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EDITORS:

Michael P. Ward (Australia)

Faouzi Kechrid (Africa)

Montip Gettayacamin (Thailand)

Fedik Abdul Rantam (lndonesia)

Suzanita Utama (lndonesiai

@

^ZOLZ

Center Publishing

and

Printing

of

Airlangga University (AUP)

AUP 600/1 6. 443 I 06.12-B2E

All

^rights reserved. No part of this book may be reprinted or reproduced or

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in any

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or

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an electronic, mechanical, or other means, including photocopying and recording, or in any information storage in retrieval system,

without

prior permissio,n in

writing

from the publisher.

First

print

-2012

Publisher:

Center Publishing and Printing

ofAirlangga University (AUP)

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Printed by: Center Publishing

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of

National

Cataloging-in-Publication

Data

Pro

Proceeding International Seminar: Strategy to Manage Bio-Eco-Health System

for

Stabilizing

Animal

Health and Productivity to Support Public Health/

Ed: Michael P. Ward

...let al.l

-

First Print

-Surabaya:

Center Publishing and Printing

ofAirlangga

University, 2012

lxxvi,342p.t2l ^x

29,7 cm

Bibliography

rsBN

978-602-8967 -69 -3

l.

Veterinary Public Health

I.

^Faouzi

^Kechrid

II. Montip

Gettayacamin

III. FedikAbdul

Rantam

lV.

Suzanita Utama

636.083 2

1213t4151619876s4321

MEMBER OF IKAPI: OO1,/JTI,/95

THE EFFECTS OF HYPERBARIC OXYGEN ON THE NUMBER ^OF' EOSINOPHILS AND THE PICTURES OF SPLEEN WHITE PULP

DIAMETERS IN WHITE RATS GIVEN HEAVY SWIMMING EXERCISES

Setianingsih, H.

Deparftnent of Anatomy and Histology

Faculty of Medicine Hang Tuah University, Surabaya, Indonesia E-mail : dr.xfanayy a@gmail. com

ABSTRACT

In the modern era today, there are many severe stressor such as the increase ofheavy physical exercises may lead individual susceptibility to either psychological or physical stress. Frequent and heavy physical exercise can affect the type and number of white blood cells in circulation expecially eosinophils (Nernet et al, 2004). ^Stress is one

of

immunosupressive factors that can lower the body immunity system, fertility and spur the emergence of free radicals, resulting in damage to cell membranes, causing inflammation and degenerative diseases. The use of hyperbaric oxygen (HBO) is a kind of medication in which a patient breathes 100% oxygens through a mask and is under a pressure more than

I ATA

(2.4 ATA)

in

a certain period. In this condition, the oxygen circulation in the body increases 2-3 times bigger than the normal condition (Thom, 1989). Hlperbaric oxygen has been used to fix a tissue, which experiences inflammation, ischemia and endotel disorders, whereas HBO will spurthe formation ofnew endotels. However, there hasnlbeen a research ofthe effect ofhyperbaric oxygen to immunity system that involves eosinophils and spleen. Therefore, this research aims to know the effect of HBO 2.4 ATA to the number of eosinophils and the pictures of spleen white pulp diameter in white rats given heavy swimming exercises. This research used 30 white rats (Rattus norvegicus wistar strain), a chamber of HBO and oxygen. The parameter of this research used the number of eosinophils and spleen white pulp diameters. Experimental animals used in this study were 30 white rats who were randomly divided into one control group and two treatment groups. K0

:

control group, treatment group

K:

^{1 (heavy swimming exercises} for eight hours everyday in four weeks). The treatment group K2

:2

(heavy swimming exercises for eight hours everyday in four weeks ar,d2.4

AIA

hyperbaric oxygen therapy for 90 minutes. The given hyperbaric oxygen therapy 2.4 NIA for 90 minutes to the rats with heavy swimming exercises

will

decrease the number of egsinophils and maintain the spleen white pulp diameter.

