The role of a mixture of green tea, turmeric and chitosan in the treatment of obesity-related testicular disorders

J. Appl. Biomed.
5: 131–138, 2007
ISSN 1214-0287


ORIGINAL ARTICLE


The role of a mixture of green tea, turmeric and chitosan in the
treatment of obesity-related testicular disorders

Mohamed El-Sweedy1, Nabil Abdel-Hamid2, Mohamed El-Moselhy3

1Biochemistry Departments, College of Pharmacy, Zagazig University, Egypt
2College of Pharmacy, El-Minia University, Egypt
3Pharmacology Department, College of Pharmacy, El-Minia University, Egypt



Received 25th October 2006.
Revised 16th December 2006.
Published online 16th March 2007.



Summary
In the present investigation, we studied the effect of aqueous green tea extract (GTE), alcoholic
turmeric extract (ATE), and water-soluble Chitosan (WSC), individually/or in mixture, on the
testicular tissue content of total cholesterol (TC), triglycerides (TG), phospholipids (PL), and
thiobarbituric acid reactive substance (TBARS), in addition to nitric oxide (NO) in obese rats.
The testicular weight of the obese rats was increased more significantly than control; TC, TG, PL,
TBARS and NO were significantly higher in the obese group. GTE reduced testicular weight and
significantly reduced other estimated parameter. ATE significantly increased testicular weight, with
apparent peritesticular vascular congestion. It significantly decreased all other studied parameters.
WSC significantly increased testicular weight, with significant reduction of all other parameters. The
mixture of the three drugs non-significantly decreased testicular weight, and significantly decreased
other parameters, except NO, which was significantly more elevated than the obese control. We
concluded that obesity induced a significant increase in testicular weight, in addition to TC, TG, PL,
TBARS and NO, in comparison to the normal control subjects. An efficient protection against obesity-
induced changes was achieved by each individual drug, while the mixture of GTE, ATE and WSC
showed less protective potential than each individual drug.We here recommend the use of GTE, ATE
in treating obesity-related testicular dysfunction and suggest that attention should be paid to the
possible effect of WSC on the bioavailability of other concomitantly-used drugs and suggest a
pertinent clinical benefit of both GTE and ATE.


Keywords: obesity – green tea – turmeric – Chitosan – testes – rats




INTRODUCTION


Nabil Mohie Abdel-Hamid, Department of
Obesity is a common problem in affluent countries.
Biochemistry, Faculty of Pharmacy, El-Minia
Reduced energy expenditure from exercise or
University, El Minia, Egypt
metabolism or both, may be an important
nabilmohie@yahoo.com
contributory factor in developing obesity. Also,
+20506913997
failure in reducing food intake sufficiently to
+20106426998
maintain energy balance is a another strong cause.

Obese persons are at high risk of heart attack,
stroke, hypertension, diabetes mellitus, gall bladder

---

El-Sweedy et al.: Obesity-related testicular disorders



diseases and death (Bray and Gray 1988). More
(PL), thiobarbituric acid reactive substance
than 90% of body energy is stored as triglyceride in
(TBARS), in addition to nitric oxide (NO) in obese
adipose tissue. Protein and glycogen provide only
rats.
smaller quantities of energy. Triglycerides are

