ABSTRACT

EFFECT OF CHRONIC SYSTEMIC HYPEROXIA TO

CONCENTRATION OF MALONDIALDEHYDE IN BLOOD AND
BRAIN OF SPARGUE DAWLEY RAT

Neurodegeneratif disease is a major health problem in some countries. Hyperoxia

plays role in the pathogenesis of neurodegeneratif through out the mechanism of
oxidative stress. The consequence of hyperoxia is an increase the formation of
reactive oxygen species (ROS), which causes an imbalance of pro-oxidant and
antioxidant, causing damage to brain cells. The damage of the brain cells is caused
by lipid peroxidation of cells membrane and produce compounds called
malondialdehyde (MDA). MDA are clearly detectable in the blood and brain, so it
become one of the stress oxidative biomarker.
In this study, rats were divided into 5 group (n = 6 per group). Each group were
exposed to hyperoxia (75% O2) for 1 day, 3 days, 7 days, 14 days, while the
control group was given normoxia and measurement of the MDA, in addition to
the examination of blood gas parameters, hematology and histopathology. .
MDA concentration in the blood and brain increased significantly compared to
control group (p < 0,05). In the statistical test, there is a strong positive correlation
between MDA in blood and brain (Pearson, p=0,002, r2= 0,9805). In the blood gas
parameters there are respiratory acidosis and histopathology damage of the brain
cells.
It was concluded that chronic systemic hyperoxia causes oxidative stress, which
increases concentration of MDA in the blood and brain. This is supported by
changes in the blood gas parameters, hematology and histopathology.

Keywords: Chronic systemic hyperoxia, malondialdehyde (MDA), reactive

oxygen species (ROS).

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 ABSTRAK

PENGARUH HIPEROKSIA SISTEMIK KRONIK TERHADAP

KADAR MALONDIALDEHID PADA DARAH DAN OTAK TIKUS
SPARGUE DAWLEY

Penyakit neurodegeneratif merupakan masalah kesehatan utama di beberapa

negara. Hiperoksia berperan dalam patogenesis neurodegeneratif melalui
mekanisme stres oksidatif. Salah satu akibat dari hiperoksia adalah peningkatan
reaktif oksigen spesies (ROS) yang menyebabkan ketidakseimbangan pro-oksidan
dan antioksidan, sehingga menyebabkan kerusakan sel otak. Kerusakan sel otak
ini terjadi akibat reaksi peroksidasi lipid membran sel yang menghasilkan
senyawa malondialdehid (MDA) yang mudah terdeteksi di dalam darah dan otak,
sehingga MDA dapat berperan sebagai biomarker stres oksidatif.
Pada penelitian ini, tikus diinduksi hiperoksia sistemik kronik dimana tikus dibagi
menjadi 5 kelompok (n = 6 per kelompok) . Masing masing kelompok diberi
perlakuan hiperoksia (O2 75%) dalam sungkup hiperoksia selama 1 hari, 3 hari, 7
hari, 14 hari, sedangkan kelompok kontrol diberi normoksia dan dilakukan
pengukuran terhadap MDA, selain itu dilakukan pemeriksaan analisis gas
darah,hematologi dan histopatologi.
Kadar MDA darah maupun otak meningkat secara bermakna dibanding kelompok
kontrol (p<0,05). Pada uji statistik, terdapat kolerasi positif kuat antara kadar
MDA darah dan otak (Pearson, p=0,002 r2= 0,9805). Pada analisa gas darah
adanya asidosis respiratorik dan histopatologi terjadi kerusakan sel otak.
Disimpulkan bahwa hiperoksia sistemik kronik menyebabkan stres oksidatif yang
meningkatkan kadar MDA darah dan otak. Hal ini juga didukung dengan
perubahan parameter gas darah, hematologi dan histopatologi.

Kata-kata kunci: Hiperoksia sistemik kronik, malondialdehid (MDA), reaktif

oksigen spesies (ROS).