The results ofthis iesearch support the previous research that found the increase ofoxidative stress during anaerobic exercises. Moreover the increase ofoxidative stress also leads the increase ofprotein and fats are oxidized ultimately lead to mutations of cells, damage the immune system and tissues, including spleen (Groussard et a1.,2003). Immune system is determined by some factors, including eosinophils (Cruse and Lewis, 2003) prepared by the reticuloendothelial tissues, bone marrow, spleen, lymph node that serves to fight infections and form anti-bodies. Research on the effect of HBO on the immune response is also not yet widely applied. Physical exercise can indicate changes in the secretion ofthe hormone in the pituitary gland, thyroid, pancreas, adrenal and sex (Bullock and Henze, 2000). One ofthe key response neuro- hormonal in physical exercise is an increase simpato-adrenal activity (Bouchard, 1993). Response changes are closely related to the needs ofthe body's metabolic processes and homeostasis. Once physical exercise can lead to changes in stress hormones that include an increase in AGTH, cortisol, enkepalin, thyroxine _, catecholamines and the decrease in anabolic hormones (insulin and testosterone) (Bullock and Henze, 2000). Physical exercise is a form ofphysical stress can increase the secretion ofvarious hormones (cortisol, endorphins, catecholamines) that serves as an immunomodulator (Mackinnon, 1992). The use of hyperbaric oxygen in rats with heavy swimming exercise can decrease in the number of eosinophils and maintain the spleen white pulp diameters.

Keywords: spleen white pulp, eosinophil, hyperbaric oxygen, exercise

INTRODUCTION

Under conditions of

stress is

often found

an increased

incidence of allergies in individuals,

even

with no history of allergies though.

(Parsons and

Mastronade, 2005)

says

that the exercise-induced bronchoconstriction (EIB)

experienced

by l0% of

^subjects

with

no

history of allergies

or asthma.

It is said that the

stress

of physical activity will result in an increase in eosinophils beginning

^and

86

then decreases

slowly

due to the

rapid expenditure of

adrenal

cortex hormones

(Shephard, 1982).

In

stress

conditions there is

some

level of

increase

of eosinophils in the study (Chatterjea

et a1.,1953) and a

maximum

increase

of eosinophils

ranged

from

9

to I20 minutes.

(Pedersen and

Goetz, 2000)

states

that

exercise

is

one

of

the

models

are

representative of physical

stress. Base

on

she research

(Susan, 2005)

suggests

that the stressors ofphysical exercise can increase the number ofblood eosinophils will migrate into the tissue causing an allergic reaction. Hyperbaric oxygen therapy

usage

and clinical applications for

several

health problems,

such as:

Gangrene Diabetes Mellitus, Neurological Disorders/Stroke, Otolaryngologis

cases, cases

ofbone

surgery, and

infection in burns

(Jain

KK,

2001).

Groussardet

al. have made

it clearthat

an increase

in oxidative

stress occurs

during strenuous physical exercise/anaerobic. This exercise will affect the immune system is played by

several factors

including leukocyte

cells.

Leukocytes

are also called

white blood

cells that have a

type

that includes phagocytosis

polymorfonuclear

neutrophils, eosinophils, and basophils, and

lymphocytes

and

monocytes

(Cruse and

Lewis,

2003) was made

by reticulo endothelial

tissues, red bone

marrow,

spleen,

lymph

node

that

serves

to counter germs

and

form anti-bodies. Up to

date

information on the number of eosinophils

and

white pulp

spleen

diameter image

on

white rats

exposed

to

stress is

still very limited. Therefore, it is still very

necessary research

to find out about the decline of

the

immune

system

in

rats exposed

to

stress.

This study

aimed

to determine

the decrease

in

the

number

of

eosinophils

and

white pulp

spleen

diameterpictures

on

white

rats

exposedto

stress

through heavy swimming

exercises.

IUATHERIATS AND METHODS

The study was conducted

at

the Laboratory of Biochemistry Faculty of Medicine, University

of

HangTuah

Surabaya,

for

6

months, start in April

2011

to

September 2011.

Laboratory examination performed

at

the Laboratory Prodia, Jl. Diponegoro

107 Surabaya and

histopathological examination

at the

Laboratory

of

Pathology Navy hospital

dr.

Ramelan

Surabaya.

Implementation

Research

White

rats adapted for 2 weeks.

During

the maintenance fed

with

a standard

formulation. this

research used 30

white rats (Rattus norvegicus wistar strain),

a chamber

of HBO

and

oxygen. The parameter of this

research used

the number of eosinophils and

spleen

white pulp

diameters.

Experimental animals

used

in this

study

were

30

white

rats

who were randomly divided into

one

control

group and

two

treatment groups.

