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KARYA TULIS AKHIR
PENGARUH EKSTRAK BIJI COKLAT (Theobroma cacao) SEBAGAI
ANTIOKSIDAN TERHADAP REGRESI PLAK ATEROMA DINDING
ARKUS AORTA TIKUS PUTIH JANTAN (Rattus norvegicus strain wistar)
ATEROSKLEROTIK
Oleh:
SIDIKA YUNIA MUYASYARAHMA
201110330311017
UNIVERSITAS MUHAMMADIYAH MALANG
FAKULTAS KEDOKTERAN
2015
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LEMBAR PENGESAHAN
LAPORAN HASIL PENELITIAN
Telah disetujui sebagai hasil penelitian
untuk memenuhi persyaratan
Pendidikan Sarjana Fakultas Kedokteran
Universitas Muhammadiyah Malang
Tanggal 23 Januari 2015
Pembimbing I
Prof. Dr. dr. H. Djoni Djunaedi, Sp.PD., KPTI., FINASIM
Pembimbing II
dr. Yuliono Trika Nur Hasan, Sp.M
Mengetahui,
Dekan Fakultas Kedokteran
Universitas Muhammadiyah Malang
dr. Irma Suswati M.Kes.
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dr. Kusuma Andriana, Sp.OG Anggota
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KATA PENGANTAR
Segala puji bagi Allah SWT atas segala rahmat dan hidayah-Nya, shalawat
serta salam terlimpahkan kepada Nabi Muhammad SAW, keluarga dan para
sahabatnya. Syukur alhamdulillah, penulis telah berhasil menyelesaikan karya
tulis akhir yang berjudul “PENGARUH EKSTRAK BIJI COKLAT
(Theobroma cacao) SEBAGAI ANTIOKSIDAN TERHADAP REGRESI
PLAK ATEROMA DINDING ARKUS AORTA TIKUS PUTIH JANTAN
(Rattus norvegicus strain wistar) ATEROSKLEROTIK”.
Dalam penyelesaian karya tulis akhir ini penulis ingin mengucapkan
terima kasih yang tak terhingga kepada :
1. dr. Irma Suswati, M.Kes selaku Dekan Fakultas Kedokteran Universitas
Muhammadiyah Malang.
2. dr. Moch. Ma’roef, Sp.OG selaku Pembantu Dekan I Fakultas Kedokteran
UMM serta pembimbing I dalam karya tulis akhir ini.
3. dr. Rahayu, Sp.S selaku Pembantu Dekan II Fakultas Kedokteran UMM.
4. dr. Iwan Sys Indrawanto, Sp.KJ selaku Pembantu Dekan III Fakultas
Kedokteran UMM.
5. Prof. Dr. dr. H. Djoni Djunaedi, Sp.PD., KPTI., FINASIM selaku
pembimbing 1 atas bimbingan, dukungan dan saran yang telah diberikan
dalam penyusunan karya tulis akhir ini.
6. dr. Yuliono Trika Nur Hasan, Sp.M selaku pembimbing 2 atas bimbingan,
dukungan dan saran yang telah diberikan dalam penyusunan karya tulis akhir
ini.
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7. dr. Kusuma Andriana, Sp. OG selaku penguji atas bimbingan, dukungan dan
saran yang telah diberikan dalam penyusunan karya tulis akhir ini.
8. dr. Soebarkah Basuki, Sp.PA selaku pembimbing patologi anatomi atas
bimbingan, dukungan dan saran yang telah diberikan dalam penyusunan
karya tulis akhir ini.
9. Mbak Patmawati, Mas Nyono, Mas Miftah, Pak Joko dan segenap Asisten
Lab. Patologi Klinik Universitas Muhammadiyah Malang atas bimbingan,
dukungan, dan saran yang telah diberikan dalam penelitian.
10. Ayahanda Drs H. Nur Rahman, MM, Ibunda Hj. Puji Rahayu, S.Pd, serta
dr.Devy Yuspita Rahma dan Dinsa Okta Rahmawati yang selalu memberikan
kasih sayang, do’a, dukungan, semangat, kepercayaan dan segalanya demi
keberhasilan penulis.
11. Semua teman-teman (Firman, Ro’di, Galang, Wahyu, Priangga, Yusuf,
Brando, Fahmi, Faris, Aulia, Adhinda, Prastiti, Nada, Adhe, Pamela, dan
Dendy) yang telah memberikan bantuan dan dorongan baik moril maupun
materil sehingga terselesaikannya penyusunan tugas akhir ini dengan baik.