stored in fat cells that differ in size from one region

in the body to another (Bjorntorp and Ostman

1971).
MATERIALS AND METHODS
Rodents showed different genotypes for

obesity, some strains have autosomal dominant,
Animals and Experimental Design
others recessive or polygonal mutations (Festing
Sixty male Wistar rats, weighing 100–110 g, of 6–8
1979). Likewise, rare forms of human obesity have
weeks old, were obtained from Animal House,
a pure genetic basis (Bray 1989). In massively
Veterinary Medical College, University of Zagazig.
obese males, there is a decrease in testosterone, sex
The animals were fed a standard pellet diet
hormone binding globulin and free testosterone
(purchased from the same animal house) and
(Glass et al. 1981) and (Kley et al. 1979) It was
allowed free access to water for one week to
reported that spermatogenesis is disrupted by free
acclimatize. They were housed in a temperature-
radical toxicity. Thus, the study of oxidative stress
controlled room (temperature ranged from 19–
is a determinant in exploring some aspects
23 °C), with a 12 hours light and 12 hours dark
affecting fertility (Aruldhas et al. 2005).
cycle during the whole period of the experiment.
Changes in the testicular lipid profile, were
The standard feed used had the following
strongly correlated to testicular degeneration,
composition: 23.5% protein, 48.9% carbohydrate,
histochemical and biochemical disturbances
5% lipid, 12% water, 5.7% ash and 4% cellulose.
(Chowdhury et al. 1990). These changes were also
This diet continued only for the control group till
associated with increased lipid, DNA oxidative
the end of the experiment (Llado et al. 1995).
damage and depletion of lipid-soluble antioxidants
The animals were sub-divided into 6 equal
(Lucesoli and Fraga 1995).
groups, 10 animals/each. The remaining 5 groups
The use of herbal medicine increases every day
were fed high fat (cafeteria) diet for 35 days
and still finds a wide use worldwide. Traditional
(Geremias et al. 2006) This diet consisted of 9%
herbs have more acceptance than prescription drugs
protein, 21% carbohydrate, 13.6% lipid, 51.3%
in many cultures with emerging epidemics of
water and 5.1% fibers (Serra et al. 1987). The
obesity. This was mostly attributed to being safer
second group was fed only cafeteria diet, served as
than drugs, from the point of view of patients. Also
obese control and was not given drugs. The third
patients believe that by using this type of
group was given green tea (Camellia sinensis), 2%
medication there is no need for a physician, and it
aqueous extract as drinking liquid, simultaneously
may be a relevant attempt to compensate for drug
with the cafeteria diet (Challa et al. 1997). The
failure in managing obesity. Some of these
fourth group was given alcoholic turmeric extract
alternatives were, green tea and a binding resin
(Curcuma longa) as (25 mg/kg), daily by
(chitosan), which can precipitate lipids, leading to
intragastric tube. This extract was prepared by
depressed fat absorptivity (Heber 2003). Different
extracting 500 g of chopped rhizomes with 1.5
studies showed that both green and black tea
liters of 95% ethanol, by soaking overnight,
contain flavonoids such as quercetin, kaempferol
filtering, re-extracting the sediment and filtering
and myricetin, which are potent anti-carcinogenics
again. Both filtrates were mixed, evaporated in a
(Herlog et al. 1993). It also reduced the risk of
rotavapor at 40–50 ?C, under reduced pressure, re-
colon, stomach, lung and skin cancers (Challa et al.
dissolved into alcohol to give a final concentration
1997).
of 40 mg/dl, and then the dose was calculated to 25
The use of turmeric extract in a group of herbs
mg/kg, body weight (Saravanan and Pari 2005).
as a treatment for diabetes, induced fatty liver and
The fifth group was given WSC as 4% in the
showed a significant reduction in triglyceride, total
cafeteria diet (Geremias et al. 2006).
cholesterol and phospholipids-content in diabetic
The sixth group was given a mixture of the
rats (Saravanan and Pari 2005).
three drugs in the same mentioned doses. All drugs
The present study was mainly conducted to
were given by intragastric tube as a single daily
study possible metabolic disturbances in rat
dose for 35 days.
testicular tissue after induction of obesity, and
After 35 days, on the morning of the next day,
whether some known anti-obesity natural products,
the animals were killed, the testicles were removed,
as GTE, ATE and a well-defined carbohydrate
kept frozen in liquid nitrogen at –80 ?C to the day
polymer (WSC) – if used individually or together –
of analysis.
may have a corrective potential in relation to

testicular dysfunction. This was achieved by an
Drugs and chemicals
estimation of the testicular content of total
All drugs and chemicals were of analytical grade
cholesterol (TC), triglycerides (TG), phospholipids
and purchased from local suppliers.