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 DAFTAR ISI

HALAMAN JUDUL........................................................................................i
HALAMAN PERNYATAAN ORISINALITAS...............................................ii
HALAMAN PENGESAHAN..........................................................................iii
KATA PENGANTAR.......................................................................................iv
HALAMAN PERNYATAAN PERSETUJUAN PUBLIKASI.........................v
ABSTRACT.....................................................................................................vi
ABSTRAK........................................................................................................vii
DAFTAR ISI.....................................................................................................viii
DAFTAR TABEL.............................................................................................xiv
DAFTAR GAMBAR........................................................................................xii
DAFTAR SINGKATAN...................................................................................xiii
DAFTAR LAMPIRAN.....................................................................................xiv
1. PENDAHULUAN.......................................................................................1
1.1 Latar Belakang......................................................................................1
1.2 Rumusan Masalah.................................................................................2
1.2.1 Pernyataan Masalah..............................................................................2
1.2.2 Pertanyaan Masalah..............................................................................2
1.3 Hipotesis Penelitian..............................................................................3
1.4 Tujuan Penelitian..................................................................................3
1.4.1 Tujuan Umum.......................................................................................3
1.4.2 Tujuan Khusus......................................................................................3
1.5 Manfaat Penelitian................................................................................3
2. TINJAUAN PUSTAKA.............................................................................4
2.1 Sistem Saraf Pusat.................................................................................4
2.2 Hiperoksia, Reactive Oxygen Species (ROS) dan Stres Oksidatif........4
2.3 Pengaruh Hiperoksia Terhadap Otak....................................................9
2.4 Pengaruh Hiperoksia Terhadap Malondialdehid...................................12
2.5 Kerangka Teori......................................................................................16
2.6 Kerangka Konsep..................................................................................17
3. METODOLOGI PENELITIAN................................................................18
3.1 Desain Penelitian..................................................................................18
3.2 Waktu dan Tempat Penelitian................................................................18
3.2.1 Waktu Pelaksanaan...............................................................................18
3.2.2 Tempat Pelaksanaan..............................................................................18
3.3 Populasi dan Sample Penelitian............................................................18
3.3.1 Populasi Penelitian................................................................................18
3.3.2 Sample Penelitian.................................................................................18
3.4 Penetapan Jumlah Hewan Coba...........................................................19
3.5 Kriteria Inklusi dan Eksklusi...............................................................19
3.6 Cara Kerja Penelitian...........................................................................20
3.6.1 Perlakuan Hiperoksia............................................................................20
3.6.2 Pembuatan Homogenat Otak dan Darah...............................................21
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3.6.3 Pengukuran Kadar Malondialdehid......................................................22
3.6.4 Analisa Gas Darah................................................................................24
3.6.5 Pemeriksaan Patologi Anatomi............................................................24
3.7 Keterangan Lolos Kaji Etik.................................................................24
3.8 Variabel Penelitian................................................................................24
3.8.1 Variabel Bebas......................................................................................24
3.8.2 Variabel Tergantung..............................................................................24
3.9 Definisi Operasional.............................................................................24
3.9.1 Hiperoksia.............................................................................................24
3.9.2 Malondialdehid.....................................................................................25
3.10 Instrumen Penelitian.............................................................................25
3.10.1 Bahan Penelitian...................................................................................25
3.10.2 Alat Penelitian.......................................................................................25
3.11 Pengumpulan Data................................................................................28
3.12 Analisis Data.........................................................................................28
3.13 Alur Penelitian......................................................................................29
3.14 Jadwal Penelitian..................................................................................30
4. HASIL PENELITIAN................................................................................31
4.1 Analisis Gas Darah dan Hematologi.....................................................31
4.2 Mikroskopik Otak Tikus.......................................................................36
4.3 Standar MDA........................................................................................40
4.4 Kadar MDA Darah................................................................................41
4.5 Kadar MDA Otak..................................................................................42
4.6 Perbandingan Kadar MDA Darah dan Otak.........................................43
5. PEMBAHASAN..........................................................................................44
5.1 Analisa Gas Darah dan Hematologi......................................................44
5.2 Kadar MDA Darah dan Otak................................................................45
5.3 Mikroskopik Otak Tikus Spargue dawley.............................................46
5.3 Keterbatasan Peneliti............................................................................46
6. KESIMPULAN DAN SARAN...................................................................47
6.1 Kesimpulan...........................................................................................47
6.2 Saran.....................................................................................................47
DAFTAR PUSTAKA......................................................................................48
LAMPIRAN....................................................................................................52
DAFTAR RIWAYAT HIDUP.........................................................................66

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 DAFTAR TABEL

Tabel 2.1 Pembagian Reactive Oxygen Species (ROS).................................5

Tabel 3.1 Uji TBA Metode Wills E.D............................................................23
Tabel 3.2 Jadwal Pelaksanaan Penelitan........................................................30
Tabel 4.1 Analisa Gas Darah dan Hematologi...............................................31
Tabel 4.2 Absorbansi Standar MDA..............................................................40
Tabel 4.3 Kadar MDA Darah.........................................................................41
Tabel 4.4 Kadar MDA Otak...........................................................................42

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 DAFTAR GAMBAR

Gambar 2.1 .. Proses Terjadinya Reactive Oxygen Species (ROS)...................6