K0:control

group. The treatment group

K:1 (heavy swimming

^exercises

for eight hours everyday in four

^weeks).

The treatment group K2=2 (heavy swimming

^exercises

for

eight hours everyday

in four weeks

and2.4

AIA hyperbaric

oxygen

therapy for

90

minutes.

This

research uses

experimental methods. Data generated in the analysis using the Kruskall Wallis

test

with 95% significant

level.

RESUTTS AND DISCUSSIONS

Table 1. ^The

^average

number of eosinophils

CONTROL EXERCISE EXERCISE & HBO

No. eosinophils

^%

eosinophils % eosinophils ^%

l.

2.

J.

4.

5.

6

2 2 J 2 2 2

I

2 2 2 2

a -')

2

1

2 2 2 2 106

132 297

64

t04

192

58 192 453 108

248

426

56 82 152

86

r28

210

CONTROL EXERCISE _EXERCISE & HBO

No. eosinophils % eosinophils % eosinophils %

7.

8.

9.

10.

53 266 48 124

1

2

I

2

I

2 2 2

2 2 Ja

2 86

82 164 52

126 130

63 78

Table 2. The

statistics

results

The

number eosinophils Chi-Square

df

Asymp.

Sig.

16.54s 2 .000

a, b.

Kruskal Wallis Test

Grouping Variable: treatment

The results of Kruskal Wallis test on the table 2 shows the number 0.000 indicates there

are

significant differences in the number of eosinophils in the control of white rats, with exercise

exercise and

were given hyperbaric oxygen therapy.

Stress

can activate the hypothalmic pituitary adrenocortical axis (HPA-axis) that

secrete or

produce corticotropin releasing factor (CRF), which

serves

to increase adenocorticotropin hormone (ACTH)

and

then

secreted

into the blood

vessels

throughout. Corticoadrenal tissue is responsible for the synthesis

and

release of ACTH with

an increase

in

steroid

hormones. The final result of hormonal activity in rats

characterized

by

elevated

hormone corticosterone and cortisol in the blood

are

classified

as

glucocorticoids

(Irlaseem

e/ al

2005). Continuous

release

of glucocorticoids which would

cause

atropy in lymphoid

organs such as

thymus,

spleen,

lymph

nodes.

It

can also

occur

due to

hyperbaric

oxygen encourages the

formation of

fibroblast

and increase the effects of leucocyte phagocytosis

loanandjaja, 200e. rhe

results showed

that administration of hyperbaric oxygen

is able

to maintain

the

diameter of

the

white pulp atrophy lien that

does not happen because

ofstressor

severe

intensity

exercise.

Table 3.

The mean

diameter of white pulp Lien

The

treatment

groups

KO

KI

K2

0.01525

0.0t224

0.01s20

Based

on

the

results of Kruskall Wallis

test

with 95% significant level (p < 0.05)

showed there

were significant differences in the diameter of the

splenic

white pulp in

the

iat with the

exercise

of

control

rats and rats

that

exercise and were

given hyperbaric

oxygen therapy.

An increasing number of glucocorticoids also

causes

inhibition of working interleukin-l (IL-li and interleukin-2 (lLe)

that play

a

role in the proliferation of T lymphocytes, B and B cell maturation into plasma cells, resulting in

the

formation

of antibodies to be

obstructed. Where

is the _spleen

functions

as a

producer of antibodies by B lymphocytes following antigen

challenged

(Firdaus, 2008)When T lymphocytes and B lymphocytes cell proliferation

due

to

a

decline in the number of glucocorticoid increased the number of T lymphocytes

and

B lymphocytes

10

in the white pulp also

decreased.

Therefore

contributes to the diameter of the

white pulp

that come

shrink

oratrophy.

Evident in

Table 3 show

that

the

diameter

of the spleen

white pulp

on average

in

the

control group (Ko)

0.01525

+

0.0049

mm

and treatment groups

(Kl)

0.01224 + _0.0253

mm.