132

---

El-Sweedy et al.: Obesity-related testicular disorders



Methods
RESULTS
The testes were thawed, weighed (as wet weight),

the gross features were observed, recorded,
Table 1 shows that feeding the animals with
decapsulated and cut into small pieces (Wang et al.
cafeteria diet increased testicular weight
1997). Lipids were extracted from tissues by the
significantly more than the control. It also
method of Folch et al. (1957). TC was estimated by
significantly elevated the testicular content of TC,
the method of Zlatkis et al. (1953). To 0.1 ml of the
TG, PL, TBARS and NO. Treatment with GTE
lipid extract, 9.9 ml of ferric chloride – acetic acid
significantly decreased these values, in comparison
reagent were added, mixed, allowed to stand for 15
to obese rats, except TC, which was left to increase
minutes and centrifuged. 5 ml of the supernatant
significantly more than than the obese control. ATE
were mixed with 3 ml conc. sulfuric acid. The
significantly induced peri-testicular congestion in
developed color was read after 20 minutes at 560
the veins of all treated animals. It significantly
nm against the reagent blank.
increased testicular weight compared to the obese
TG were determined by the method of Bucolo
control; and it significantly decreased other
and David (1973) in an aqueous liquid prepared
parameters in comparison to obese control.
from the lipid extract (chloroform/methanol, 2:1).
Chitosan treatment significantly increased testicular
A certain volume of the lipid extract was
weight compared to obese control), but also
evaporated in a boiling water bath, the sediment
significantly decreased other values compared to
was dissolved in 0.2% Triton X100 to give an
obese control subjects. The mixture of the three
aqueous medium (Ide et al. 2004).
drugs non-significantly elevated testicular weight
The phospholipid content was colorimetrically
compared to obese control, but significantly
determined in the total lipid extract, depending on
decreased TC, TG, PL, TBARS, although it
the phosphorus content, without acid digestion, to
significantly elevated NO content compared to
exclude interference of inorganic phosphorus
obese control.
(Raheja et al. 1973). Egg yolk was extracted by

chloroform/methanol, and serially diluted for

preparation of the standard curve, from which the
DISCUSSION
PL content was calculated.

Nitric oxide was extracted from the tissue in
The base of information about obesity has
100 mM phosphate buffer, pH 7.4, containing 17
dramatically expanded in the last three decades.
mM sulfanilamide and 0.4 mM N–(1–naphthyl)
The importance of fat distribution as a health risk
ethylenediamine dihydrochloride as described
has added a new dimension to this problem (Bray
elsewhere (Nims et al. 1995). NO was then
and Gray 1988). It was proved that hormone-
spectrophotometrically-determined utilizing copper
sensitive lipase (HSL) is essential for
– cadmium alloy as a reducing agent. This method
spermatogenesis. This enzyme is necessary for
basically relied on the reduction of nitrate into
hydrolysis of triacylglycerol and cholesteryl esters
stable nitrite (Gries reaction) (Sastry et al. 2002). A
in many tissues, including ovaries and testes. This
standard curve was obtained from different
role of the enzyme shed light on the importance of
concentrations of sodium nitrite (Bauche et al.
lipids in studying testicular derangements (Osuga et
1998).
al. 2000). Nevertheless, the absence of HSL from
Lipid peroxidation product (malondialdehyde)
Leydig cells (testicular cells, responsible for
was colorimetrically estimated as TBARS
testosterone production) did not affect the plasma
according to the method of Niehius and
testosterone level. Thus, oligospermia did not result
Samuelsson (1968). Briefly, 0.1 ml of tissue
from hypogonadism in some isolated testicular
homogenate in Tris-HCl buffer (pH 7.5) was
cells in vitro (Sassone–Corsi 1979).
treated with 2 ml of thiobarbituric acid 0.37–15%
This controversial testosterone level depending
trichloroacetic acid and 0.25 N HCl, 1:1:1 (TBA –
/or non-dependent on HSL was a reason why serum
TCA – HCl) reagent. The mixture was then placed
testosterone level was not estimated in the present
in water bath for 15 minutes, cooled and the
work. In the present study, the induction of obesity
absorbance of the clear supernatant was measured
lead to a significant increase in testicular weight,
at 535 nm.
TC, TG, PL, TBARS and NO contents. This

variation was also recorded before in some strains

of mice, compared to their homozygous lean
Statistical analysis
controls, in the form of lipid accumulation,
The data were analyzed using analysis of variance
associated with decreased lipolysis in isolated
(ANOVA) and the group means were compared by
Leydig cells. These changes in the testicular
Duncan's Multiple Range Test (DMRT). Values of
endocrine function of obese mice were interpreted
p<0.05 were considered statistically significant
as a possible consequence of pituitary dysfunction
(Duncan 1957).
(Khun-Velten et al. 1986).


