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Vijay Kumar Chavan*et al. /International Journal of Pharmacy & Technology

IJPT| Sep-2017| Vol. 9 | Issue No.3 | 30742-30762 Page 30742

ISSN: 0975-766X

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www.ijptonline.com OPTICAL MICROSCOPIC ANALYSIS ON SAMPLES OF ALBINO MICE FED WITH SILDENAFIL

CITRATE AND ALCOHOL

Vijay Kumar Chavan*1, M. Sreenivasulu

2

1,2Dept of Science & Hiumanities, VFSTR University, Guntur, A.P., India.

Email: [email protected]

Received on: 21-08-2017 Accepted on: 25-09-2017

Abstract:

Erectile dysfunction (ED), a common disorder among men, is on the surge in recent days. Even though many

treatments are available, Sildenafil citrate is proven to be the best oral treatment. Sildenafil is an oral

phosphodiesterase inhibitor that enhances erectile function through the same general pathway used by nature.

Sildenafil also has been shown to be effective in men with hypertension, diabetes, nonvascular organic etiologies for

ED, and psychogenic causes. Sildenafil citrate and ethanol consumption are used in societies worls wide and have

been idenfied as injurious to human health. The aim of this study determine changes in Histoarchitecture of Heart and

Testis of Albino mice treated combinedly with Caverta and Ethanol of using microscopic technique. Healthy male

Wistar Albino mice (Mus Musculus), aged about 60 days and weighing about 25-30 gm, were procured from central

Animals House. Animals were selected and grouped into seven each group consisting of six animals (Group S1, S2,

S3, S4, S5, S6 and S7). Drug, here, refers to the 50mg tablet of Sildenafil Citrate (CAVERTA Ranbaxy, India)

dissolved in an appropriate Volume of Conductivity water to get 20ppm solution. This solution was used as the drug

and the animals were fed with this drug @ 1µg/gm body weight. Ethanol, here, represents 18% or 100ppm of Ethanol

and the experimental Albino Mice were fed with this Ethanol solution @ 0.01 µg/gm body weight. The animals in the

group S1 were considered as control animals and were fed intragastrically with conductivity water. The animals in the

group S2 and S3 were fed with a single close of the drug chosen (Caverta) for 15days and 30 days respectively. The

groups S4 and S5 were treated with a single close of Ethanol for 15 and 30 days respectively and the animals

belonging to the last pair of groups (S6 and S7) were treated with a single close of drug and Ethanol combinedly for

15 and 30 days respectively. The animals belonging to the groups S2, S4 and S6 were sacrificed on 15th

day of initial

drug administration while those belonging to S3, S5 and S7 were sacrificed on 30th

day of initial drug administration.

All these experimental animals were decapitated on the terminal day of the dosage, after four hours of drug

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administration. The operative procedures were carried under strict aseptic precautions. A vertical ventral midline

incision was made in the abdominal wall to collect both the left and right testis and Heart samples, after using open

Ketlar anaesthesia. These collected testis samples were examined by using Optical Microscopic technique.. The long

term treatment Ethanol fed Albino mice with Caverta (30days) the Testis showed a total distortion in the

Histoarchitecture of the seminiferous tubules involving vacuolization of cells, prominent Intersitial Oedema and

various congestion. The outcome is drug – induced made severe morphological changes in the tissue of Testis of

experimental Albino mice. The changes in the architecture of Heart samples were found in all experimental groups.

The adverse effects on the normal architecture of Heart such as the loss of normal branching pattern of the cardiac

muscle fibre, loss of myocytes, venous congestion, Odema and vacuolated cells were detected in the case of long

term drug treated Heart samples of Albino mice fed combinedly with Sildenafil citrate and Ethanol.Therefore, it is

concluded that the combined dosage of Caverta and Ethanol produce adverse effects on Heart and Testis of Albino

mice. It is suggested that the utilization of sophisticated analytical techniques would probably throw limelight on the

mechanism of action of these drugs.

Keywords: Albino mice, Erectile Dysfunction, Ethanol, Heart, , Intersitial Oedema, Morphological studies,

Sildenafil citrate (Caverta), Testis, .