Karya tulis akhir ini masih jauh dari kesempurnaan, penulis mohon maaf
yang sebesar-besarnya dan mengharapkan saran dan kritik yang membangun.
Semoga karya tulis ini dapat menambah wawasan dan bermanfaat bagi semua
pihak.
Malang, Januari 2015
Penulis
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ABSTRAK
Muyasyarahma, Sidika Yunia. 2014. Pengaruh Ekstrak Biji Coklat (Theobroma
cacao) sebagai Antioksidan Terhadap Regresi Plak Ateroma Dinding Arkus
Aorta Tikus Putih Jantan (Rattus norvegicus strain wistar) Aterosklerotik.
Tugas akhir. Fakultas Kedokteran Universitas Muhammadiyah Malang.
Pembimbing: (1) Djoni Djunaedi* (2) Yuliono Trika Nur Hasan**
Latar Belakang: Penyakit jantung koroner (PJK) telah menjadi penyebab utama
kematian di dunia. Penyebab utama PJK adalah penyempitan arteri koronaria
besar di bagian proksimal akibat aterosklerosis. Pada aterosklerosis terjadinya
disfungsi endotel akibat terjadi penurunan fraksi antioksidan yang dapat
menstimulasi produksi Nitric Oxide (NO) yang berfungsi menghambat adhesi dari
monosit dan platelet, kemotaksis leukosit, dan oksidasi LDL. Tujuan: Membuktikan pengaruh pemberian ekstrak coklat sebagai antioksidan
terhadap regresi plak ateroma dinding arkus aorta tikus putih jantan (Rattus
norvegicus strain wistar) aterosklerotik
Metode: Eksperimental dengan menggunakan rancangan The Post Test Only
Control Group Design. Penelitian ini menggunakan 5 kelompok perlakuan yang
masing-masing terdiri dari 5 tikus (kontrol negatif, kontrol positif, ekstrak biji
coklat dosis 1,8mg/200gBB/hari, 3,6mg/200gBB/hari dan 5,4mg/200gBB/hari).
Preparat kemudian dilihat dan dianalisis ketebalan tunika intima dan media arkus
aortanya.
Hasil Penelitian: Uji Tukey 1% menujukkan bahwa pemberian dosis
3,6mg/200gBB/hari mempunyai efek yang sama dengan dosis
5,4mg/200gBB/hari. Pemberian ekstrak biji coklat berpengaruh sangat kuat secara
negatif terhadap ketebalan dinding aorta (R= -0,862). Analisis regresi linier
menunjuk dosis 6,28mg/200gBB/hari tikus sebagai dosis optimal pada penelitian
ini.
Kesimpulan: Ekstrak biji coklat (Theobroma cacao) terbukti mempunyai
aktivitas antioksidan yang dapat meregresi plak ateroma dinding arkus aorta tikus
putih jantan aterosklerotik.
Kata Kunci: Biji Coklat, Flavonoid, Nitric Oxide, Aterosklerosis.
* : Guru Besar Ilmu Penyakit Dalam konsultan Penyakit Tropis dan Infeksi, FK UMM
** : Staff pengajar Ilmu Penyakit Mata, FK UMM
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ABSTRACT
Muyasyarahma, Sidika Yunia. 2014. The Effect of Cocoa Extract (Theobroma
cacao) as An Antioxidant To Regress The Atheroma Plaque of Aortic Arc
Wall In Atherosclerotic Male White Rats (Rattus norvegicus strain wistar).
Final project, Medical Faculty of Muhammadiyah Malang University.
Advisors: (1) Djoni Djunaedi* (2) Yuliono Trika Nur Hasan **
Background: Coronary Heart Disease (CHD) has been proven as the main cause
of death in the world. The major cause of CHD is the narrowing of proximal
coronary artery caused by atherosclerosis may due to the decreasing of antioxidant
fraction that has been identified in cacao beans which stimulates the production of
Nitric Oxide (NO) which posses an ability to prevent leukocytes and platelet
adhesion, leukocyte chemotactic and Low Density Lipoprotein (LDL) oxidation.
Purpose: Proving the effect of cocoa extract as an antioxidant to regress the
atheroma plaque of aortic arc wall in atherosclerotic male white rats.
