Content uploaded by Rohitash Jamwal
Author content
All content in this area was uploaded by Rohitash Jamwal on Aug 30, 2018
Content may be subject to copyright.
Effect of Cinnamomum cassia Methanol Extract and
Sildenafil on Arginase and Sexual Function of Young Male
Wistar Rats
Sumanta K. Goswami, PhD,* Mohammed N. Inamdar, PhD,* Rohitash Jamwal, M.Tech,†and
Shekhar Dethe, PhD†
*Department of Pharmacology, Al-Ameen College of Pharmacy, Bangalore, India; †Bioassay Lab, Research and
Development Centre, Natural Remedies Private Limited, Bangalore, India
DOI: 10.1111/jsm.12535
ABSTRACT
Introduction. Herbs have been used as an aphrodisiac since ages. Cinnamomum cassia is an important ingredient of
many Ayurvedic formulations to treat male sexual disorder including erectile dysfunction (ED).
Aim. The objective of the present study was to evaluate erectogenic and aphrodisiac activity of methanol extract of
C. cassia bark in young male rats.
Methods. Methanol extract of C. cassia was screened in vitro for arginase inhibition potential and IC50 was deter-
mined. Effect of the extract was observed in vitro on phenylephrine pre-contracted isolated rat corpus cavernosum
smooth muscle (CCSM) at 0.1, 1, 10, and 100 μg/mL. Young male Wistar rats were dosed with extract at 100 mg/kg
body weight for 28 days and its effects on sexual behavior and penile smooth muscle : collagen level were observed.
Main Outcome Measure. Effect of C. cassia was studied on arginase activity in vitro and sexual behavior of young
male rats.
Results. C. cassia inhibited arginase activity in vitro with an IC50 of 61.72 ±2.20 μg/mL. The extract relaxed
phenylephrine pre-contracted isolated rat CCSM up to 43% and significantly increased (P<0.05) sexual function of
young male rats. Treatment with the extract also increased smooth muscle level and decreased collagen level in rat
penile tissue.
Conclusion. The study proves usefulness of methanol extract of C. cassia bark for increasing sexual function.
Goswami SK, Inamdar MN, Jamwal R, and Dethe S. Effect of Cinnamomum cassia methanol extract and
sildenafil on arginase and sexual function of young male Wistar rats. J Sex Med 2014;11:1475–1483.
Key Words. Cinnamomum cassia; Arginase; Enzymes and Enzyme Inhibitors Regulating Corporal Smooth Muscle
Relaxation; Sexual Function; Sildenafil; NOHA
Introduction
Penile erection is an integral part of male
sexual function, and many neurotransmitters,
ion channels, and enzymes are involved in the
control of erectile mechanisms, centrally and
peripherally [1]. Cyclic guanosine monophos-
phate (cGMP)-specific phosphodiesterase type 5
inhibitors (PDE5is) like sildenafil, vardenafil, and
tadalafil are commonly used for the management
of erectile dysfunction (ED), a common male
sexual dysfunction [2,3]. PDE5is act by relaxing
corpus cavernosum smooth muscle (CCSM)
(penile tissue smooth muscle) and increasing
penile blood flow. Other enzymes and their
inhibitors, which have been implicated in the
management of ED, include arginase [4] and Rho
kinase 2 (ROCK-II) [5].
Indian medicinal plants described as Vajikaran
herbs/aphrodisiacs have been used for the man-
agement of male sexual disorders including ED
[6–11]. Because of a lack of literature, it is difficult
to comment on the mechanism of action of these
plants. However, the efficacy of these extracts
1475
© 2014 International Society for Sexual Medicine J Sex Med 2014;11:1475–1483
might be attributed to inhibition of one or more
of the above mentioned enzymes. One such
plant with an unknown mechanism of action is
Cinnamomum cassia.
Methanol extract of the C. cassia (CCME) was
previously reported by authors of this article to
inhibit ROCK-II in vitro with an IC50 of
9.40 ±1.93 μg/mL [12]. In this study, we evaluated
the effect of CCME on in vitro arginase activity
and isolated rat CCSM. Effect of the extract was
also studied on sexual function of young male rats
in addition to its effect on smooth muscle : colla-
gen level in penile tissue of these rats.
