HASIL PENELITIAN

PENGARUH EKSTRAK JINTAN HITAM (Nigella Sativa) TOPIKAL

TETES MATA TERHADAP PENURUNAN EKSPRESI Nuclear Factor

Kappa Beta (NF-Kβ) PADA KORNEA TIKUS JANTAN (Rattus Novergicus

Strain Wistar) MODEL INFLAMASI

Oleh:

IMANIYAH HUSNI

201110330311151

UNIVERSITAS MUHAMMADIYAH MALANG

FAKULTAS KEDOKTERAN

2015

i

ii

KATA PENGANTAR

Puji syukur kehadirat Allah SWT atas rahmat dan inayah-Nya sehingga

penulis diberi kekuatan dan kemampuan dalam menyelesaikan karya tulis akhir

ini yang berjudul “PENGARUH EKSTRAK JINTAN HITAM (Nigella Sativa)

TOPIKAL TETES MATA TERHADAP PENURUNAN EKSPRESI Nuclear

Factor Kappa Beta (NF-Kβ) PADA KORNEA TIKUS JANTAN (Rattus

Novergicus Strain Wistar) MODEL INFLAMASI”. Karya tulis akhir ini diajukan

untuk memenuhi persyaratan Pendidikan Sarjana Fakultas Kedokteran Universitas

Muhammadiyah Malang.

Pada kesempatan ini, penulis ingin menyampaikan terimakasih kepada :

1. dr. Irma Suswati, M.Kes beserta para pembantu dekan selaku Dewan

Dekanat Fakultas Kedokteran UMM.

2. dr. Bragastio Sidharta, Sp.M, selaku pembimbing 1 atas kesabaran,

kebaikan hati serta kesediaan dalam meluangkan waktu dalam membantu,

membimbing dan mengarahkan penulis dalam menyelesaikan karya tulis

akhir ini.

3. dr. Thontowi Djauhari, NS, M.Kes, selaku pembimbing 2 atas kesabaran,

kebaikan hati serta kesediaan dalam meluangkan waktu dalam membimbing

penulis hingga dapat menyelesaikan menyelesaikan dengan baik.

4. dr. Heru Priyo Husodo, Sp.Rad, selaku penguji tugas akhir ini yang telah

membantu dan memberikan masukan kepada penulis dalam menyelesaikan

karya tulis akhir ini.

5. Seluruh dosen beserta karyawan-karyawati Fakultas Kedokteran UMM yang

selalu membantu dan membimbing penulis.

iii

6. Kedua orang tua penulis, Ayahanda Husni Husain Hasan dan Ibunda

Djasna Ali atas doa dan dukungannya yang tak pernah putus. Kedua saudari

penulis Inayati Husni dan Amaliyah Husni atas bantuan doa, motivasi dan

tenaganya beserta seluruh keluarga tercinta yang senantiasa setia

mendukung penulis dalam menyelesaikan karya tulis akhir ini.

7. Seluruh staf TU dan laboran, Mas Miftah dan Pak Joko yang bersedia

membantu setiap tahapan proses penelitian hingga penyelesaian tugas akhir

ini.

8. Sejawat FK UMM angkatan 2011 “Adorables” atas dukungan, bantuan, dan

kerjasamanya. Semoga kelak dapat menjadi dokter-dokter profesional yang

dapat bermanfaat bagi orang lain serta dapat menjalankan amanah sebagai

dokter karena Allah SWT.

9. Sahabat-sahabat terbaik yang selalu membantu selama penyelesaian karya

tulis akhir ini, Mariyah,Lilly, Tari, Alfi, Laksita, Putrinda, Kinanthi, Jeanny,

Ilma, Dwi, Friska, Hakimah, terimakasih atas doa, dukungan dan bantuan

yang telah diberikan selama ini.

10. Kelompok Jintan Hitam Lilly, Muqsit, Rofwiun atas dukungan dan

kerjasama dalam penyelesaian karya tulis akhir ini.

11. Keluarga besar Skill laboratorium FK UMM atas doa dan dukungannya.

12. Keluarga Besar Al-Zahrawi FK UMM atas doa, dukungan serta bantuan

yang diberikan selama ini.

13. Semua pihak yang terlibat secara langsung maupun tidak langsung,

terimakasih atas bantuan dan dukungannya.

iv

Penulis menyadari bahwa tugas akhir ini masih jauh dari kesempurnaan.

Oleh karena itu, penulis sangat mengharapkan saran, kritik dan masukan yang

membangun. Dengan mengharapkan Keridhaan-Nya, semoga karya tulis ini dapat

menambah wawasan keilmuan dan bermanfaat bagi semua pihak.

