PEMBERIAN EKSTRAK TEH (Camellia sinensis) HIJAU PER ORAL MENCEGAH PENINGKATAN EKSPRESI MMP-1 DAN PENURUNAN JUMLAH KOLAGEN LEBIH BANYAK DARIPADA EKSTRAK TEH OOLONG PADA MENCIT BALB-C (Mus musculus) YANG DIPAPAR SINAR UV-B.
TESIS
PEMBERIAN EKSTRAK TEH (Camellia sinensis) HIJAU PER ORAL
MENCEGAH PENINGKATAN EKSPRESI MMP-1 DAN PENURUNAN
JUMLAH KOLAGEN LEBIH BANYAK DARIPADA EKSTRAK TEH
OOLONG PADA MENCIT BALB-C (Mus musculus) YANG DIPAPAR
SINAR UV-B
HERMAWAN ADIGUNA
PROGRAM PASCASARJANA
UNIVERSITAS UDAYANA
DENPASAR
2016
LEMBAR PENGESAHAN
TESIS INI TELAH DISETUJUI
TANGGAL
Pembimbing I,
Pembimbing II,
Prof. dr. IGM Aman, Sp.FK
Dr. dr A.A.G.P.Wiraguna SpKK(K).,FINSD,FAADV
NIP:194606191976021001
NIP:195609121984121001
Mengetahui
Ketua Program Studi Ilmu Biomedik
Program Pascasarjana
Universitas Udayana,
Dr. dr. Gde Ngurah Indraguna Pinantih, Msc, Sp.GK
NIP: 195805211985031002
DAFTAR ISI
HALAMAN JUDUL……………………………………………………………....i
PRASYARAT GELAR……………………………………………………………ii
LEMBAR PENGESAHAN……………………………………………………….iii
LEMBAR PENETAPAN PENGUJI……………………………………………...iv
UCAPAN TERIMA KASIH……………………………………………………...v
ABSTRAK.............................................................................................................viii
DAFTAR ISI ............................................................................................................x
DAFTAR GAMBAR .................................................................................... .......xiv
DAFTAR TABEL...................................................................................................xv
DAFTAR SINGKATAN DAN GAMBAR……………………………………..xvi
DAFTAR LAMPIRAN ................................................................................ .......xvii
BAB I PENDAHULUAN ........................................................................................1
1.1 Latar Belakang................................................................................................1
1.2 Rumusan Masalah ..........................................................................................5
1.3 Tujuan Penelitian ............................................................................................5
1.3.1 Tujuan Umum…………………………………………........................5
1.3.2 Tujuan Khusus………………………………………………………...5
1.4 Manfaat Penelitian ..........................................................................................6
1.4.1 Manfaat Ilmiah………………………………………………………...6
x
1.4.1 Manfaat Aplikasi………………………………………………………6
BAB II KAJIAN PUSTAKA ...................................................................................7
2.1 Penuaan...........................................................................................................7
2.1.1 Faktor Penuaan ........................................................................................8
2.1.2 Teori Penuaan ..........................................................................................9
2.1.3 Gejala Klinis Penuaan............................................................................12
2.1.4 Pencegahan Penuaan ..............................................................................15
2.2 Radikal Bebas ...............................................................................................17
2.3 Sinar Ultraviolet ...........................................................................................18
2.3.1 Ultraviolet B ..........................................................................................20
2.4 Antioksidan...................................................................................................23
2.5 Kolagen dan MMP-1 .....................................................................................27
2.6 Teh Oolong Dan Teh Hijau ...........................................................................30
2.7 Pengaruh Teh Hijau Terhadap Kolagen .......................................................31
2.8 TIMP.............................................................................................................38
2.9 Mencit ...........................................................................................................38
BAB III KERANGKA BERPIKIR, KONSEP DAN HIPOTESIS PENELITIAN 40
3.1 Kerangka Berpikir ........................................................................................40
3.1.1 Kerangka Pikir Penelitian ......................................................................40
3.2 Konsep Penelitian .........................................................................................42
3.3 Hipotesis Penelitian ......................................................................................43
xi
BAB IV METODOLOGI PENELITIAN ..............................................................44
4.1 Rancangan Penelitian ...................................................................................44
4.2 Tempat Dan Waktu Penelitian......................................................................46
4.3 Populasi Dan Sampel ....................................................................................46
4.3.1 Populasi .................................................................................................46
4.3.2 Kriteria Sampel ......................................................................................47
4.3.2.1 Kriteria Inklusi ....................................................................................47
4.3.2.2 Kriteria Drop Out ................................................................................47
4.4 Besar Sampel………………………………………………………............47
4.5 Variabel Penelitian .......................................................................................48