Materials and Methods
Plant Material and Extraction
Bark of C. cassia (500 g) was procured from a local
market in Bangalore, India, during January 2011.
The raw material was identified and authenticated
by comparison with the crude drug reference stan-
dard by Dr. P. Santhan, Taxonomist, Natural Rem-
edies Pvt. Ltd. The voucher specimen was stored
in the natural product library of Natural Remedies
Pvt. Ltd., and methanol extract of remaining dried
bark was prepared as described elsewhere [12].
Chemicals and Materials
L-Arginine, MnCl2,N
ω-hydroxy-L-arginine
(NOHA, an arginase inhibitor), and Sirius red
(direct red 80) from Sigma-Aldrich, Co. (St. Louis,
MO, USA), dimethylaminobenzaldehyde, bovine
serum albumin (BSA), potassium sodium tartrate,
picric acid, xylene, and Tween 20 from HiMedia
Labs (Mumbai, India), and Folin–Ciocalteu
reagent from Sisco Research Laboratories Pvt.
Ltd. (Mumbai, India) were collected. Diethylstil-
bestrol from Penta Pharmaceuticals (Mumbai,
India) and progesterone from Sun Pharmaceutical
Ind. Ltd. (Mumbai, India) were procured. Watson
Pharma India Ltd. (Mumbai, India) provided
sildenafil as a gift sample. All other reagents used
in the study were of analytical grade.
Animals
The animal experimentation was performed after
review and approval of the study protocol by our
institutional animal ethics committee. Young
Wistar rats of either sex weighing 200–250 g were
fed with normal rat chow, had free access to drink-
ing water, and were maintained in 12-hour light
and dark cycles at 25°C.
Isolation of Arginase Enzyme
Crude arginase enzyme was isolated from liver of
rat as described by the method of Schimke [13]
with some modifications. Briefly, rat weighing
250 g was anaesthetized and sacrificed by cervical
dislocation. Thoracic cavity was opened up and
liver was carefully removed. The liver was washed
twice in 0.01 M Tris-HCl buffer containing
0.05 M MnCl2(pH 7.5). Further, liver was cut into
pieces and homogenized in three volumes of ice-
cold buffer. The homogenate was centrifuged at
4°C, 15,000 gfor 15 minutes, and supernatant was
separated. To the supernatant, 1.5 volumes of
chilled acetone (−10°C) was added, mixed, and
centrifuged at −10°C, 15,000 gfor 5 minutes. The
precipitate was homogenized in five volumes of
buffer and centrifuged at 4°C, 15,000 gfor 10
minutes. The supernatant was collected and dia-
lyzed for 20 minutes against buffer in an ice bath
using a magnetic stirrer. The dialyzed solution was
heated at 60°C for 20 minutes on water bath while
continuously stirring it with glass rod. The result-
ing solution was cooled on ice bath and centri-
fuged again at 4°C, 15,000 gfor 10 minutes. The
supernatant was cooled to 0°C followed by addi-
tion of equal volume of chilled ethanol (−10°C)
containing 0.05 M MnCl2. The mixture was cen-
trifuged at −10°C, 15,000 gfor 10 minutes. Three
volumes of ethanol (4°C) were added to the above
supernatant and centrifuged at 4°C, 15,000 gfor
10 minutes. The precipitate was finally suspended
and homogenized in 3 mL of 0.01 M Tris-HCl
buffer containing 0.05 M MnCl2(pH 7.5) and
stored at −80°C till use.
Arginase Inhibition Assay
Protein content of crude arginase enzyme was esti-
mated by the method of Lowry et al. using BSA as
a standard [14].
In arginase (inhibition) assay, specific activity of
arginase can be determined by the amount of argi-
nine consumed or the amount of urea formed. We
measured the amount of urea generated in argi-
nase (inhibition) assay to establish the specific
activity of arginase used. Absorbance of urea-
Ehlrich’s reagent (p-dimethylaminobenzaldehyde
in 3.6 N H2SO4) complex of the assay was plotted
against the standard curve of different concentra-
tion of urea and a fixed concentration of Ehlrich’s
reagent [15].