Introduction

In worlds momentum the role of human being plays vital role. These human beings with different attitudes are living

in the society. Some are taking alcohol, drugs and most of the people taking both drug and alcohol, for their personal

entertainment and sexual purpose. But very few people in the world under controle i.e., they are with perfect diet and

nutrition.. But maximum people they habituated in taking alcohol and silidenifilcitrate or Cialis or Caverta or Viagra

tabltes for enhancing their extra pleasure during their sex. But this entertainment may cause damage to the orgasms of

human being. To Prove this the author taken albino mice and he has fed Ehanol and siledenifil Citrate as mentioned

in methodology part.Albino mice are most drug sensitive animals and Laboratory Mammals and most important

mode organisms in Medicine and Biology. It is also belonging to strains of House Mice and is easy to handle,

maintain, reproduce quickly and share a high degree of homology with humans. Inability to attain and maintain an

erection sufficient to permit satisfactory sexual intercourse is called Erectile dysfunction (ED)(NIH Consensus

Development Panel on Impotence, NIH Consensus Conference, 1993). By survey it is found that ED was present

in 671 (52%) Massachusetts men Out of 1290, aged 40 to 70 year. (Feldman et al., 1994). 67% patients with


IJPT| Sep-2017| Vol. 9 | Issue No.3 | 30742-30762 0744

coronary artery disease reported that within a average span of 3 years their ED symptoms began before their

coronary artery disease symptoms (Montorsi et al., 2003). As the general way used in nature Sildenafil is an oral

Phosphodiesterase inhibitor that enhances erectile function [(Goldstein et al., 1998) amd (Kloner et al., 1999))].

For ED, and psychogenic causes Sildenafil shown to be effective in men with hypertension, diabetes, nonvascular

organic etiologies (Goldstein et al., 1998). Ethanol is considered a psychoactive drug to change the human

consciousness and it is a central nervous system depressant and has significant psychoactive effects in sublethal

doses ( David et al., 2002). A blood level of 0.5% or more is commonly fatal. Levels of even less than 0.1% can

cause intoxication, with unconsciousness often occurring at 0.3 – 0.4%amd also may cause death for alcohol

consumption people (Hingson et al., 2003).

Aim & Objective

To determine changes in Histoarchitecture of Heart and Testis of Albino mice treated combinedly with Caverta and

Ethanol by using microscopic technique.

Materials and Methods

Animals

Healthy male Wistar Albino mice (Mus Musculus), aged about 60 days and weighing about 25-30 gm, were procured

from central Animal House, Rajah Muthaiah Medical College, Annamalai University, Annamalai Nagar, Tamilnadu.

These animals were fed with standard pellet diet (Hindustan Lever Ltd., Mumbai, India) and water ad libitum.

Grouping

Animals were selected and grouped into seven, each group consisting of six animals (Group S1, S2, S3, S4, S5, S6 and

S7).

Drug Treatment

Drug, here, refers to the 50 mg tablet of Sildenafil Citrate (CAVERTA-Ranbaxy, India) dissolved in an appropriate

volume of conductivity water to get 20 ppm solution. This solution was used as the drug and the animals were fed with

this drug @ 1µg/gm body weight. Ethanol, here, represents 18% or 100 ppm of Ethanol and the experimental Albino

Mice were fed with this Ethanol solution @ 0.01 µg/gm body weight.

Dosage: The animals in the group S1 were considered as control animals and were fed intragastrically with

conductivity water. The animals in the group S2 and S3 were fed with a single dose of the drug chosen (Caverta) for

15 days and 30 days respectively.

23 23

http://en.wikipedia.org/wiki/Psychoactive

http://en.wikipedia.org/wiki/Drug

http://en.wikipedia.org/wiki/Human_consciousness



http://en.wikipedia.org/wiki/Ethanol#cite_note-79

http://en.wikipedia.org/wiki/Alcohol_intoxication



The groups S4 and S5 were treated with a single dose of Ethanol for 15 and 30 days respectively and the animals

belonging to the last pair of groups (S6 and S7) were treated with a single dose of drug and Ethanol combinedly for 15

and 30 days respectively.