4.5.1 Klasifikasi Variabel ...............................................................................48
4.5.2 Definisi Operasional Variabel ...............................................................49
4.6 Alat Bahan Dan Hewan Percobaan ..............................................................52
4.6.1 Alat Penelitian .......................................................................................52
4.6.2 Bahan Penelitian ....................................................................................52
4.6.3 Hewan Percobaan ..................................................................................52
4.7 Prosedur Penelitian .......................................................................................53
4.7.1 Persiapan Hewan Uji .............................................................................53
4.7.2 Pembuatan Sediaan Histologis ..............................................................55
4.7.3 Pewarnaan dengan Sirius Red ...............................................................56
4.7.4 Pengamatan Hasil ..................................................................................57
4.7.5 Prosedur Penghitungan Ekspresi Kolagen Dermis ................................57
xii
4.7.6 Pengecatan Imunohistokimia MMP-1 ...................................................58
4.8 Alur Penelitian ..............................................................................................60
4.9 Analisis Data ................................................................................................61
BAB V HASIL PENELITIAN ..............................................................................62
5.1 Analisis Deskriptif ........................................................................................63
5.2 Uji Normalitas Data ......................................................................................66
5.3 Uji Homogenitas Data ..................................................................................67
5.4 Uji Komparibilitas ........................................................................................67
BAB VI PEMBAHASAN ......................................................................................69
6.1 Subjek Penelitian ..........................................................................................69
6.2 Distribusi dan Homogenitas Data Hasil Penelitian ......................................70
6.3 Pengaruh Pemberian Ekstrak Teh Hijau dan Teh Oolong Terhadap Jumlah
Kolagen Kulit................................................................................................70
6.4 Pengaruh Pemberian Ekstrak Teh Hijau dan Teh Oolong Terhadap Ekspresi
MMP-1..........................................................................................................75
BAB VII SIMPULAN DAN SARAN ...................................................................80
7.1 Simpulan .......................................................................................................80
7.2 Saran .............................................................................................................80
DAFTAR PUSTAKA ............................................................................................82
xiii
DAFTAR GAMBAR
Gambar 2.1 Efek akut dan kronis UVB.................................................................23
Gambar 2.2 Efek Radiasi UV pada Keratinosit (KC) dan Fibroblast (FB)...........25
Gambar 2.3 Reaksi Penghambatan antioksidan primer terhadap radikal lipida....27
Gambar 2.4 Antioksidan bertindak sebagai prooksidan pada konsentrasi tinggi..27
Gambar 2.5 Tiga Macam Cara Kerja Dari Antioksidan Alami.............................28
Gambar 2.6 Kolagen tipe I dengan pewarnaan
HE....................................................................................................30
Gambar 2.7 Cincin Aromatik................................................................................32
Gambar 3 Konsep Penelitian.................................................................................42
Gambar 4 Skema Rancangan Penelitian................................................................44
Gambar 5.1 Histopatologi Dermis Kelompok Perlakuan I………………………64
Gambar 5.2 Histopatologi Dermis Kelompok Perlakuan II……………………...65
Gambar 5.3 Ekspresi MMP-1 Jaringan Dermis Mencit Perlakuan 1(P1) dengan
pengecatan Imunohistokimia…...........................................................65
Gambar 5.4 Ekspresi MMP-1 Jaringan Dermis Mencit Perlakuan 1(P2) dengan
pengecatan Imunohistokimia…...........................................................66
xiv
DAFTAR TABEL
Table 5.1 Hasil Analisis Deskriptif Data Jumlah Kolagen dan Ekspresi MMP1……………………………………………………………………63
Tabel 5.2 Hasil Uji Normalitas Data Antar Kelompok………………………66
Tabel 5.3 Hasil Uji Homogenitas Data Antar Kelompok…………………….67
Tabel 5.4 Rerata Jumlah Kolagen Dan Ekspresi MMP-1 Antar
Kelompok.........................................................................................68
xv
DAFTAR SINGKATAN DAN LAMBANG
AAM
: Anti Aging Medicine
AP-1
: Activator Protein -1
ECM
: Extracellular Matrix
FB
: Fibroblas
KC
: Keratinosit
mJ/Cm2
: mili joule per sentimeter persegi
MMP
: Matrix Metalloproteinase -1
ROS
: Reactive Oxygen Species
SOD
: Superoxyde Dismutase
TGF-β
: Tumor Growth Factor- β
UVB
: Ultra Violet B
xvi
DAFTAR LAMPIRAN
Lampiran 1. Keterangan Kelayakan Etik ...................................................................87
Lampiran 2. Laporan Hasil Analisis Fitokimia Teh Hijau dan Teh Oolong…....... ..88
Lampiran 3. Analisis Statistik Kolagen Teh Hijau dan Teh Oolong ........................... ..89
Lampiran 4. Analisis Statistik MMP-1 Teh Hijau dan Teh Oolong……………..….92