Arginase inhibition assay was performed as per
the method of Hagan and Dallam [16]. In brief,
a solution containing crude enzyme (protein
1476 Goswami et al.
J Sex Med 2014;11:1475–1483
concentration of 30 μg/mL) and 50 μg/mL of
herbal extract were pre-incubated for 5 minutes in
0.01 mM Tris-HCl buffer (pH 7.5) containing
0.05 mM MnCl2. The reaction was initiated by
addition of 50 mM L-arginine followed by incu-
bation at 37°C for 10 minutes. The reaction was
stopped by an addition of 4 ×10−4M Ehlrich’s
reagent, and the microplate was read at an absor-
bance of 450 nm using a microplate reader
(VERSAmax; Molecular Devices, Sunnyvale, CA,
USA) after 20 minutes. The plant extract was pre-
pared in 50% dimethyl sulfoxide (DMSO),
wherein final concentration of DMSO in the assay
was limited to 2.5%. NOHA was used as positive
control in this study. IC50 was calculated and the
study was performed in triplicate.
Similarly, arginase inhibition potential of
sildenafil at 50 μg/mL was also studied.
Effect on Isolated Rat CCSM
As described previously [12], two corpus
cavernosa (3 ×3×15 mm) were isolated from
each young male Wistar rats weighing 200–230 g,
anesthetized by intraperitoneal administration of
ketamine (70 mg/kg) and xylazine (10 mg/kg).
Tunica albuginea covering corpus cavernosum was
partially removed, and the CCSMs were mounted
in a four-channel organ bath containing modified
Krebs–Henseleit (K-H) salt solution (composition
[mM]: 118 NaCl, 4.7 KCl, 1.2 KH2PO4, 1.2
MgSO4, 1.5 CaCl2, 25 NaHCO3, 11 glucose, 0.01
Na2EDTA, and 0.006 indomethacin) maintained
under standard conditions (temperature: 37°C;
aeration with carbogen gas; replacing K-H solu-
tion every 15 minutes). Tissues were stretched up
to 500 mg of tension by a steel hook at the bottom
of organ bath and a force transducer (MLT0201;
ADInstruments, Bella Vista, NSW, Australia)
positioned at the top of organ bath, and connected
to PowerLab/8SP data acquisition system (Chart
7.0; ADInstruments). After equilibration for 1
hour, relaxant effects of DMSO (2.5%), CCME,
or sildenafil were recorded separately on phenyl-
ephrine (3 μM) pre-contracted rat CCSM.
Percent relaxation of CCSM by the vehicle,
extract, and sildenafil was calculated considering
maximum contraction of tissue by phenylephrine
as 100% response.
Sexual Behavior Study
Sexually experienced young (5 months, weighing
200–250 g) male Wistar albino rats were used for
the study. Rats were divided into three groups with
each containing six rats: group I (young rat treated
with 1% Tween 20), group II (young rat treated
with 100 mg/kg body weight CCME), and group
III (young rat treated with 5 mg/kg body weight
sildenafil). Treatment was continued for 28 days
and sexual behavior of male rats was observed on 0
(before dosing), 14th, and 28th day of dosing
[7,8,10].
Ovariectomized [17] female rats were brought
to estrus phase by administration of diethylstil-
bestrol (1 mg/kg, p.o., administered 48 hours
prior to study) and progesterone (5 mg/kg, s.c.,
administered 4 hours prior to the study). One
hour after dosing, male rat was introduced to the
observation chamber (45 ×50 ×35 cm) illumi-
nated by red light and allowed to accustom to the
environment for 5 minutes. Female rat in estrus
phase was then introduced into the cage by lifting
upper glass lid slowly, and a number of sexual
behavior parameters were observed. Mount
latency (ML) (time taken by the male rat up to
the first mount on female rat) and intromission
latency (IL) (time taken by the male rat up to the
first intromission) were recorded. Ejaculation
latency (EL) (time taken by the male rat from
first intromission until ejaculation) was confirmed
by presence of semen on vagina of female rat or
vaginal plug. Mount frequency (MF) (number of
mounting by a male rat on a female rat in a
given sexual cycle), intromission frequency (IF)
(number of intromission by a male rat in a given
sexual cycle), and post-ejaculatory interval (PEI)
(time from end of one sexual cycle to start of
another consecutive sexual cycle) were also
studied. The PEI was characterized by lethargy
of the male rats.