Gambar 2.2 Tempat Pembentukan ROS........................................................7
Gambar 2.3 .. Proses Terjadinya Stres Oksidatif..............................................8
Gambar 2.4 .. Sumber, Efek dan Marker Stres Oksidatif.................................9
Gambar 2.5 Sawar Darah Otak......................................................................12
Gambar 2.6 .. Proses Peroksidasi Lipid............................................................13
Gambar 2.7. .Proses Pembentukan MDA........................................................15
Gambar 2.8 ..Kerangka Teori...........................................................................16
Gambar 2.9 . .Kerangka Konsep.......................................................................17
Gambar 3.1 Proses Pembedahan pada Tikus.................................................21
Gambar 3.2 Bahan dan Hasil MDA...............................................................23
Gambar 3.3 Homogenizer (tissue grinde)......................................................26
Gambar 3.4 .. Alat sentriguasi..........................................................................26
Gambar 3.5 Penangas Air..............................................................................27
Gambar 3.6 Vortex Mixer...............................................................................27
Gambar 3.7 Micropipette semiotomatik ependorf.........................................28
Gambar 3.8 ..Alur Penelitian...........................................................................29
Gambar 4.1 Pengaruh Hiperoksia Terhadap pO2...........................................32
Gambar 4.2 Pengaruh Hiperoksia Terhadap pCO2........................................32
Gambar 4.3 Pengaruh Hiperoksia Terhadap pH............................................33
Gambar 4.4 .. Pengaruh Hiperoksia Terhadap HCO3.......................................33
Gambar 4.5 Pengaruh Hiperoksia Terhadap Saturasi Oksigen......................34
Gambar 4.6 Pengaruh Hiperoksia Terhadap Hemoglobin.............................34
Gambar 4.7 Pengaruh Hiperoksia Terhadap Hematokrit...............................35
Gambar 4.8 Pengaruh Hiperoksia Terhadap Sel Darah Merah......................35
Gambar 4.9 . Mikroskopik otak tikus Sprague dawley...................................36
Gambar 4.10 Tanpa Perlakuan Perwarnaan HE............................................36
Gambar 4.11 Tanpa Perlakuan Perwarnaan PAS..........................................37
Gambar 4.12 Perlakuan Hiperoksia 14 hari - Perwarnaan HE .......................38
Gambar 4.13. Perlakuan Hiperoksia 14 hari - Perwarnaan PAS.......................39
Gambar 4.14 Kurva Standar MDA..................................................................40
Gambar 4.15 Kadar MDA Darah.....................................................................41
Gambar 4.16 Kadar MDA Otak.......................................................................42
Gambar 4.17 Perbandingan Peningkatan Kadar MDA Darah dan Otak..........43

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 DAFTAR SINGKATAN

1O2 Singlet Oksigen

4-HNE 5-Hydroxynoneal
8-OhdG 8-hydroxy-2' -deoxyguanosine
ADHD Attention Deficit Hyperactivity Disorder
ATP Adenosine Triphosphate
CA California
CO2 Karbondioksida
CU2 Tembaga
DNA Asam Deoksiribonukleat
Fe2 Besi
GABA Gamma Aminobutyric Acid
H* Hidrogren
Hb Hemoglobin
HE Hematoxilin Eosin
HHT asam 12-1-hydroxi-5,8,10 hepatadecatonoic
Ht Hematokrit
H2O Air
H2O2 Hidrogen Peroksida
HCO3 Asam Karbonat
L* Radikal Asam Lemak
LOO* Radikal Peroksi
LOOH Lipid Hidroperoksida
LH Rantai Asam Lemak
MDA Malondialdehid
NADP Nicotinamide Adenin Dinukleotida
NO Nitrik Oksida
NO2 Nitrik Dioksida
N2 Nitrogen
O2 Oksigen
O2- Anion Superoksida
OH Radikal Hidroksil
ONOO- Hidrogen Peroksida
PAS Periodic Acid Schiff
pCO2 Tekanan Karbondioksida
pO2 Tekanan Oksigen
PUFA Polyunsaturated fatty acid
PGG2 Prostaglandin Endoperosida hidroperoksi G2
PGH2 Prostaglandin Endoperoksida
RO- Radikal Alkoksil
ROO- Radikal Peroksil
ROS Reaktif Oksigen Spesies
SOD Superoksida Dismutase
SSP Sistem Saraf Pusat
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 TBA Thiobarbituric Acid
TCA Trichloroacetic Acid
TXA2 Tromboxan A2
SDM Sel Darah Merah
USA United State of America
UV Ultra Violet

DAFTAR LAMPIRAN

LAMPIRAN 1 Analisis Gas Darah dan Hematologi..........................................52

LAMPIRAN 2 Kadar MDA Darah dan Otak.....................................................55
LAMPIRAN 3 Kolerasi Antara Tekanan O2 Arteri dan Kadar MDA Darah.....57
LAMPIRAN 4 Kolerasi Antara Tekanan O2 Arteri dan Kadar MDA Otak.......59
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LAMPIRAN 5 Kolerasi Antara Kadar MDA Darah dan Otak...........................61
LAMPIRAN 6 Hasil Pemeriksaan Histopatologi...............................................63
LAMPIRAN 7 Keterangan Lolos Kaji Etik.......................................................65
LAMPIRAN 8 Daftar Riwayat Hidup................................................................66

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