Then the treatment group were given

hyperbaric oxygen therapy (K2)

0.01520

+

0.0055

mm. When

compared between the

control group (Ko)

and

treatment

88

r.MHERE,rr."o,ro,[;i?ly#f Hffi ffiIi,EJiffi 'S;*ffi'.-i@@

(Kl)

a decrease

in diameter

of

white pulp,

where the

control group (Ko)

have on average

higher than

the treatment group but later

inthe

group

givenhyperbaric oxygentherapy

shows the average

number of almost the

same as

the control group, it is proved that the administration of hyperbaric oxygen in

rats

white heavy swimming

exercise can

maintain the diameter of the white pulp lien. This

can

occur

because

of

the use

of

the pressure

in hyperbaric oxygen therapy will

decrease

the volume of

gas bubbles and the use

of hyperbaric oxygen will

also accelerate the

resolution of the

gas bubbles

(Kindwall,

2008).

coNctusroN

Based on theresults ofdata analysisanddiscussionit canbe concludedthat the use

of

2.4 ATA

hyperbaric oxygen therapy for 90 minutes

in white

rats

with

heavy

swimming

exercises

for

eight hours everyday

in four

weeks,

will

reduce the

number

of

blood eosinophils

and can

maintain

the

picture of

spleen

white pulp diameter significantly.

R.EFERENCES

Bouchard C. 1993. Physical

Activity,

Fitness, and Health Consensus Statement: Physical

Activity,

Fitness and Disease. 1" ed. Canada: Human

Kinetic

Publisher. P.

6l-82.

Bullock BL

and Henze

RL.

2000. Focus On Patophysiology. Lippincott

Williams

and

Wilkins.

Chatterjea JB, Dameshek

W

and

Stefanini M.

1953. The

Adrenaline (Epinephrine)

Test as

Applied

to Hematologic Disorder. Blood 8:

p.2ll-235.

Cruse JM, Lewis RE. Atlas of immunology. USA: CRC Press, Inc; 2003.

Danandjaja, Danny Wresniwira, 2000. Pengaruh Pemberian Hiperbarik Hiperoksi

Dan Kcndisffipotatk

Terhadap Respon Imun Seluler Prajurit

Marinir. Tesis.

Universitas Airlangga.Surabaya

Firdaus S.D., 2008. Enhancing versus suppressive effects

of

stress on

immune function: implication fbr

immunoprotection versus immunopathology.

Allergy, Asthma

and

Clinical Immunology.

Stanford University, Standford. 4: p,

2-ll.

Groussard C, Rannou-Bekono F, Machefer G, Chevanne M, Vincent S,Sergent O, Cillard

I

Gratas-Delamarche

A

(2003). Changes in blood lipid peroxidation markers and antioxidants after a single sprint anaerobic exercise, Europ. J. Appl. Physiol.89: 14-20.

Jain

K K,

2009. Physical, physiolqgical and biochemical aspect

of hyperbaric

oxygenation. Textbook

of

Hyperbaric Medicine. 3rd ed. Hogrefe and Huber Pub. Pp: 10-25.

Kindwall

E P, 2008. The physiologiq effect of hyperbaric oxygen. Hyperbaric Medicine Practice. 2nd ed. Best Publishing Company, USA. Fp:

17J2.

Mackinnon

Ul.

1992. Exercise and Immunology. Humen Kinetics Books, Canada.

Naseem, M.T., Shamoon Naseem,

M.

Yunus; Zafar lqbal Ch.,

Aamir

Ghafoor,Asim Aslam and S. Akhter.

2005. Effect of Pottasium Choride and.Sodium Bicarbonate Supplementation on Thermotolerance

of

Broiler Exposed to Heat Stress. International Journal Of Poultry Science 4 (11): 891-895.

Nernet D,

Mills

PJ and Cooper

DM.

2004. Effect of intense wrestling exercise on leucocytes and adhesion molecules in adolescent boys. Br J Sports Med 38: pp 154-158.

Parsons JP and

Mastronaide

JG, 2005. Exercise-Induced

Bronchoconstriction in Athletes. Chest

128:

p.3966-3974.

Pedersen

BK

and

Hoffman-GoetzL.2000.

Exercise and the immune system: Regulationl Integration and Adaptation. Phys Rev 80: pp 1055-1081.

Shephard RJ, 1980. Physiology and Biochemistry of Exercise, USA: Praeger Publisher: p.306.

Susan

L,

Cuvelie SL, Paul S, Shariat

N,

Colarusso P and Patel

KD,

2005. Eosinophil adhesion under

flow

conditions activates mechanosensitive signaling pathways

in

human endothelial cells.

JEM

202 (6), pp.865-876.

Thom SR., 1989. Hyperbaric oxygen therapy. J Int Care Med 4:

5814.

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