Histopathology of Rat Penile Tissue
Penile tissues were isolated from anesthetized rats,
and histopathology of the tissues was performed as
described earlier [18,19] with modification.
Briefly, the excised shaft of the penile tissues were
fixed over night in 7% formalin saline and then
transferred to 70% ethanol till the tissues were
further processed. The tissues samples were dehy-
drated in graded ethanol for 30 minutes, followed
by clearing in chloroform twice for 30 minutes
each. The tissues were then impregnated in paraf-
fin block and cut into five micron thick in size for
fixing to glass slides.
For determining smooth muscle : collagen level
in rat penile tissues, Masson’s trichrome staining
was performed. The tissue were hydrated and
dipped in Coplin staining jar containing Bouin’s
Cinnamon Increases Sexual Function of Young Male Rats 1477
J Sex Med 2014;11:1475–1483
fluid for half a day at 27°C. The tissues were
washed and rinsed with distilled water. The tissues
were stained in Weigert’s iron hematoxylin
(10 minutes) and Biebrich scarlet-acid fuchsin
staining (5 minutes). The tissues were stained
with aniline blue (3 minutes) after treating in
phosphomolybdic-tungstic acid solution. The
tissues were rinsed in dilute acetic acid (5 minutes).
Tissues were washed and rinsed with distilled
water between different staining procedures.
Tissues were then dehydrated in alcohol, cleared
in xylene, and mounted in dibutyl phathalate
xylene resin. Tissue slides were studied under a
light microscope (40×; Nikon, Otawara, Japan)
and photographs of CCSM were taken with a
Nikon digital camera (model number: DS-Ri1).
Statistical Methods
Inhibition of enzyme activity by herbal extract
and sildenafil is expressed as mean ±standard
deviation.
Effects of vehicle, herbal extract, and sildenafil
on isolated rat CCSM, and sexual behavior are
expressed as mean ±standard error of mean. Sta-
tistical significance with respect to vehicle was
evaluated using one-way analysis of variance fol-
lowed by Dunnett’s t-test using SPSS software
version 17 (SPSS Inc., Chicago, IL, USA).
Results
Arginase Inhibition Assay
As shown in Figure 1, CCME was found to be a
more potent arginase inhibitor than NOHA [20].
IC50 of CCME and NOHA was found to be
61.72 ±2.20 and 156.78 ±7.76 μg/mL, respec-
tively. Specific activity of arginase was found to be
490 μM urea/minute/mg of protein. Protein con-
centration of arginase stock solution (crude
enzyme) was found to be 3.068 mg/mL, and
30 μg/mL of crude enzyme was used for assay.
Sildenafil (a PDE5-specific inhibitor) did not
inhibit in vitro arginase activity up to 250 μg/mL.
Effect on Isolated Rat CCSM
Percent tissue relaxation by CCME and sildenafil
was studied at four different concentrations i.e.,
0.1, 1, 10, and 100 μg/mL. Both CCME and
sildenafil relaxed CCSM at these concentrations in
a dose-dependent manner. At highest concentra-
tion, sildenafil relaxed CCSM beyond 100% while
CCME showed up to 43% relaxation (Figure 2).
Sexual Behavior Study
Sexual behavior increased significantly in young
Wistar rats after C. cassia and sildenafil treatment
as is evident by the decrease in ML, IL, and PEI
and increase in MF, IF, and EL. ML and PEI are
measures of sexual motivation whereas intromis-
sion depends on relaxation of penile smooth
muscle [1,21]. Treatment with CCME decreased
ML, IL, and PEI in young male rats while the
treatment increased MF, IF, and EL. The treat-
ment increased sexual function of rats in a time-
dependent manner. Effect of sildenafil was better
than CCME (Figure 3).
80
60
Cinnamomum cassia ME
NOHA
0.79
1.04
1.33
1.69
1.74
log concentration (mg)
2
2.04
2.34
40
Percent Arginase inhibition
20
0
Figure 1 Effect of Cinnamomum cassia methanol extract
and NOHA on arginase inhibition in vitro. ME =methanol
extract; NOHA =Nω-hydroxy-L-arginine.
