BIOLOGICAL-ACTIVITIES OF THE INTESTINAL MICROFLORA IN MICE TREATED WITH ANTIBIOTICS OR UNTREATED AND THE EFFECTS OF THE MICROFLORA ON ABSORPTION AND METABOLIC-ACTIVATION OF ORALLY-ADMINISTERED GLUTATHIONE CONJUGATES OF K-REGION EPOXIDES OF 1-NITROPYRENE
T. Kinouchi et al., BIOLOGICAL-ACTIVITIES OF THE INTESTINAL MICROFLORA IN MICE TREATED WITH ANTIBIOTICS OR UNTREATED AND THE EFFECTS OF THE MICROFLORA ON ABSORPTION AND METABOLIC-ACTIVATION OF ORALLY-ADMINISTERED GLUTATHIONE CONJUGATES OF K-REGION EPOXIDES OF 1-NITROPYRENE, Carcinogenesis, 14(5), 1993, pp. 869-874
To elucidate the effects of the intestinal microflora on absorption an
d activation of glutathione conjugates of 4,5-epoxy-4,5-dihydro-1-nitr
opyrene (1-NP 4,5-oxide) and 9,10-epoxy-9,10-dihydro-1-nitropyrene (1-
NP 9,10-oxide), we investigated the biological activities of the micro
flora in specific-pathogen-free (SPF) mice and SPF mice treated with v
arious antibiotics and established the methodology of antibiotic treat
ment to eliminate the intestinal microflora. Mice were given various k
inds of antibiotics by intragastric gavage twice a day for five days.
A mixture of antibiotics bacitracin (BC), neomycin (NM) and streptomyc
in (SM) was the most effective in reducing the various activities of t
he intestinal microflora. The treatment decreased the bacterial counts
and the activities of enzymes of the intestinal contents cysteine con
jugate beta-lyase (beta-lyase), beta-glucuronidase and nitroreductase
which were derived from the intestinal microflora, but did not affect
the activities of gamma-glutamyltransferase and aminopeptidase which w
ere derived from host tissue cells. Furthermore, the treatment did not
affect absorption of glucose from the intestinal tract, body weight o
r liver enzyme activities. The treatment with only an aminoglycoside a
ntibiotic, kanamycin or NM, decreased neither the number of anaerobes
in the intestine nor the beta-lyase or nitroreductase activities from
the intestinal contents. Glutathione conjugates of [H-3]-1-NP oxides w
ere administered to two groups of ICR mice that had been treated with
antibiotics (BC, NM, SM) or saline (control group) orally. The radioac
tivity in the blood increased and reached the maximum level 2 or 3 h a
fter administration of the conjugates in the control group; however, t
hat in the antibiotic-treated group was only slightly increased if at
all. Excretion of [H-3]-labeled metabolites into the urine was approxi
mately 20% of the total dose in the control group, but it was <2% in t
he antibiotic-treated group during 48 h. After 48 h, DNA in the lower
intestinal mucosa was extracted and the DNA adducts were analyzed by t
he P-32-postlabeling method. Three new DNA adducts were detected in th
e lower intestinal mucosa of the control group but not of the antibiot
ic-treated group. These results suggest that the intestinal microflora
plays an important role in absorption of the metabolites of glutathio
ne conjugates of 1-NP oxides from the intestinal tract and activation
of the metabolites in the intestine.