Lampiran 5. Foto Aktivitas Penelitian .................................................................... ...96
xvii
PEMBERIAN EKSTRAK TEH (Camellia sinensis) HIJAU PER ORAL
MENCEGAH PENINGKATAN EKSPRESI MMP-1 DAN PENURUNAN
JUMLAH KOLAGEN LEBIH BANYAK DARIPADA EKSTRAK TEH
OOLONG PADA MENCIT BALB-C (Mus musculus) YANG DIPAPAR
SINAR UV-B
HERMAWAN ADIGUNA
PROGRAM PASCASARJANA
UNIVERSITAS UDAYANA
DENPASAR
2016
LEMBAR PENGESAHAN
TESIS INI TELAH DISETUJUI
TANGGAL
Pembimbing I,
Pembimbing II,
Prof. dr. IGM Aman, Sp.FK
Dr. dr A.A.G.P.Wiraguna SpKK(K).,FINSD,FAADV
NIP:194606191976021001
NIP:195609121984121001
Mengetahui
Ketua Program Studi Ilmu Biomedik
Program Pascasarjana
Universitas Udayana,
Dr. dr. Gde Ngurah Indraguna Pinantih, Msc, Sp.GK
NIP: 195805211985031002
DAFTAR ISI
HALAMAN JUDUL……………………………………………………………....i
PRASYARAT GELAR……………………………………………………………ii
LEMBAR PENGESAHAN……………………………………………………….iii
LEMBAR PENETAPAN PENGUJI……………………………………………...iv
UCAPAN TERIMA KASIH……………………………………………………...v
ABSTRAK.............................................................................................................viii
DAFTAR ISI ............................................................................................................x
DAFTAR GAMBAR .................................................................................... .......xiv
DAFTAR TABEL...................................................................................................xv
DAFTAR SINGKATAN DAN GAMBAR……………………………………..xvi
DAFTAR LAMPIRAN ................................................................................ .......xvii
BAB I PENDAHULUAN ........................................................................................1
1.1 Latar Belakang................................................................................................1
1.2 Rumusan Masalah ..........................................................................................5
1.3 Tujuan Penelitian ............................................................................................5
1.3.1 Tujuan Umum…………………………………………........................5
1.3.2 Tujuan Khusus………………………………………………………...5
1.4 Manfaat Penelitian ..........................................................................................6
1.4.1 Manfaat Ilmiah………………………………………………………...6
x
1.4.1 Manfaat Aplikasi………………………………………………………6
BAB II KAJIAN PUSTAKA ...................................................................................7
2.1 Penuaan...........................................................................................................7
2.1.1 Faktor Penuaan ........................................................................................8
2.1.2 Teori Penuaan ..........................................................................................9
2.1.3 Gejala Klinis Penuaan............................................................................12
2.1.4 Pencegahan Penuaan ..............................................................................15
2.2 Radikal Bebas ...............................................................................................17
2.3 Sinar Ultraviolet ...........................................................................................18
2.3.1 Ultraviolet B ..........................................................................................20
2.4 Antioksidan...................................................................................................23
2.5 Kolagen dan MMP-1 .....................................................................................27
2.6 Teh Oolong Dan Teh Hijau ...........................................................................30
2.7 Pengaruh Teh Hijau Terhadap Kolagen .......................................................31
2.8 TIMP.............................................................................................................38
2.9 Mencit ...........................................................................................................38
BAB III KERANGKA BERPIKIR, KONSEP DAN HIPOTESIS PENELITIAN 40
3.1 Kerangka Berpikir ........................................................................................40
3.1.1 Kerangka Pikir Penelitian ......................................................................40
3.2 Konsep Penelitian .........................................................................................42
3.3 Hipotesis Penelitian ......................................................................................43
xi
BAB IV METODOLOGI PENELITIAN ..............................................................44
4.1 Rancangan Penelitian ...................................................................................44
4.2 Tempat Dan Waktu Penelitian......................................................................46
4.3 Populasi Dan Sampel ....................................................................................46
4.3.1 Populasi .................................................................................................46
4.3.2 Kriteria Sampel ......................................................................................47
4.3.2.1 Kriteria Inklusi ....................................................................................47
4.3.2.2 Kriteria Drop Out ................................................................................47
4.4 Besar Sampel………………………………………………………............47
4.5 Variabel Penelitian .......................................................................................48
4.5.1 Klasifikasi Variabel ...............................................................................48