The animals belonging to the groups S2, S4 and S6 were sacrificed on 15th day of initial drug administration while

those belonging to S3, S5 and S7 were sacrificed on 30th day of initial drug administration. All these experimental

animals were decapitated on the terminal day of the dosage, after four hours of drug administration.

Principle of an Optical Microscope

A Microscope is in principle a simple lens system for magnifying small objects. The first lens called the

Objective has a short focal length (a few mm), and creates an image of the object in the intermediate image plane.

This image, in turn, can be looked at with another lens, the eye - piece, which can provide further magnification .The

resolution of the image is limited by diffraction.

The Abbe Rayleigh Criterion states that, for a wavelength λ, the smallest distance dmin resolvable between two point

sources as deduced from diffraction theory is

dmin = 1.22 x λ/2NA

Where NA = n x Sinα is called Numerical aperture of the Objective lens.

n = index of refraction in the object space.

α= half the maximal angle under which the Objectives lens collects light

from the object.

Sample Preparation

The Heart and Testis tissues collected from both the control and the experimental animals were fixed using Bowin’s

fluid and then dehydrated using alcohol. After dehydration, they were cleared using xylene and embedded in a

Paraffin wax. Separate paraffin blocks for each tissue were prepared. Using a Rotary Microtome (Weswox Company,

India), sections of 7 µm thickness were cut.

These sections were deparaffinised in xylene and the slides were initially stained with Haemotoxylin and then with

Eosin. After staining, these slides were dehydrated through ascending grades of alcohol, cleared in xylene and

mounted in DPX. Using the procedures described above, each Testis sample was serially sectioned and a

minimum of 100 sections per Testis sample were stained and studied. Morphological studies were made using the

stage microscope (NIKON, Japan).



Results and Discussion

Morphological Studies of Testis

The Testis samples of control and experimental animals have been collected and subjected to Morphological studies

using Light Microscopic technique with an intention of studying the drug - induced structural changes

Fig.1.1 to Fig. 1.11 represent the section of Testis samples of control and experimental Albino mice. Following

observations have been made :

Interpretation of Testis Slides

In case of S1 (0 hr.) samples, the Testis of Albino mice show normal pattern consisting of interstitial space and

Basement membrane.

The structure of the Testis was intact as shown in Fig.1.1 and Fig 1.2.

After fifteen days of continuous dosage of the drug [Sildenafil citrate (Caverta)] treatment, the Testis of the

experimental Albino mice exhibited mild Interstitial Oedema and separation of spermatogonia cells from Basement

membrane (Fig.1.3).

After a single dose of Caverta treatment continuously for 30 days (S3), there is an enhancement in Interstitial Oedema

and venous congestion. Separation of spermatogonia cells from basement membrane was more and fat bubbles were

also observed (Fig.1.4 and Fig.1.5).

In the case of Testis of Albino mice samples (S4) treated with Ethanol for 15 days continuously @ 0.01 µg/g body

wt., a normal texture of seminiferous tubule, Interstitial space and mild separation of spermatogonia cells from

basement membrane were observed (Fig 1.6).

As the duration of Ethanol treatment gets increased to 30 days (S5), there was mild interstitial Oedema and mild

separation of cells from basement membrane as shown in Fig.1.7.

In the case of S6 samples, treated with the combined dosage of Caverta and Ethanol continuously for 15 days, a

noticeable increase in the cells at the centre of the seminiferous tubule and enhancement of Interstitial Oedema along

with distinct separation of spermatogonia cells from the basement membrane were detected and are evident from

Fig.1.8 and Fig.1.9.

The long term treatment of Ethanol fed Albino mice with Caverta (S7 – 30 days) resulted in the Testis exhibiting a

total distortion in the Histoarchitecture of the seminiferous tubules involving vacuolization of cells, prominent

Interstitial Oedema and venous congestion (Fig. 1.10 and Fig 1.11).