Method: The study was experimental with The Post Test Only Control Group
Design which consisted of 5 groups, each group consisted of 5 rat, (negative
control, positive control, and cocoa extract 1,8mg/200gBW/day,
3,6mg/200gBW/day and 5,4mg/200gBW/day). Tunica intima and media thickness
of the blood vessels were observed and analyzed.
Result: Tukey 1% test showed that dose 3,6mg/200gBB/day had the same effect
as dose 5,4mg/200gBB/day. Cocoa extract had very strong negative effect on the
arcus aortic wall thickness (R= -0,862). The linear regression analysis pointed to a
dose of 6,28mg/200gBB/day as the optimal dose in this study.
Conclusion: Cocoa extract approved have a potential antioxidant activity to
regress the atheroma plaque of aortic arc wall in atherosclerotic male white rats.
Key words: Cacao beans, Flavonoid, Nitric Oxide, Atherosclerosis
* : Professor of The Internal Medicine Department, Tropico-Infectiologist Consultant,
FK UMM
** : Lecturer of The Eye Medicine Department, FK UMM
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DAFTAR ISI
LEMBAR PENGESAHAN………………………………………………. i
LEMBAR PENGUJIAN………………………………………………….. ii
KATA PENGANTAR…………………………………………………….. iii
ABSTRAK……………………………………………………………….... v
ABSTRACT……………………………………………………………….... vi
DAFTAR ISI………………………………………………………………. vii
DAFTAR SINGKATAN.............................................................................. x
DAFTAR GAMBAR.................................................................................... xi
DAFTAR TABEL ........................................................................................ xii
BAB 1 PENDAHULUAN........................................................................... 1
1.1 Latar Belakang......................................................................................... 1
1.2 Rumusan Masalah.................................................................................... 3
1.3 Tujuan Penelitian..................................................................................... 3
1.3.1 Tujuan Umum................................................................................ 3
1.3.2 Tujuan Khusus............................................................................... 4
1.4 Manfaat Penelitian................................................................................... 4
1.4.1 Manfaat Akademik........................................................................ 4
1.4.2 Manfaat Klinis............................................................................... 4
BAB 2 TINJAUAN PUSTAKA..................................................................... 5
2.1 Aterosklerosis ............................................................................................ 5
2.1.1 Definisi ………………………………………………………..... 5
2.1.2 Etiologi ………………………………………………………… 6
2.1.3 Faktor Resiko …………………………………………………... 6
2.1.4 Klasifikasi Lesi Aterosklerosis ………………………………… 10
2.1.5 Sel-Sel yang Terlibat dalam Aterosklerosis ………………….... 12
2.1.6 Patogenesis Aterosklerosis …………………………………….. 14
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2.2 Nitrite Oxide (NO) .................................................................................. 17
2.2.1 Produksi NO ............................................................................... 18
2.2.2 Fungsi NO .................................................................................. 18
2.3 Reactive Oxygen Species (ROS) .............................................................. 19
2.4 Coklat (Theobroma cacao) ...................................................................... 21
2.4.1 Taksonomi .................................................................................. 21
2.4.2 Morfologi .................................................................................... 21
2.4.3 Biji Theobroma cacao ................................................................ 23
2.4.4 Manfaat Biji Theobroma cacao ................................................. 24
2.5 Peran Flavonoid Biji Theobroma cacao sebagai Antioksidan ................ 25
2.5.1 Flavanol ...................................................................................... 26
2.5.2 Procyanidin ................................................................................. 27
2.6 Bioavaibilitas Biji Coklat (Theobroma cacao) ........................................ 29
BAB 3 KERANGKA KONSEP DAN HIPOTESIS .................................... 30
3.1 Kerangka Konsep ...................................................................................... 30
3.2 Hipotesis ................................................................................................... 32
BAB 4 METODE PENELITIAN .................................................................. 33
4.1 Rancangan Penelitian ................................................................................ 33
4.2 Waktu dan Tempat Pelaksanaan Penelitian .............................................. 33
4.3 Populasi dan Sampel Penelitian ................................................................ 33
4.3.1 Populasi ......................................................................................... 33