Figure 2 Relaxant effect of Cinnamomum cassia (n =5)
and sildenafil (n =6) on isolated rat corpus cavernosum
smooth muscle (CCSM) vs. vehicle (DMSO, n =6). The
relaxant effect of C. cassia methanol extract and sildenafil
was recorded at 0.1, 1, 10, and 100 μg/mL and compared
with that of DMSO. Each bar represents mean ±standard
error of mean. *P<0.001. DMSO =dimethyl sulfoxide.
1478 Goswami et al.
J Sex Med 2014;11:1475–1483
A
B
C
Figure 3 Increase in sexual behavior
of young rats treated with methanol
extract of Cinnamomum cassia
(CCME) and sildenafil (SIL) vs. vehicle
(V). n =6. 0, 14, and 28 denotes
day(s). Values are expressed as
mean ±standard error of mean.
#P≤0.05, $P≤0.01, *P≤0.001.
EL =ejaculation latency; IF =intromis-
sion frequency; IL =intromission
latency; MF =mount frequency;
ML =mount latency; PEI =post-
ejaculatory interval.
Cinnamon Increases Sexual Function of Young Male Rats 1479
J Sex Med 2014;11:1475–1483
Histopathology of Rat Penile Tissue
Treatment with CCME and sildenafil increased
the level of smooth muscle and decreased the level
of collagen in the rat penile tissue (Figure 4).
Discussion
In our previous study, we have reported ROCK-II
inhibition potential of CCME [12]. This study
further revealed arginase inhibition potential of the
extract. CCME was more potent than NOHA, an
endogenous arginase inhibitor. Pro-erectile effect
of C. cassia was proved as it significantly relaxed
(P<0.001) phenylephrine-contracted, isolated rat
CCSM. CCME also decreased ML and PEI and
increased MF, IF and EL significantly. The increase
in sexual function [21] justifies the role of CCME in
Vajikaran chikitsa, a branch of Ayurveda for man-
agement of male sexual dysfunction [6].
Arginine/nitric oxide (NO)/cGMP pathway was
found to relax CCSM, which is essential for penile
erection [22]. Up-regulation of arginase in the
penis of aged animal has been reported to deplete
arginine level resulting in decreased erectile func-
tion [23]. L-arginine 5 g per day for 6 weeks
increased sexual function of human participants
[24]. Further, the relaxation of phenylephrine-
contracted, isolated human corpus cavernosum has
also been shown to increase after incubation with
L-arginine for 6 to 10 hours [25]. Neuronal nitric
oxide synthase (nNOS) utilizes arginine for pro-
duction of NO that diffuses into CCSM and
stimulates soluble guanylyl cyclase to generate
cGMP from guanosine triphosphate. cGMP, the
second messenger, relaxes the penile smooth
muscle by opening potassium channel. Arginase
competes with nNOS for arginine and decreases
the level of cGMP, and arginase inhibition
increases cGMP level in penile smooth muscle
(Figure 5).
Rho kinase was reported to be present in human,
rabbit, and rat CCSM and its inhibition by Y-27632
dihydrochloride has been reported to relax CCSM
[26,27]. In another study, Y-27632 improved sexual
function in aged rat where elevated RhoA/Rho-
kinase activity had decreased intracavernosal pres-
sure [28]. Recently, Toque et al. demonstrated that
ROCK-II is associated with diabetes-induced
increase in the level of arginase in mice penile
smooth muscle. Corpora cavernosa (CC) from dia-
betic mice showed reduced relaxation response to
acetylcholine in comparison with CC from normal
mice, whereas CC from Rho-kinase knockout dia-
betic mice showed better relaxation response. Argi-
nase level and activity were found to be increased in
the CC of diabetic mice, whereas CC from Rho-
kinase knockout diabetic mice had reduced arginase
level and activity [29]. Waldkirch et al. demon-
strated the presence of Rho-kinase-related proteins
in human cavernous arteries. Y-27632 was reported
to relax norepinephrine pre-contracted cavernous
arteries. This study confirmed the role of Rho-
kinase in penile blood flow and endothelial relax-
ation [30]. The erectile potential of apocynin
(nicotinamide adenine dinucleotide phosphate
oxidase inhibitor), a phytochemical in diabetic rats,
was reported to be due to Rho-kinase inhibition
[31]. Because the increase in the amount of dephos-
Figure 4 Microscopic image of rat corpus cavernosum smooth muscle (CCSM) treated with vehicle/methanol extract of
Cinnamomum cassia (CCME)/sildenafil stained with Masson’s trichrome staining and imaged through light microscopy (40×).