133

---

El-Sweedy et al.: The role of a mixture of green tea

Table 1: Variations in testicular total cholesterol, triglycerides, phospholipids, thiobarbituric acid reactive substance
and nitric oxide in obese rats, treated with green tea, turmeric extracts and chitosan for 35 day (values are given as
mean±SEM, n=10)


Group
Control Obese Green tea Turmeric
Chitosan Green
tea
control
+ Turmeric
Parameter
+ Chitosan







Weight of single
0.87
0.99*
0.93
1.30#
1.10#
0.96
testis (g )
±0.03
±0.05
±0.02
±0.02
±0.03
±0.01







Total Cholesterol
11.3
14.7*
17.1#
7.2#
9.8#
12.2#
(mg/g wet tissue)
±1.1
±0.1
±0.1
±0.1
±0.1
±0.1







Triglycerides
31.10
40.00*
20.00#
11.10#
9.30
4.00#
(mg/g wet tissue)
±0.73
±0.80
±2.40
±0.60
±0.32#
±0.32







Phospholipids
43.20
100.00*
25.40#
35.10#
17.60#
56.80#
(mg/wet tissue)
±0.53
±1.10
±0.37
±0.48
±0.33
±0.70







TBARS
403.0
546.6*
358.1#
366.0#
433.0#
388.1#
(nmol/g wet tissue)
±2.1
±3.4
±3.4
±4.1
±4.2
±4.1







Nitric Oxide
490.0
625.1*
180.0#
536.1#
586.0#
802.0#
(nmol/g wet tissue)
±3.6
±3.8
±2.5
±3.1
±3.8
± 2.8


* Significantly different from control
# Significantly different from obese control





The increased cholesterol levels were reported to
on steroidogenesis by Leydig cells, both in vivo
be an important risk of testicular cancer (Dobson
(Adams et al. 1994) and in vitro (Del Punta et al.
2005)
1996). Thus, decreasing NO level may activate
Spermatozoa are rich in polyunsaturated fatty
spermatogenesis, that may be inhibited by obesity-
acids and more liable for lipid peroxidation by
related NO accumulation.
reactive oxygen species (ROS).
Our results revealed that turmeric treatment
The oxidation product (TBARS) increases in
significantly decreased TC, TG, PL, TBARS and
most spermatogenic disturbances (Sharma and
NO contents, although it significantly increased
Agarwal 1996). Increased testicular lipids, in
testicular weight, in comparison to obese rats. The
conjunction with obesity may lead – in addition to
decreased content of lipids concomitantly with
varices – to obstructive azospermia, which was
elevated weight, is most probably due to the
reported to be associated with increased tissue
prominent congestion of peri-testicular veins. This
nitrite and TBARS (Basar et al. 2006).
congestion wasn't clear in other groups. Turmeric
Treatment with GTE significantly decreased
has many active components, but curcumin is the
testicular TG, PL, TBARS, NO and non-
most potent ingredient. It is a powerful ani-
significantly testicular weight, but couldn't retain
inflammatory and anti-oxidant and has greater
TC content. It was shown that testicular cholesterol
effects in preventing free radical damage, compared
exists in three different forms: free, ester and
with vitamins C, E and superoxide dismutase
sulfate. The free form is about 91% at all ages,
(Sharma 1976).
which increases by maturation (Connor et al.
This anti-oxidant activity is clear on the
1997). The antioxidant activity of GTE was
testicular level, which is manifested in decreased
previously reported in mice. It also protected DNA
TBARS and NO contents. This will correct the
from oxidative damage (Shi et al. 1994).
possible inhibitory effect of elevated NO on Leydig
It also inhibited both lung (Wang et al. 1992)
cell steroidogenesis, that may be inhibited by
and liver (Wang et al. 1988) carcinomas. Increased
obesity (Adams et al. 1994). The elevated testicular
NO generation in testes showed an inhibitory effect
TC content induced by obesity may be due to
134

