
hormone
agree
explained
in
we in
after
also
not
is
that
metabolites tissue
2)
Volwne
reported
main-
progesterone.
August,
1975
In
order
study tion
to
the to
check
in
various
vitro
this
possibility
progesterone hormone
MATERIAL
we started metabolism
to in
correla-
concentrations.
AND METHODS
Radioactive steroid. l,2-3H-progesterone (S.A. = 1 Ci/mM) obtained from the Radiochemical Centre Amersham was used for all incubation assays. Before used, the labelled hormone was purified 3 times by preparative TLC (thickness of the layer = 1 mm). The first purification was performed on silica gel HP 2~$sw~;;o;~;z"ne: ethanol 95:5. Unlabelled progesterone graphed alongside. After developing twice, the fraction of labelled progesterone was scraped off the plate, eluted from silica gel with chloroform, and re-chromatographed after evaporation of the solvent. The second purification was performed on silica gel with chloroform : ethanol 95:5 and the third with benzene : ethanol 95:5, developing twice. Reference steroids. Ramat-Gan, Israel: Ikapharm, 3a-Hydroxy-5a-pregnan20-one, 3S-hydroxy-5a-pregnan-20-one, 20a-hydroxy-4pregnen-3-one, 20I3-hydroxy-4-pregnen-J-one, 5a-pregnane3a,20a-dial, 5a-pregnane-30,20a-dial, 5a-pregnane-3a-, 200-diol, and 5B-pregnane-3a,20B-diol. Mann Research Laboratories, New York, USA: 5a-Pregnane3,20-
PROGESTERONE IV. IN
IN
DEPENDENCE OF THE IN THE RAT UTERUS ON THE Dieter
Egert,
THE
VITRO PROGESTERONE METABOLISM PROGESTERONE CONCENTRATION.
Walter
Jonat
und
Department
of Obstetrics University of Hamburg-Eppendorf,
Received:
UTERUS.
Heinrich
Maass
and Gynaecology Hamburg Germany (FRG)
5/19/T? ABSTRACT
After incubat'on of uterine segments of normal rats s H-progesterone with various concentrations in nutrient medium, different patterns of radioactive steroids were obtained in uterine issue. Using hormone concentrations -4 M progesterone metabolites could of less than 5 x 10 not be detected in the tissue. A series of metabolites appeared with progesterone concentrations of 10w6M and Six radiometabolites were identified and two higher. were characterized. INTRODUCTION Previously
we
progesterone uterine 01 with
and low
The
discrepancy
ly
with
with
of
respect
to
2
other
the
S
amounts
TDBOXDl
of
data
studies
did
possibly
conditions applied
rat
( k-9).
authors
experimental
in
incubation
These
other
detect
progesterone
after
of of
to
labelled
(3).
results
different
able
amounts
segments
investigations to
not
significant
concentrations
by
26, iiwnber
are
injection uterine
hormone
agree
explained
in
we in
after
also
not
is
that
metabolites tissue
2)
Volwne
reported
main-
progesterone.
August,
1975
In
order
study tion
to
the to
check
in
various
vitro
this
possibility
progesterone hormone
MATERIAL
we started metabolism
to in
correla-
concentrations.
AND METHODS
Radioactive steroid. l,2-3H-progesterone (S.A. = 1 Ci/mM) obtained from the Radiochemical Centre Amersham was used for all incubation assays. Before used, the labelled hormone was purified 3 times by preparative TLC (thickness of the layer = 1 mm). The first purification was performed on silica gel HP 2~$sw~;;o;~;z"ne: ethanol 95:5. Unlabelled progesterone graphed alongside. After developing twice, the fraction of labelled progesterone was scraped off the plate, eluted from silica gel with chloroform, and re-chromatographed after evaporation of the solvent. The second purification was performed on silica gel with chloroform : ethanol 95:5 and the third with benzene : ethanol 95:5, developing twice. Reference steroids. Ramat-Gan, Israel: Ikapharm, 3a-Hydroxy-5a-pregnan20-one, 3S-hydroxy-5a-pregnan-20-one, 20a-hydroxy-4pregnen-3-one, 20I3-hydroxy-4-pregnen-J-one, 5a-pregnane3a,20a-dial, 5a-pregnane-30,20a-dial, 5a-pregnane-3a-, 200-diol, and 5B-pregnane-3a,20B-diol. Mann Research Laboratories, New York, USA: 5a-Pregnane3,20-