4.3.2 Sampel ........................................................................................... 33
4.3.3 Teknik Sampling ........................................................................... 33
4.3.4 Estimasi Besar Sampel .................................................................. 34
4.4 Karakteristik sampel penelitin .................................................................. 35
4.5 Variabel dan Definisi Operasional ............................................................ 35
4.5.1 Variabel ........................................................................................ 35
4.5.2 Definisi Operasional ..................................................................... 35
4.6 Dasar penentuan dosis Ekstrak Biji Coklat .............................................. 36
4.7 Bahan dan Instrumen penelitian ................................................................ 37
4.7.1 Bahan ............................................................................................. 37
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4.7.2 Instrumen ....................................................................................... 38
4.8 Alur penelitian .......................................................................................... 40
4.9 Prosedur penelitian .................................................................................... 41
4.9.1 Pembagian Kelompok Tikus ......................................................... 41
4.9.2 Adaptasi ......................................................................................... 41
4.9.3 Pemberian Diet Aterogenik ........................................................... 41
4.9.4 Pemberian Ekstrak Biji Coklat ..................................................... 42
4.9.5 Proses Anastesi Dan Pembedahan Hewan Coba ........................... 42
4.9.6 Pembuatan Sediaan Histopatologi ................................................. 43
4.9.7 Analisa Data .................................................................................. 44
BAB 5 HASIL PENELITIAN DAN ANALISIS DATA.............................. 47
5.1 Gambaran Mikroskopis Dinding Arkus Aorta.............................. 47
5.2 Analisis Data ............................................................................... 49
BAB 6 PEMBAHASAN............................................................................ ..... 54
BAB 7 KESIMPULAN DAN SARAN .......................................................... 59
7.1 Kesimpulan ................................................................................... 59
7.2 Saran .............................................................................. ............... 59
DAFTAR PUSTAKA
LAMPIRAN
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DAFTAR SINGKATAN
Apo (a) : Apolipoprotein (a)
BH4 : Tetrahydrobiopterin
CAD : Coronary Artery Disease
EDRF : Endothelium Derived Relaxing Factor
EGF : Epidermal Growth Factor
ET : Endothelin-1
FGF : Fibroblast Growth Factor
HDL : High Density Lipoprotein
HoCys : Homocysteins
LDL : Low Density Lipoprotein
LDL-C : Low Density Lipoprotein-Cholesterol
M-CSF : Macrophage Colony Stimulating Factor
NO : Nitric Oxide
eNOS : endothelial Nitric Oxide Synthase
Ox-LDL : Oxidized LDL
PDGF : Platelet Derived Growth Factor
PGE2 : Prostaglandin E2
PGG2 : Prostaglandin G2
PGH2 : Prostaglandin H2
PGI2 : Prostaglandin I2
ROS : Reactive Oxygen Species
s-ICAM : soluble- Intracellular Adhesion Molecule
s-VCAM : soluble- Vascular Cellular Adhesion Molecule
TGF-β : Transforming Growth Factor-β
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DAFTAR GAMBAR
Gambar 2.1 Pembuluh Darah Aterosklerotik ........................................... 5
Gambar 2.2 Patofisiologi Aterosklerosis ................................................. 16
Gambar 2.3 Theobroma cacao ................................................................ 21
Gambar 2.4 Biji Theobroma cacao ......................................................... 23
Gambar 2.5 Komponen Flavonoid .......................................................... 25
Gambar 2.6 Struktur Kimia Flavanol ...................................................... 26
Gambar 2.7 Struktur procyanidin ............................................................ 27
Gambar 2.8 Mekanisme Antioksidan Biji Coklat ................................... 29
Gambar 5.1 Gambaran Mikroskopis Ketebalan Dinding Arkus Aorta
Tiap Perlakuan (Perbesaran 400x) ………………………… 47
Gambar 5.2 Kurva Hubungan Antar Dosis Ekstrak Biji Coklat dengan
Ketebalan Plak Tunika Intima dan Media Dinding Arkus
Aorta ……………………………………….……………... 51`
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DAFTAR TABEL
Tabel 4.1 Konversi Dosis Antar Spesies ..................................................... 36
Tabel 5.1 Pengaruh Ekstrak Biji Coklat terhadap Ketebalan Plak Tunika
Intima dan Media Dinding Arkus Aorta Setelah Perlakuan…… 48
Tabel 5.2 Perbandingan Dosis Ekstrak Biji Coklat terhadap Ketebalan Plak
Tunika Intima dan Media Dinding Arkus Aorta Setelah
Perlakuan …………………………………………………….... 50
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