---

El-Sweedy et al.: Obesity-related testicular disorders



impaired utilization in steroidogenesis, which may
content, if compared to obese rat values. As
have been corrected by turmeric administration
previously stated, the effect on testicular weight, in
(Lin et al. 1995). The changes in lipid content seem
spite of decreasing lipid figures is attributed to
to be more apparent in PL. PL are always more
another pathway for chitosan, by which tissue
prevalent than TC and TG in testicular tissue
protein may be increased and consequently also
(Oshima and Carpenter, 1968).
tissue weight (Poropratto et al. 2005).
The role of turmeric in testicular protection
The idea of polyherbal formulation in medical
may also be referred to its anti-apoptotic property
practice was extensively applied a long time ago,
(Mohanty et al. 2006). In the present work,
including GTE for management of obesity (Saper
chitosan treatment significantly decreased testicular
et al. 2004). Mostly, the presence of WSC in the
TC, TG, PL, TBARS and NO contents, but
mixture used in this study didn't elicit effects as
elevated testicular weight significantly more than
satisfactory as if each individual component had
obese subjects. From the results shown in the
been used solely. Although WSC alone was
Table, it seems that increased testicular weight is
reported to adjust the metabolic functions
not correlated to fat content which was
controlling fertility (Choi et al. 2002), it seems that
significantly depressed.
addition of other herbs may decrease this potential.
It was reported that oral administration of
On the other hand, WSC alone increased NO
WSC to rats whether alone or mixed with aloe vera
production, but it was more synergistically
extract could prevent the atherogenic process
increased when an additional drug was
associated with hyperlipidaemia by depressing
simultaneously used as interferon – gamma (IFN –
blood levels of TC, TG, low density lipoprotein and
gamma) in an in vitro study (Seo et al. 2000). The
very low density lipoprotein cholesterol (Geremias
previous results on chitosan-containing mixtures
et al. 2006). It is important to point out that WSC is
concerning NO production are in agreement with
not an efficient drug for treating obesity, but it is a
our results
preventive medication, that can inhibit fat
The presence of ATE in the mixture, mostly
absorption (Choi et al. 2002) and without dietary
augmented the effect of WSC, due to the
surveillance, it will not be efficient (Ho et al.
antioxidant, adaptogenic, ani-inflammatory and
2001).
anti-infectious activities of its curcumin content
Chitosan is a natural non-toxic polysaccharide,
(Srinivas 1992). These effects, taken together,
havinga chemical composition as poly-N-acetyl
improve fertility and testicular performance,
glucosaminoglycan, which is a bioabsorbable
through controlling both lipoperoxidation and NO
polymer known to accelerate wound healing
production, which simultaneously affect sperm
(Ozmeric et al. 2000). In our study, chitosan
motility (Romeo et al. 2003).
increased testicular weight, whether taken alone or

in mixture. It decreased lipid content, so the

increased weight may be attributed to increased

protein content at the expense of lipid value. This is
CONCLUSION
most probably because WSC has been reported to

interact with cell membranes, enhancing peptide
We thought that obesity can be considered as an
and protein uptake, but interfering with lipid uptake
important contributory factor underlying testicular
(Poropratto et al. 2005). On the other hand, WSC
dysfunction. This is clearly shown by increased
showed an activation of intestinal immune
testicular weigh, concomitantly with increased TC,
functions and prevented tumor growth, probably
TG, PL, elevated oxidative stress indicated by
through activation of natural killers and chemotaxis
increased lipoperoxidation and NO production.
(Maeda and Kimura 2004). This effect, in addition
Each drug when used individually, corrected these
to antioxidant properties – seen in our work – can
testicular parameters in the direction of improving
be considered as a synergistic benefit of its
testicular function. The most protective drug that
networking pathways.
alleviated oxidative stress, was GTE, if compared
In another study, WSC showed potent
with ATE and WSC. All three drugs individually
antioxidant properties in tissues by decreasing
decreased testicular lipid profile in obese animals.
TBARS and increasing antioxidant enzymes,
The use of a mixture containing chitosan mostly
catalase and superoxide dismutase (Jeon et al.
did not introduce any more observable benefit than
2003). This protective action against some
individual drugs, mostly due to the incompatibility
hepatotoxic chemicals was also noticed by
of both extracts to chitosan polymers. Although our
inhibiting malondialdehyde formation triggered by
results were designed on laboratory animals,
carbon tetrachloride (Yan et al. 2006). In the
human studies may introduce more satisfactory and
present study, we noticed that the use of GTE, ATE
reliable results. We can recommend the use of GTE
with WSC induced a non-significant decrease in
and ATE for treating most cases of expected
testicular weight, a significant decrease in TC, TG,
testicular dysfunction, whether in obese, or non-
PL, TBARS and a significant increase in NO
obese individuals. However, care should be