4.5.2 Definisi Operasional Variabel ...............................................................49
4.6 Alat Bahan Dan Hewan Percobaan ..............................................................52
4.6.1 Alat Penelitian .......................................................................................52
4.6.2 Bahan Penelitian ....................................................................................52
4.6.3 Hewan Percobaan ..................................................................................52
4.7 Prosedur Penelitian .......................................................................................53
4.7.1 Persiapan Hewan Uji .............................................................................53
4.7.2 Pembuatan Sediaan Histologis ..............................................................55
4.7.3 Pewarnaan dengan Sirius Red ...............................................................56
4.7.4 Pengamatan Hasil ..................................................................................57
4.7.5 Prosedur Penghitungan Ekspresi Kolagen Dermis ................................57
xii
4.7.6 Pengecatan Imunohistokimia MMP-1 ...................................................58
4.8 Alur Penelitian ..............................................................................................60
4.9 Analisis Data ................................................................................................61
BAB V HASIL PENELITIAN ..............................................................................62
5.1 Analisis Deskriptif ........................................................................................63
5.2 Uji Normalitas Data ......................................................................................66
5.3 Uji Homogenitas Data ..................................................................................67
5.4 Uji Komparibilitas ........................................................................................67
BAB VI PEMBAHASAN ......................................................................................69
6.1 Subjek Penelitian ..........................................................................................69
6.2 Distribusi dan Homogenitas Data Hasil Penelitian ......................................70
6.3 Pengaruh Pemberian Ekstrak Teh Hijau dan Teh Oolong Terhadap Jumlah
Kolagen Kulit................................................................................................70
6.4 Pengaruh Pemberian Ekstrak Teh Hijau dan Teh Oolong Terhadap Ekspresi
MMP-1..........................................................................................................75
BAB VII SIMPULAN DAN SARAN ...................................................................80
7.1 Simpulan .......................................................................................................80
7.2 Saran .............................................................................................................80
DAFTAR PUSTAKA ............................................................................................82
xiii
DAFTAR GAMBAR
Gambar 2.1 Efek akut dan kronis UVB.................................................................23
Gambar 2.2 Efek Radiasi UV pada Keratinosit (KC) dan Fibroblast (FB)...........25
Gambar 2.3 Reaksi Penghambatan antioksidan primer terhadap radikal lipida....27
Gambar 2.4 Antioksidan bertindak sebagai prooksidan pada konsentrasi tinggi..27
Gambar 2.5 Tiga Macam Cara Kerja Dari Antioksidan Alami.............................28
Gambar 2.6 Kolagen tipe I dengan pewarnaan
HE....................................................................................................30
Gambar 2.7 Cincin Aromatik................................................................................32
Gambar 3 Konsep Penelitian.................................................................................42
Gambar 4 Skema Rancangan Penelitian................................................................44
Gambar 5.1 Histopatologi Dermis Kelompok Perlakuan I………………………64
Gambar 5.2 Histopatologi Dermis Kelompok Perlakuan II……………………...65
Gambar 5.3 Ekspresi MMP-1 Jaringan Dermis Mencit Perlakuan 1(P1) dengan
pengecatan Imunohistokimia…...........................................................65
Gambar 5.4 Ekspresi MMP-1 Jaringan Dermis Mencit Perlakuan 1(P2) dengan
pengecatan Imunohistokimia…...........................................................66
xiv
DAFTAR TABEL
Table 5.1 Hasil Analisis Deskriptif Data Jumlah Kolagen dan Ekspresi MMP1……………………………………………………………………63
Tabel 5.2 Hasil Uji Normalitas Data Antar Kelompok………………………66
Tabel 5.3 Hasil Uji Homogenitas Data Antar Kelompok…………………….67
Tabel 5.4 Rerata Jumlah Kolagen Dan Ekspresi MMP-1 Antar
Kelompok.........................................................................................68
xv
DAFTAR SINGKATAN DAN LAMBANG
AAM
: Anti Aging Medicine
AP-1
: Activator Protein -1
ECM
: Extracellular Matrix
FB
: Fibroblas
KC
: Keratinosit
mJ/Cm2
: mili joule per sentimeter persegi
MMP
: Matrix Metalloproteinase -1
ROS
: Reactive Oxygen Species
SOD
: Superoxyde Dismutase
TGF-β
: Tumor Growth Factor- β
UVB
: Ultra Violet B
xvi
DAFTAR LAMPIRAN
Lampiran 1. Keterangan Kelayakan Etik ...................................................................87
Lampiran 2. Laporan Hasil Analisis Fitokimia Teh Hijau dan Teh Oolong…....... ..88
Lampiran 3. Analisis Statistik Kolagen Teh Hijau dan Teh Oolong ........................... ..89
Lampiran 4. Analisis Statistik MMP-1 Teh Hijau dan Teh Oolong……………..….92
Lampiran 5. Foto Aktivitas Penelitian .................................................................... ...96
xvii