122 Fluoride 2005;38(2):122–126 Research report
RECOVERY FROM FLUORIDE+ALUMINIUM TOXICITY IN VAS
DEFERENS, SEMINAL VESICLE, AND VENTRAL PROSTATE
OF MICE BY VITAMIN C
NJ Chinoy,a R Momin, HP Sorathia, DD Jhala
Ahmedabad, India
SUMMARY: Combined administration of sodium fluoride (NaF, 10 mg/kg bw/
day)+aluminium chloride (AlCl3, 200 mg/kg bw/day) to adult male mice for 30 days
caused histological changes in the vas deferens. A decrease in protein in the vas
deferens, seminal vesicles, and prostate occurred. Inhibition of phosphorylase in
vas deferens together with accumulation of glycogen altered its carbohydrate
metabolism. Seminal vesicle fructose and ventral prostate acid phosphatase
(ACPase) also decreased causing changes in their secretions necessary for sperm
function. Withdrawal of treatment for 30 days led to partial recovery of these
parameters, whereas treatment with NaF+AlCl3+vitamin C brought about complete
recovery.
Keywords: Acid phosphatase; Aluminium and accessory sex glands; Fluoride and accessory
sex glands; Fructose; Glycogen metabolism; Mice ventral prostate; Seminal vesicle; Vas
deferens.
INTRODUCTION
The effects of aluminium+fluoride toxicity in testis and epididymis of mice
after 30 days have been reported earlier.1-6 Withdrawal of treatment caused some
recovery, but treatment with vitamin C together with the toxicants facilitated
recovery in the respective organs. The effects of these toxicants on the vas deferens, which has absorptive, synthesizing, and secretory ability for maintenance of
sperm in a viable state,7 as well as their effects on the seminal vesicles and prostate have not been adequately investigated. Hence, the present study was undertaken.
MATERIALS AND METHODS
Healthy, adult, pathogen–free, colony bred male albino mice (Mus musculus) of
Swiss strain were procured and treated as described earlier5 and outlined in the
following protocol:
Group
Treatment and dose
(10–12 animals in each group)
Duration
(days)
Day of
autopsy
I
Control+distilled water (DW)
-
a
II
NaF–treated (10 mg/kg bw)+AlCl3–treated (200 mg/kg bw)
30
31st
III
Same as in Group II then withdrawal for an additional 30 days
30+30
61st
IV
Same as in Group II+Vitamin C (15 mg/animal/day) for 30 days
30+30
61st
a
Sacrificed along with treated mice.
aFor
correspondence: Reproductive Endocrinology and Toxicology Unit, Department of
Zoology, School of Sciences, Gujarat University, Ahmedabad-380 009, India.
E-mail: [email protected]
Fluoride 2005;38(2)
Recovery from fluoride+aluminium toxicity in accessory sex glands of mice 123
After the respective treatments, the animals were sacrificed by cervical dislocation. The vas deferens, seminal vesicles, and ventral prostate were excised, blotted free of blood, and utilized for the study. Histology of vas deferens was carried
out by haematoxyline and eosin (HE) staining. The levels of protein8 in all three
organs, and glycogen9 and phosphorylase10 in vas deferens, fructose11 in the
seminal vesicles, and acid phosphatase12 in the ventral prostate were determined
using the methods cited.
Statistical analysis: For all biochemical parameters a minimum of 5 or 6 replicates were assayed, and the data were statistically analysed by Student’s t test.
RESULTS
Vas deferens histology: The histology of control mice vas deferens showed the
epithelial folds with pseudostratified epithelium having stereocilia. Muscle layers
were also seen (Figures 1 and 2). The treatment with NaF+AlCl3 for 30 days in
mice caused epithelial cell pycnosis, clumping of stereocilia, and absence of
sperm bundles in the lumen (Figures 3 and 4). Withdrawal of NaF+AlCl3 treatment for 30 days (Group III) showed some recovery with the presence of stereocilia and sperm bundles (Figures 5 and 6). Administration of ascorbic acid along
with NaF and AlCl3 (Group IV) for 30 days revealed no toxic effects since the
structure of the vas deferens then appeared normal (Figures 7 and 8).
Biochemical parameters: The levels of protein decreased significantly
(p<0.001) in all organs studied. Similarly, the levels of fructose in the seminal
vesicles, the activities of acid phosphatase in the ventral prostate and phosphorylase in the vas deferens also declined significantly (p<0.001) in Group II animals
as compared to Group I controls (Table). However, glycogen levels were
increased in the vas deferens of Group II mice as compared to Group I.