Slides of testis:

Fig.1.1 Section of Testis of Albino mice [S1(0 hr.)] showing normal pattern consisting of (1) seminiferous

tubule, (2) Interstitial space, (3) basement membrane and (4) Lumen. Haematoxylin – eosin x100

Fig.1.2 Section of Testis of Albino mice [S1 (0 hr.)] showing normal pattern consisting of (1) Interstitial space

and (2) Basement membrane. Haematoxylin – eosin x100.

Fig1.3 Section of Testis of Albino mice [S2(15 days Caverta treated)] showing (1) Interstitial Oedema and (2)

separation of Spermatogonia cells from Basement membrane. Haematoxylin – eosin x40



Fig.1.4 Section of Testis of Albino mice [S3(30 days Caverta treated )] showing (1) marked Interstitial

Oedema, (2) Venous congestion and (3) marked separation of Spermatogonia cells from Basement

membrane. Haematoxylin – eosin x40.

Fig.1.5 Section of Testis of Albino mice [S3 (30 days Caverta treated)] showing (1) venous congestion and (2)

marked separation of Spermatogonia cells from Basement membrane and (3) fat bubble .

Haematoxylin – eosin x100.

Fig.1.6 Section of Testis of Albino mice [S4 (15 days Ethanol treated)] showing (1) Seminiferous tubule, (2)

Interstitial space and (3) mild separation of spermatogonia cells from Basement membrane.

Haematoxylin - eosin x40.



Fig.1.7 Section of Testis of Albino mice [S5 (30 days Ethanol treated)] showing (1) mild Interstitial Oedema

and (2) mild separation of cells from Basement membrane. Haematoxylin – eosin x40.

Fig.1.8 Section of Testis of Albino mice [S6 (15 days combined dosage of Caverta and Ethanol)] showing (1)

Interstitial Oedema, (2) Separation of cells from Basement membrane and (3) increased number of cells at the

centre of the Seminiferous tubule. Haematoxylin – eosin x40.


marked Interstitial Oedema, (2) noticeable increase in the cells at the centre of the Seminiferous

tubule and (3) marked separation of cells from the Basement membrane. Haematoxylin – eosin x100.


striking alterations in the Histoarchitecture of the Seminiferous tubule. Haematoxylin –

eosin x40.



Fig.111 Section of Testis of Albino mice [S7 (30 days combined dosage of Caverta and Ethanol)] showing (1)

marked alterations in the Histoarchitecture of the Seminiferous tubule and (2) Vacoulization of cells in the

Seminiferous tubule. Haematoxylin– eosin x100.

Discussion on Testis

The results obtained from the present research work indicate the fact that the long term co-administration of Caverta

and Ethanol will produce drastic morphological changes such as distorted Histoarchitecture, venous congestion,

prominent Interstitial Oedema and vacuolization of cells in Testis of the experimental Albino mice. These observed

results can be discussed in detail as follows:

Anatomical pattern and temporal order of testicular development and it makes cell differentiation, proliferation,

apoptosis and migration(Capel, 2000).However, the loss of body weight and testicular weight, reduced diameter of

the seminiferous tubules, early separation of spermatogonia cells and prominent Interstitial Oedema were reported for

Testis samples of long term Sildenafil citrate (Viagra) treated Albino mice (Suriyakumari, 2003) and Albino rats

(Savitha, 2006). In the present investigation also, similar Testicular morphological changes have been noticed for

Sildenafil citrate (Caverta) treated animals.

Chronic Ethanol effects human and animals testicular function. It also reduces testis atrophy, testosterone

production,reduced and sperm output[(Van Thiel et al., 1980), (Adler, 1992) and (Villalta et al., 1997)]. Decrease

in diameter of seminiferous tubules causes loss of germ cells in Chronic alcoholics Testes (Van Thiel et al., 1975).