Smooth muscle (shown by arrows) was stained as red/pink, while collagen (shown by broken arrow) was stained as blue.
Treatment with CCME and sildenafil increased smooth muscle and decreased collagen level.
1480 Goswami et al.
J Sex Med 2014;11:1475–1483
phorylated myosin light chain as a result of
ROCK-II inhibition leads to relaxation of isolated
rat CCSM, CCME-induced relaxation can be
partly attributed to ROCK-II inhibition.
The relaxation of rat CCSM and increase in IF
by CCME might be due to stimulation of
arginine/NO/cGMP pathway and inhibition of
RhoA/Rho-kinase pathway.
The level of smooth muscle was greater in the
corpus cavernosum of rats treated with CCME
when compared with rats treated with vehicle.
This increase in smooth muscle level by CCME
could be due to its cinnamic acid content. Ethanol
extract of C. cassia and cinnamic acid (a marker
compound for C. cassia) were reported to induce
endothelial cell proliferation in vitro. The extract
and cinnamic acid also increased the production of
vascular endothelial growth factor (VEGF) and
fetal liver kinase-1/kinase insert domain receptor,
the VEGF receptor involved in proliferation of
endothelial cells [32]. cGMP has been reported to
possess antifibrotic property [18]. Decrease of col-
lagen level in penile tissue of rats treated with
CCME and sildenafil could be due to increase in
level of cGMP due to arginase and PDE5 inhibi-
tion, respectively.
Conclusion
Erectogenic and aphrodisiac effects of CCME bark
can be attributed to its potential to inhibit arginase
activity and increase smooth muscle collagen ratio
in the young rat penile tissue. Thus, the study
supports aphrodisiac use of C. cassia in Ayurveda as
sexual invigorator and suggests CCME as a poten-
tial lead for the management of ED. Further studies
should be conducted to know the efficacy of C. cassia
in age and diabetes-induced ED.
Acknowledgments
This study was supported by research grant (R&D/KR-
03/2009–10-NMPB) to Al-Ameen College of Phar-
CCSM
GTP
cGMP
sGC NO
nNOS
Ornithine
+ Urea
Arginase
Relax
CCME
PDE5
GMP
Sildenafil
L-Arginine
Cavernous nerve
Figure 5 A proposed mechanism of Cinnamomum cassia action for increasing corpus cavernosal smooth muscle function.
Arrow represents stimulation, whereas dotted arrow represents inhibition. Arginine is a common substrate for nNOS and
arginase for production of NO and urea, respectively. NO after diffusing to CCSM from nonadrenergic noncholinergic nerve
terminal (cavernous nerve) stimulates sGC for converting GTP to cGMP that relaxes CCSM. Inhibition of arginase increases
the level of arginine that ultimately leads to more production of cGMP. However, PDE5 can metabolize cGMP, decreasing
relaxant effect of cGMP. Sildenafil relaxes CCSM by inhibiting PDE5. Both C. cassia and sildenafil works through arginine/
NO/cGMP pathway, but inhibit different enzymes. CCME =methanol extract of Cinnamomum cassia; CCSM =corpus
cavernosum smooth muscle; cGMP =cyclic guanosine monophosphate; GTP =guanosine triphosphate; nNOS =neuronal
nitric oxide synthase; NO =nitric oxide; PDE5 =phosphodiesterase type 5; sGC =soluble guanylyl cyclase
Cinnamon Increases Sexual Function of Young Male Rats 1481
J Sex Med 2014;11:1475–1483
macy, Bangalore and Natural Remedies Private
Limited, Bangalore from National Medicinal Plants
Board, New Delhi. We are thankful to Mr. Deepak
Kumar Khajuria for ovariectomy.
Corresponding Author: Sumanta Kumar Goswami,
PhD, Department of Pharmacology, Al-Ameen College
of Pharmacy, Near Lalbagh Main gate, Hosur road,
Bangalore 560027, India. Tel: +91-080-2211-3861; Fax:
+91-080-2222-5834; E-mail: sumantag@gmail.com
Conflict of Interest: The authors report no conflicts of
interest.