135

---

El-Sweedy et al.: Obesity-related testicular disorders



exercised in prescribing chitosan with drugs that
Desmosterol and docosahexanoic acid. J. Clin.
may be affected by WSC polymer form, that might
Endocrinol. Metab. 82:1911–1916, 1997.
lead to poor bioavailability and usefulness.
Del Punta K, Charreau EH, Pignataro OP: Nitric

oxide inhibits Leydig cell steroidogenesis.

Endocrinology 137:5337–5343, 1996.

Dobson R: High cholesterol may increase the risk

of testicular cancer. BMJ 330:1042, 2005.
REFERENCES
Duncan BD: Multiple range test for correlated

heteroscedastic means. Biometrics 13:359–
Adams ML, Meyer ER, Sewing BN, Ciuro TJ:
364, 1957.
Effects of nitric oxide-related agents on rat
Festing MFW (ed): Animal Models of Obesity.
testicular dysfunction. J. Pharmacol. Exp.Ther.
Oxford Univ Press, NY 1979.
269:230–237, 1994.
Folch J, Lees M, Sloane-Stanley GH: A simple
Aruldhas M, Subramanian S, Sekar P,
method for the isolation and purification of
Vengatesh G, Chandrahasan G, Govindarajulu
total lipids from animal tissues. J. Biol. Chem.
P, Akbarsha MA: Chronic chromium
226:497–509, 1957.
exposure-induced changes in testicular
Geremias R, Pedrosa RC, Locatelli C, de Favere
histoarchitecture are associated with oxidative
VT, Coury-pedrosa R, Laranjeira MC: Lipid
stress: study in a non-human primate (Macaca
lowering activity of hydrosoluble chitosan and
radiata Geoffroy). Hum. Reprod. 20:2801–
association with Aloe vera L and Brassica
2813, 2005.
olearacea L. Phytother. Res. 20:288–293,
Basar MM, Kisa Ü, Tuglu D, Yilmaz E, Basar H,
2006.
Caglayan O, Batislam E: Testicular nitric
Glass AR, Burman KD, Dahms WT, Boehm TM.:
oxide and thiobarbituric acid reactive oxygen
Progress in endocrinology and metabolism:
substance levels in obstructive azoospermia: A
Endocrine function in human obesity.
possible role in pathophysiology of infertility.
Metabolism 30:89–104, 1981.
Mediators of inflamm. article ID: 27458, pp.
Heber D: Herbal preparations for obesity: are they
1–5, 2006.
useful? Prim. Care 30:441–463, 2003.
Bauche F, Stephan JP, Touzalin AM, Jegou B: In
Herlog MGL, Hollman PCH, Putte BV: Content of
vitro regulation of an inducible type NO
potentially carcinogenic flavonoids of tea
synthase in the rat seminiferous tubule cells.
infusion, wines and fruit juices. J. Agric. Food
Biol. Reprod. 58:431–438, 1998.
Chem. 41:1242–1246, 1993.
Björntorp P, Ostman J: Human adipose tissue
Ho HC, Tai ES, Eng PH, Tan CE, Fork AC: In the
dynamics and regulation. Adv. Metab. Disord.
absence of dietary surveillance, chitosan does
5:277–327, 1971.
not reduce plasma lipids or obesity in
Bucolo G, David H: Quantitative determination of
hypercholesterolaemic obese Asian subjects.
serum triglycerides by the use of enzymes.
Singapore Med. J. 42:6–10, 2001.
Clin. Chem. 19:476–482, 1973.
Ide T, Tsunoda M, Mochizuki T, Murakami K:
Bray GA: Classification andevaluation of obesities.
Enhancement of insulin signaling through
Med. Clin. North Am. 73:161–184, 1989.
inhibition of tissue lipid accumulation by
Bray GA, Gray DS: Obesity: Part I: Pathogenesis.
activation of peroxisome proliferator-activated
West. J. Med. 149:429–441, 1988.
receptor (PPAR) in obese mice. Med. Sci.
Challa A, Rao DR, Reddy BS: Interactive
Monit. 10:BR388–BR395, 2004.
suppression of abberant crypt foci induced by
Jeon TI, Hwang SG, Park NG, Jung YR, Shin SI,
azoxymethane in rat colon by phytic acid and
Choi SD, Park DK: Antioxidative effect of
green tea. Carcinogenesis 18:2023–2026,
chitosan on chronic carbon tetrachloride
1997.
induced hepatic injury in rats. Toxicology
Choi HG, Kim JK, Kwak HD, Cho JR, Kim TY,
187:67–73, 2003.
Kim BJ, Jung KY, Choi BK, Shin MK, Choo
Kley HK, Solbach HG, McKinnan JC,
YK: Effects of high molecular weight water-
Kruskemper
H: Testosterone decrease and
soluble chitosan on in vitro fertilization and
estrogen increase in male patients with obesity.
ovulation in mice fed a high-fat diet. Arch.
Acta Endocrinol. 91:553–563, 1979.
Pharmacol. Res. 25:178–183, 2002.
Kuhn-Velten N, Codjambopoulo P, Haider SG,
Chowdhury AR, Gautam AK, Bhatnagar VK:
Passia D, Kley HK, Herberg L, Staib W,
Lindane-induced changes in morphology and
Goslar HG: Biochemical and histochemical
lipid profile of testes in rats. Biomed. Biochim.
studies on the pituitary-testicular axis in obese
Acta 49:1059–1065, 1990.
(C57Bl/6J.ob/ob) mice. Int. J. Androl. 9:123–
Connor WE, Lin DS, Neuringer M: Biochemical
131, 1986.
markers for puberty in the monkey testes:

136

---

El-Sweedy et al.: Obesity-related testicular disorders



Lin DS, Connor WE, Wolf DP, Alexander M:
oxidative damage in adolescent variocele.
Uneven distribution of monkey sperm.
Hum. Reprod. 18:26–29, 2003.
FASEB J. 9:A 81, 1995.
Saper RB, Eisenberg DM, Phillip RS: Common
Llado I, Pico C, Palou A, Pons A: Protein and
dietary supplements for weight loss. Am. Fam.
amino acid intake in cafeteria fed obese rats.
Physician 70:1731–1738, 2004.
Physiol. Behav. 58:513–519, 1995.
Saravanan R, Pari L: Antihyperlipidemic and
Lucesoli F, Fraga CG: Oxidative damage to lipids
antiperoxidative effect of Diasulin, a
and DNA concurrent with decrease of
polyherbal formulation in alloxan induced
antioxidants in rat testes after acute iron
hyperglycemic rats. BMC Complement.
intoxication. Arch. Biochem. Biophys.
Altern. Med. 5:14–21, 2005.
316:567–571, 1995.
Sassone-Corsi P: Transcriptional checkpoints
Maeda Y, Kimura Y: Antitumor effects of various
determining the fate of male germ cells. Cell
low-molecular weight chitosans are due to
88:163–166, 1997.
increased natural killer activity of intestinal
Sastry KVH, Moudgal RP, Mohan J, Tyagi JS,
intra-epithelial lymphocytes in sarcina-180-
Rao GS: Spectroph-otometric determination of
bearing mice. Nutrition 134:945–950 , 2004.
serum nitrite and nitrate by copper-cadmium
Mohanty I, Arya S, Gupta SK: Effect of Curcuma
alloy. Anal Biochem. 306:79–82, 2002.
longa and Ocimum sanctum on myocardial
Seo WG, Pae HO, Kim NY, OH GS, Pak IS, Kim
apoptosis in experimentally-induced
YH, Kim YM, Lee YH, Jun CD, Chung HT:
myocardial ischemic – reperfusion injury.
Synergistic cooperation between water-soluble
BMC Complement. Altern. Med. 6:3–14,
chitosan oligomers and interferon-gamma for
2006.
induction of nitric oxide synthesis and
Niehius WG, Samuelsson D: Formation of
tumoricidal activity in murine peritoneal
malondialdehyde from phospholipids
macrophages. Cancer Lett., 159:189–195,
arachidonate duringmicrosomal lipid
2000.
peroxidation. Eur. J. Biochem. 6:126–130,
Serra F, Bonet T, Palou A: Amino acid enzyme
1968.
activities in brown and white adipose tissues
Nims RW, Darbyshire JF, Saavedra JE,
and the liver of cafeteria rats: effects of 24
Christodoulou D, Hanbauer I, Cox GW,
hours starving. Arch. Int. Physiol. Biochim.
Grisham MB, Laval F, Cook, JA, Krishna,
95:263–268, 1987.
MC, Wink D.A: Colorimetric methods for the
Sharma OP: Antioxidant properties of curcumin
determination of nitric oxide concentration in
and related compounds. Biochem. Pharmacol.,