Germ cell apoptosis can be increased animals by Ethanol exposure(Zhu et al., 2000) and also secretory function of

Sertoli cells may cause adverse effect (Zhu et al., 1997). In adult rats apoptosis of germ cells can be increased by

Ethanol [(Zhu et al., 2000) and(Eid et al., 2002)]. Immature germ cells apoptosis can be enhanced by Ethanol and

this enhance may cause abnormal spermatogenesis in adults during neonatal period due to in first 3–5 postnatal days

normal development of germ cells is critcal[(Orth et al., 1988)and (de Rooij, 1998)].Based on the above discussion,



it is quite clear that the Histological changes such as Interstitial Oedema and separation of Spermatogonia cells from

the basement membrane due to long term Ethanol exposure, as detected in the present study, are the strong indication

of the negative impact of Ethanol on Testis of Albino mice.

Similar morphological changes , as reported in the present work, have been detected in the case of Testis samples

treated with environmental chemicals. Estrogenic property in animals and humans may cause many reproductive

disorders due to environmental chemicals both natural and artificial (Chitra et al., 1999).. Specific causes of

testicular damage have been catalogued in Testes of humans and other mammals due to environment or chemical

exposure and genetic disorders(Jadaramkunti and Kaliwal, 2002). In last 50 years as per the Epidemiological

studies in males the quality and quantity of sperm has been decreased due environmental agents and

drugs[(Bendvold et al., 1991) and (Carlsen et al., 1992)].

The physical and chemical agents in which precursor cells form mature haploid spermatozoa within seminiferous

tubules and spermatogenesis is a highly sensitive process. The fall of sperm count and infertility in wildlife and

human due to the overproduction of reactive oxygen species (ROS) in both intra and extra cellular

spaces(Gangadharan et al., 2001).

Pesticide like other organophosphorus is largely used as Phosphamidonand it makes toxic effects to the pest by

phosphorulation and accumulation of acetylcholine is caused due to active centre of acetylcholineesterase(Eto,

1974). Male reproductive system if exposed chronically is found to be toxic due toPhosphamidon and also enhanced

interstitial space, Reduction in weight of testis, spermcells, germ cells, leydig cells and sertoli cells. In testis the

cytotoxic effects of phosphamidon like deformities in testes and cytoplasmic vacuolization of germ cells (Ram

Bahadur kuwar et al., 1996)

Albino rats treated with carbaryl found some degeneration changes in the ells of seminiferous tubular germinal layers

seminiferous tubular germinal layers [Rybakova (1966), Vashakidze (1975), Pant et al. (1995 & 1996) and Rani

Archana et al. (2007)] The exact mechanism for these changes are still not clear. The spermatogenesis was also

depressed which is well supported by the similar findings by Kitagawa (1977), who reported reduced number of

spermatogonia in the experimental group

The Testis is sensitive to a variety of stressors, such as hyperthermia, inflammation, radiation and exposure to agents

that induce apoptosis of germ cells [(Lue et al., 1999), (O’Bryan et al., 2000), (Hasegawa et al., 1997) and

(Richburg, 2000)]. Oxidative stress in the Testis is one of the major factors that induces germ cell apoptosis, Among

66



various phthalate esters, DEHP is one of the most widely studied toxicants in the male reproductive organs.

Administration of DEHP reduces the fertility and induces testicular atrophy of laboratory animals [(Oishi et al.,

1979) (Thomas et al., 1984) and (Oishi, 1986)]. Administration of DEHP induces vacuolation in Sertoli cells

[(Richburg et al., 1996) and (Emiko Kasahara et al., 2002)].

In all doses of treatment it is found that there is significant chnge in diameter of seminiferous tubules, diameter of

lumen and seminiferous epithelium height were decreased and these chnges due to tubules disorganization and

spermatogenic cell loss(Creemers et al., 2002).

Hafeiz et al. (1984) have reported that a single dose of fraction VII from L. quinquestriatus venom given to rats is

capable of inducing degenerative changes in seminiferous tubules, resulting in necrozoospermia. Action of Nigella

sativa seems to be like other several anti-oxidative antidotes (Vitamin E , Ascorbic acid & Melatonin) which prevent

the degeneration of male germ cell by minimizing testicular cytotoxic effect in animal treated with pesticides,

chemicals, mutagens, and metals [( Foraga, 1991), (El –Bahy, 1997) and (Hsu et al., 1998)].