The withdrawal of treatment caused recovery in all parameters with significance levels ranging from p<0.05 to p<0.01 (Table). On the other hand, vitamin C
treatment along with NaF+AlCl3 in Group IV mice promoted a very significant
recovery (p<0.001) in all organs.
Figure 1. Transverse section of vas deferens
of control Group I mouse.
HE staining (X 200).
Figure 2. Magnified view of Figure 1.
HE staining (X 800).
Fluoride 2005;38(2)
124 Chinoy, Momin, Sorathia, Jhala
Figure 3. Transverse section of vas deferens
of Group II (NaF+AlCl3) treated mouse.
HE staining (X 200).
Figure 4. Magnified view of Figure 3.
HE staining (X 800).
Figure 5. Transverse section of vas deferens
of Group III mouse showing some recovery.
HE staining (X 225).
Figure 6. Magnified view of Figure 5.
HE staining (X 850).
Figure 7. Transverse section of vas deferens
of Group IV(NaF+AlCl3+vitamin C) treated
mouse showing marked recovery.
HE staining (X 200).
Figure 8. Magnified view of Figure 7.
HE staining (X 800).
Fluoride 2005;38(2)
Recovery from fluoride+aluminium toxicity in accessory sex glands of mice 125
Table. Protein (mg/100 mg fresh tissue wt) in vas deferens, seminal vesicle and ventral
prostate, vas deferens glycogen (µg/mg fresh tissue wt) and activity of phosphorylase (µg
phosphorus released/mg protein/15 minutes), seminal vesicle fructose (µg/mg fresh tissue wt) and acid phosphatase (ACPase) (µmoles of p-nitro phenyl liberated/mg protein)
in ventral prostate of Groups I to IV micea
Parameter
Organ
Group I
Group II
0.01§
Group III
6.33 ± 0.14§
Protein
Vas
deferens
9.63 ± 0.21
4.37 ±
Protein
Seminal
vesicle
6.13 ± 0.03
3.30 ± 0.80§
5.09 ± 0.07‡
6.07 ± 0.04§
Protein
Ventral
prostate
9.45 ± 0.11
5.74 ± 0.42§
7.08 ± 0.46*
8.50 ± 0.25§
Glycogen
4.97 ±
Group IV
0.27*
Vas
974.16 ± 13.9 1468.34 ± 45.3§ 1326.92 ± 41.49† 1032.83 ± 16.08§
deferens
PhosVas
phorylase deferens
8.09 ± 0.01
5.18 ± 0.15§
5.98 ± 0.33*
7.04 ± 0.09§
Fructose
Seminal
vesicle
13.61 ± 0.08
9.87 ± 0.76§
11.99 ± 0.19†
12.21 ± 0.07§
ACPase
Ventral
prostate
0.22 ± 0.007
0.13 ± 0.01§
0.19 ± 0.03*
0.21 ± 0.02§
a
Data are expressed as mean ± SE; * P<0.05; † P<0.02; ‡ P<0.01; § P<0.001; where no sign = not
significant
Comparisons: Group I with Group II; Group II with Group III; Group II with Group IV individually.
DISCUSSION
At the dose levels used in the study the protein levels in all organs (vas deferens, seminal vesicle and ventral prostate) were significantly decreased, which
might account for the decline in their enzyme activities and observed structural
alterations, especially in the vas deferens such as epithelial cell pycnosis and
clumping of stereocilia. Similar data were reported earlier using a different protocol and dose regimens of NaF and/or AlCl3 in mice.3,4
The accumulation of glycogen along with inhibition of phosphorylase evidently
reflects a disturbance of carbohydrate metabolism in vas deferens leading to
abnormal sperm metabolism in agreement with earlier work.3,4 In Group II the
decrease in acid phosphatase, a marker enzyme for prostate function, suggests
changes in prostate metabolism,13 and the decrease in fructose levels in the seminal vesicles of mice indicates alteration in testosterone levels in agreement with
findings of others.2-4
As reported in earlier communications,3-6 the withdrawal of treatment helped in
partial recovery in all parameters, whereas addition of vitamin C resulted in complete recovery in all organs, probably because of its antioxidant, reducing properties leading to an increase in C-AMP, which promotes growth and metabolism.3,4
Fluoride 2005;38(2)
126 Chinoy, Momin, Sorathia, Jhala
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