Statement of Authorship
Category 1
(a) Conception and Design
Sumanta K. Goswami; Rohitash Jamwal; Shekhar
Dethe; Mohammed N. Inamdar
(b) Acquisition of Data
Sumanta K. Goswami; Rohitash Jamwal
(c) Analysis and Interpretation of Data
Sumanta K. Goswami; Rohitash Jamwal; Shekhar
Dethe; Mohammed N. Inamdar
Category 2
(a) Drafting the Article
Sumanta K. Goswami
(b) Revising It for Intellectual Content
Rohitash Jamwal; Shekhar Dethe; Mohammed N.
Inamdar
Category 3
(a) Final Approval of the Completed Article
Sumanta K. Goswami; Rohitash Jamwal; Shekhar
Dethe; Mohammed N. Inamdar
References
1 Andersson KE. Pharmacology of penile erection. Pharmacol
Rev 2001;53:417–50.
2 Burchardt M, Burchardt T, Anastasiadis AG, Kiss AJ, Shabsigh
A, de La Taille A, Pawar RV, Baer L, Shabsigh R. Erectile
dysfunction is a marker for cardiovascular complications and
psychological functioning in men with hypertension. Int J
Impot Res 2001;13:276–81.
3 Carson CC. Erectile dysfunction: Evaluation and new treat-
ment options. Psychosom Med 2004;66:664–71.
4 Christianson DW. Arginase: Structure, mechanism, and physi-
ological role arginase in male and female sexual arousal. Acc
Chem Res 2005;38:191–201.
5 Wingard CJ, Johnson JA, Holmes A, Prikosh A. Improved
erectile function after Rho-kinase inhibition in a rat castrate
model of erectile dysfunction. Am J Physiol Regul Integr
Comp Physiol 2003;284:R1572–9.
6 Dalal PK, Tripathi A, Gupta SK. Vajikarana: Treatment of
sexual dysfunctions based on Indian concepts. Indian J Psy-
chiatry 2013;55:S273–6.
7 Thakur M, Chauhan NS, Bhargava S, Dixit VK. A compara-
tive study on aphrodisiac activity of some Ayurvedic herbs in
male albino rats. Arch Sex Behav 2009;38:1009–15.
8 Tajuddin, Ahmad S, Latif A, Qasmi IA. Aphrodisiac activity of
50% ethanol extracts of Myristica fragrans Houtt. (nutmeg) and
Syzygium aromaticum (L) Merr. & Perry. (clove) in male mice: A
comparative study. BMC Complement Altern Med 2003;3:6.
9 Chauhan NS, Rao ChV, Dixit VK. Effect of Curculigo orchioides
rhizomes on sexual behaviour of male rats. Fitoterapia
2007;78:530–4.
10 Suresh S, Prithiviraj E, Prakash S. Dose- and time-dependent
effects of ethanolic extract of Mucuna pruriens Linn. seed on
sexual behavior of normal male rats. J Ethnopharmacol
2009;122:497–501.
11 Gauthaman K, Adaikan PG, Prasad RN. Aphrodisiac proper-
ties of Tribulus terrestris extract (Protodioscin) in normal and
castrated rats. Life Sci 2002;71:1385–96.
12 Goswami SK, Pandre MK, Jamwal R, Dethe S, Agarwal A,
Inamdar MN. Screening for Rho-kinase 2 inhibitory potential
of Indian medicinal plants used in management of erectile
dysfunction. J Ethnopharmacol 2012;144:483–9.
13 Schimke RT. The importance of both synthesis and degrada-
tion in the control of arginase levels in rat liver. J Biol Chem
1964;239:3808–17.
14 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein
measurement with the folin phenol reagent. J Biol Chem
1951;193:265–75.
15 Hirsch-Kolb H, Greenberg DM. Molecular characteristics of
rat liver arginase. J Biol Chem 1968;243:6123–9.
16 Hagan JJ, Dallam RD. Measurement of arginase activity. Anal
Biochem 1968;22:518–24.
17 Parhizkar S, Ibrahim R, Latiff LA. Incision choice in
laparatomy: A comparison of two incision techniques in ovari-
ectomy of rats. World Appl Sci J 2008;4:537–40.