The administration of combined dosage of Sildenfil citrate (Caverta) and Ethanal for a long term may induce

vacuolization of cells in the seminiferous tubule and drastic alterations in thtubule.

Morphological Studies of Heart

Heart samples were collected from control and the experimental Albino mice and subjected to Light microscopic

analysis..

Fig.2.1 to Fig.2.9 represent the slides of the section of Heart of control and experimental animals treated for different

duration of time.

Interpretation of Heart Slides

The Histological sectioning of Heart samples of group S1 (0 hr.) animals exhibited very clear and well defined normal

Histoarchitecture involving normal branching of cardiac muscle fibre, centrally placed nucleus and Intercalated disc

(Fig.2.1 and Fig.2.2).

The venous congestion, loss of cells and Oedema were observed in the Heart samples of Sildenafil citrate treated

Albino mice of group S2 (15 days) (Fig.2.3])

The Light microscopic sectioning of Heart of Sildenafil citrate fed Albino mice of Group S3 (30 days) portrayed

venous congestion, oedema and marked loss of cells prominently (Fig. 24).



Administration of Ethanol continuously for 15 days resulted in mild distortion of normal branching pattern of the

cardiac muscle fibre and Oedema (Fig.2.5).

In the case of Heart samples of Ethanol fed Albino mice of Group S5 (30 days), Oedema and cellular infiltration were

noticed (ig.2.6).

Fig. 2.7 represents the section of Heart samples of Albino mice treated combinedly with Sildenafil citrate and Ethanol

[S6 (15 days)]. In these samples venous congestion, cellular infiltration, loss of myocytes and Oedema were noticed.

However, loss of normal architecture along with venous congestion, cellular infiltration and Oedema were striking

noticeable in Heart samples of S7 (30 days) group of Albino mice treated with combined dosage of Sildenafil citrate

and Ethanol. Besides, the occurrence of vacuolated cells is the characteristic feature in these samples [Fig.2.8 &

Fig.2.9].

Fig. 2.1 Section of Heart of Albino mice [ S1 (0 hr.)] showing (1) normal branching pattern of cardiac

muscle fibre, (2) centrally placed nucleus and (3) Intercalated disc. Haematoxylin – eosin x100

Fig. 2.2 Section of Heart of Albino mice [S1 (0 hr.)] showing (1) centrally placed nucleus and (2)

Intercalated disc. Haematoxylin – eosin x400



Fig. 2.3 Section of Heart of Sildenafil citrate treated Albino mice [S2 (15 days)] showing (1) venous

congestion, (2) loss of cells and (3) Oedema. Haematoxylin – eosin x100

Fig.2.4 Section of Heart of Sildenafil citrate treated Albino mice [S3 (30 days)] showing (1) venous

congestion, (2) Oedema and (3) marked loss of cells. Haematoxylin – eosin x100

Fig.2.5 Section of Heart of Ethanol fed Albino mice [S4 (15 days)] showing (1) a mild distortion in the

normal branching pattern of the cardiac muscle fibre and (2) Oedema. Haematoxylin – eosin

x100

Fig.2.6 Section of Heart of Ethanol fed Albino mice [S5 (30 days)] showing (1) Oedema and (2) cellular

infiltration. Haematoxylin - eosin x100



Fig.2.7 Section of Heart of Albino mice treated with the combined dosage of Sildenafil citrate and

Ethanol [S6 (15 days)] showing (1) venous congestion, (2) cellular infiltration, (3) loss of myocytes and (4)

Oedema. Haematoxylin – eosin x100


Ethanol [S7 (30 days)] showing (1) venous congestion, (2) cellular infiltration, (3) Oedema and (4)

loss of normal architecture. Haematoxylin – eosin x100


Ethanol [S7 (30 days)] showing (1) vacuolated cells and (2) cellular infiltration. Haematoxylin

– eosin x400



Discussion on Heart

The Heart samples of Albino mice (Ganesan, 2006) and Albino rats (Ravikumar, 2006) treated, on long – term

basis, with Sildenafil citrate have been reported to exhibit the most interesting features like the loss of normal

branching of the cardiac muscle fibres, cellular infiltration, venous congestion loss of myocytes, oedema and in

severe cases, the vacuolization of cells also. They have attributed these morphological changes to Sildenafil citrate

intoxication. Cardiovascular deaths and retinal vascular events in individuals taking Sildenafil have also been

reported [(Corbin et al., 2000) and (Cunningham et al., 2001)]. Vatansever et al. (2003) have concluded that the

chronic use of Sildenafil citrate would result in retinal Oedema.