neutral aqueous solutions. Methods: A
25:1811–1825, 1976.
Companion Methods in Enzymology 7:48–54,
Sharma RK, Agarwal A: Role of reactive oxygen
1995.
species in male infertility. Urology 48:835–
Oshima M, Carpenter MP: The lipid composition
850, 1996.
of the prepubertal and adult rat testis .
Shi TS, Wang ZY, Smith TJ, Hong JY, Ho CT,
Biochim. Biophys. Acta 152:479–497, 1968.
Yang CS: Effects of green tea and black tea on
Osuga J, Ishibashi S, Oka T, Yagyu H, Tozawa R
4-(methylnitrosamino)-1-(3-pyridyl)-1-
et al.: Targeted disruption of hormone-
butanone bioactivation, DNA methylation and
sensitive lipase results in male sterility and
lung tumorigenesis in A/J mice. Cancer Res.
lipocyte hypertrophy, but not in obesity. Proc.
54:4641–4647, 1994.
Natl. Acad. Sci. U.S.A. 97:787–792, 2000.
Srinivas L, Shalini VK, Shylaja M: Turmerin: A
Ozmeric N, Ozcan G, Haytac CM, Alaadinoglu EE,
water-soluble antioxidant peptide from
Sargon MF, Senel S: Chitosan film enriched
turmeric (Curcuma longa). Arch. Biochem.
with an antioxidant agent, taurine, in
Biophys. 292:617–623, 1992.
fenestration defects. J. Biomed. Mater. Res.
Wang ZY, Bickers DR, Mukhtar H: Interction of
51:500–503, 2000.
epicatechin derived from green tea with rat
Poropratto C, Bianco ID, Correa SG: Local and
hepatic cytochrome P-450. Drug Metabol.
systemic activity of the polysaccharide
Disposal 16:93–103, 1988.
chitosan at lymphoid tissues after oral
Wang ZY, Hong MT, Reuhl KR, Conney AH,
administration. J. Leukoc. Biol. 78:62–69,
Yang CS: Inhibition of N-nitrosodi-
2005.
methylamine and 4-(methylnitrosamine)-1-(3-
Raheja RK, Charanjik K, Singh A, Bhatia JS: New
pyridyl)-1-butanone-induced tumorigenesis in
colorimetric method for estimation of
A/J mice by green tea and black tea. Cancer
phospholipids without acid digestion. J. Lipid
Res. 52:1943–1947, 1992.
Res. 14:695–697, 1973.
Wang SW, Chiao YC, Tsai SC et al.: Inhibition of
Romeo C, Ientile R, Impellizzeri P et al.:
bufalin on pituitary and testicular function in
Preliminary report on nitric oxide-mediated
rats. J. Pharmacol. Exp. Ther. 283:528–532,
1997.

137

---

El-Sweedy et al.: Obesity-related testicular disorders



Yan Y, Wanshun L, Baoqin H, Bing L, Chenwei F:
Zlatkis A, Zak B, Boyle G: A method for the
Protective effects of chitosan
determination of serum cholesterol. J. Clin.
ligopolysaccharide and its derivatives against
Med. 41:486–492, 1953.
carbon tetrachloride-induced hepatic damage
in rats. Hepatol. Res. 35:178–184, 2006.


138

---