18 Ferrini MG, Kovanecz I, Sanchez S, Vernet D, Davila HH,
Rajfer J, Gonzalez-Cadavid NF. Long-term continuous treat-
ment with sildenafil ameliorates aging-related erectile dysfunc-
tion and the underlying corporal fibrosis in the rat. Biol
Reprod 2007;76:915–23.
19 Goswami SK, Inamdar MN, Pandre MK, Jamwal R, Dethe S.
Erectogenic and aphrodisiac effects of Butea frondosa Koenig ex
Roxb. in rats: Involvement of enzyme inhibition. Evid Based
Complement Alternat Med 2013;2013:874894. doi: 10.1155/
2013/874894.
20 Daghigh F, Fukuto JM, Ash DE. Inhibition of rat liver argi-
nase by an intermediate in NO biosynthesis, NG-Hydroxy-
L-Arginine: Implications for the regulation of nitric oxide
biosynthesis by arginase. Biochem Biophys Res Commun
1994;202:174–80.
21 Agmo A. Male rat sexual behavior. Brain Res Protoc
1997;1:203–9.
22 Trigo-Rocha F, Aronson WJ, Hohenfellner M, Ignarro LJ,
Rajfer J, Lue TF. Nitric oxide and cGMP: Mediators of pelvic
nerve-stimulated erection in dogs. Am J Physiol 1993;264:
H419–22.
23 Bivalacqua TJ, Burnett AL, Hellstrom WJG, Champion HC.
Over expression of arginase in the aged mouse penis impairs
erectile function and decreases eNOS activity: Influence of in
vivo gene therapy of anti-arginase. Am J Physiol Heart Circ
Physiol 2007;292:H1340–51.
24 Chen J, Wollman Y, Chernichovsky T, Iaina A, Sofer M,
Matzkin H. Effect of oral administration of high-dose nitric
oxide donor L-arginine in men with organic erectile dysfunc-
tion: Results of a double-blind, randomized, placebo-
controlled study. BJU Int 1999;83:269–73.
25 Gur S, Kadowitz PJ, Trost L, Hellstrom WJ. Optimizing nitric
oxide production by time dependent L-arginine administration
in isolated human corpus cavernosum. J Urol 2007;178:1543–8.
1482 Goswami et al.
J Sex Med 2014;11:1475–1483
26 Wingard CJ, Johnson JA, Holmes A, Prikosh A. Improved
erectile function after Rho-kinase inhibition in a rat castrate
model of erectile dysfunction. Am J Physiol Regul Integr
Comp Physiol 2003;284:R1572–9.
27 Rees RW, Ziessen T, Ralph DJ, Kell P, Moncada S, Cellek S.
Human and rabbit cavernosal smooth muscle cells express
Rho-kinase. Int J Impot Res 2002;14:1–7.
28 Jin L, Liu T, Lagoda GA, Champion HC, Bivalacqua TJ,
Burnett AL. Elevated RhoA/Rho-kinase activity in the aged rat
penis: Mechanism for age-associated erectile dysfunction.
FASEB J 2006;20:536–8.
29 Toque HA, Nunes KP, Yao L, Liao JK, Webb RC, Caldwell RB,
Caldwell RW. Activated Rho kinase mediates diabetes-induced
elevation of vascular arginase activation and contributes to
impaired corpora cavernosa relaxation: Possible involvement of
p38 MAPK activation. J Sex Med 2013;10:1502–15.
30 Waldkirch ES, Ückert S, Sohn M, Kuczyk MA, Hedlund P.
Rho kinase (ROK)-related proteins in human cavernous arter-
ies: An immunohistochemical and functional approach. J Sex
Med 2012;9:1337–43.
31 Li M, Zhuan L, Wang T, Rao K, Yang J, Yang J, Quan W, Liu
J, Ye Z. Apocynin improves erectile function in diabetic rats
through regulation of NADPH oxidase expression. J Sex Med
2012;9:3041–50.
32 Choi DY, Baek YH, Huh JE, Ko JM, Woo H, Lee JD, Park
DS. Stimulatory effect of Cinnamomum cassia and cinnamic
acid on angiogenesis through up-regulation of VEGF and Flk-
1/KDR expression. Int Immunopharmacol 2009;9:959–67.
Cinnamon Increases Sexual Function of Young Male Rats 1483
J Sex Med 2014;11:1475–1483