Heavy consumption of alcohol also appears to affect Heart muscle and possibly arterial tissues directly (Doll et al.,

1994). Intercalary discs describes About cellular theory of Heart muscle. These discs are adjacent plasma membranes

separated by an interspace while the sarcolemma appears as plasma membrane, interspace plus basement membrane

of the interstitial. By way of endoplasmic reticulum the nucleus of the cell is closely associated with the entire cell

by way of endoplasmic reticulum (Dan et al., 1956).

Changes in rat cardiac muscle following a Magnesium – deficient regime and cold stress were studied, histologically

and by electron microscope, by Alexander et al. (1964). Focal myocardial necrosis, calcification and fibrosis

occurred in the Mg deficient animals. These changes were aggravated by cold stress and inhibited by Magnesium

supplements. Ultrastructural alterations included swelling and vacuolization of sarcosomes and fragmentation of

myofibrils. The sarcosomal abnormalilties appeared to be fundamental to the pathogenesis of the lesions.

The link between stress and acute myocardial infarction was found to be via Catecholamine induced intravascular

platelet thrombosis ( Jacob et al., 1972). A study was made by Ananya et al. (2005), to explore the effect of oral

administration of Lindane on Lipid peroxidation and histopathological changes of rat Hearts. Interstitial Oedema in

the myocardium was observed along with ultrastructural changes such as loss of integrity of myofibrils, Z- band

disruption and mitochondrial damage. Lipid peroxidation of the Heart as measured by Thiobarbituric acid reactive

substances (TBARS) was increased.Oxidative stress plays a key role in the pathogenesis and progression of major

cardiovascular disorders.

Follis et al. (1942) studied rats on a diet deficient in potassium and found constant changes in the myocardium. After

eight days on the diet, multiple small areas of muscle necrosis appeared. Concurrently, these areas were infiltrated

first by polymorphonuclear leucocytes and later by mononuclear cells.



In Histological study made by Karunakaran et al. (2006), Oedema, focal haemorrhage and leukocyte infiltration

were observed in Heart samples of Isoproterenol fed rat. In a study made by Karthikeyan et al. (2002), Albino rats

were subjected to chronic administration of Isoproterenol which led to myocardial injury. A significant rise in

myocardial Thiobarbutiric acid reactive substances (TBARS) and loss of reduced Glutathione (GSH), Superoxide

dismutase (SOD) and Catalase (suggestive of increased oxidative stress) occurred in the rat Hearts.

Chronic administration of Isoproterenol (ISP) to Wistar Albino rats has been reported to induce focal myonecrosis

with myophagcytosis and lymphocytic infiltration, vacuolar changes such as intracytoplasmic vacuoles and Lipid

droplets as well as prominent Oedema in Heart (Ojha et al., 2008). Rathore et al. (1998) and Blasig et al. (1984)

have emphasised that free radical mediated peroxidation of membrane permeability were the primary factor for ISP

induced Cardiotoxicity.

Therefore, Albino mice were fed with the combined dosage of Sildenafil citrate and Ethanol, on a long – term basis,

Ultrastructural changes such as mitochondrial damage and loss of integrity of myofibrils and thus, deleterious effects

on the structure and function of Heart may occur.

CONCLUSION

The long term treatment of Albino mice with combined dosage of Sildenafil citrate (Caverta) and Ethanol would

adversely affect the form and vital functions of Testis. And Heart.

Conflict of interest No conflict of interest was declared.

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Corresponding Author:

Vijay Kumar Chavan*,

Email: [email protected]

mailto:[email protected]

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