Malang, Agustus 2015

Penulis

v

DAFTAR ISI

Halaman

LEMBAR PENGESEHAN .....................................................................................i

LEMBAR PENGUJIAN .........................................................................................ii

KATA PENGANTAR ............................................................................................iii

ABSTRAK ..............................................................................................................vi

DAFTAR ISI ........................................................................................................viii

DAFTAR GAMBAR .............................................................................................xi

DAFTAR TABEL .................................................................................................xii

DAFTAR SINGKATAN ......................................................................................xiii

DAFTAR LAMPIRNAN ......................................................................................xiv

BAB 1 PENDAHULUAN

1.1 Latar Belakang .................................................................................................1

1.2 Rumusan Masalah .............................................................................................4

1.3 Tujuan Penelitian ..............................................................................................4

1.3.1 Tujuan Umum .........................................................................................4

1.3.2 Tujuan Khusus ........................................................................................4

1.4 Manfaat Penelitian ............................................................................................5

1.4.1 Akademis .................................................................................................5

1.4.2 Klinis .......................................................................................................5

BAB 2 TINJAUAN PUSTAKA

2.1 Kornea ...............................................................................................................6

2.1.1 Definisi ...................................................................................................6

2.1.2 Anatomi Kornea ......................................................................................7

2.2 Inflamasi .........................................................................................................10

2.2.1 Definisi ..................................................................................................10

2.2.2 Proses Inflamasi ....................................................................................11

2.3 Inflamasi Kornea ............................................................................................13

2.4 Jintan Hitam (Nigella sativa) ..........................................................................14

2.4.1 Definisi dan Taksonomi .........................................................................14

2.4.2 Komposisi Kimia Nigella sativa ...........................................................15

2.5 Thymoquinon ..................................................................................................17

viii

2.5.1 Definisi .................................................................................................17

2.5.2 Mekanisme Thymoquinon menghambat aktifasi

Nuclear Factor Kappa Beta (NF-Kβ) ...................................................18

2.6 Nuclear Factor Kappa Beta (NF-Kβ) ............................................................19

2.6.1 Definisi ..................................................................................................19

2.6.2 Struktur Nuclear Factor Kappa Beta (NF-Kβ) .....................................21

2.6.3 Mekanisme Nuclear Factor Kappa Beta (NF-Kβ) ...............................22

2.6.5 Aktivasi Nuclear Factor Kappa Beta (NF-Kβ) ....................................23

2.6.6 Jalur Aktivasi Nuclear Factor Kappa Beta (NF-Kβ) ............................24

BAB 3 KERANGKA KONSEP DAN HIPOTESIS

3.1 Kerangka Konsep ............................................................................................27

3.2 Hipotesis .........................................................................................................28

BAB 4 METODOLOGI PENELITIAN

4.1 Rancangan Penelitian .....................................................................................29

4.2 Tempat dan Waktu Penelitian ........................................................................29

4.3 Populasi dan Sampel .......................................................................................29

4.3.1 Populasi .................................................................................................29

4.3.2 Sampel ...................................................................................................29

4.3.3 Besar Sampel Penelitian .......................................................................30

4.3.4 Teknik Pengambilan Sampel ................................................................32

4.3.5 Karakteristik Sampel Penelitian ............................................................32

4.3.6 Variabel Penelitian ................................................................................33

4.3.7 Definisi Operasional Variabel Penelitian ..............................................33

4.4 Alat dan Bahan ...............................................................................................34

4.3.1 Alat .......................................................................................................34

4.3.2 Bahan ...................................................................................................34

4.5 Kerangka Operasional Penelitian ...................................................................35

4.6 Prosedur Penelitian ........................................................................................36

4.6.1 Adaptasi ................................................................................................36

4.6.2 Pembuatan Ekstrak Nigella sativa ........................................................36

4.6.3 Pembuatan Solusio Nigella sativa Tetes Mata Topikal .........................37

4.6.4 Perlakuan Sampel ..................................................................................37

ix

4.6.5 Pengambilan Sampel Kornea untuk Pengamatan

Ekspresi NF-Kβ .....................................................................................38

4.6.6 Pemeriksaan NF-Kβ dalam Metode Imunohistokimia .........................39

4.6.7 Pengamatan Ekpresi NF-Kβ dengan

Metode Imunhistokimia pada Kornea ...................................................40

4.6.8 Penghitungan Ekspresi NF-Kβ ..............................................................41

4.7 Analisa Data ...................................................................................................42

BAB 5 HASIL PENELITIAN DAN ANALISIS DATA

5.1 Hasil Penelitian ...............................................................................................37

5.2 Analisis Data ....................................................................................................38

5.2.1 Uji Kruskal Wallis ..................................................................................38

5.2.2 Uji Post-Hoc (Mann-Whitney) ...............................................................39

5.2.3 Uji Korelasi …………..........................................................................39

BAB 6 PEMBAHASAN ........................................................................................41

BAB 7 KESIMPULAN DAN SARAN

7.1 Kesimpulan ......................................................................................................52

7.2 Saran ................................................................................................................52

DAFTAR PUSTAKA ............................................................................................53

LAMPIRAN ...................... ...................................................................................59

x

DAFTAR GAMBAR

Gambar Halaman

2.1 Anatomi Mata ...................................................................................................7

2.2 Anatomi Kornea ..............................................................................................9

2.3 Jintan Hitam (a) bunga dan (b) biji .................................................................14

2.4 Struktur Kimia Thymoquinon .........................................................................17

2.5 Mekanisme Thymoquinon Menghambat Aktifasi

Nuclear Factor Kappa Beta (NF-Kβ) ...........................................................18

2.6 Protein Nuclear Factor Kappa Beta (NF-Kβ) ................................................20

2.7 Kelas Nuclear Factor Kappa Beta (NF-Kβ) .................................................21

2.8 Mekanisme Nuclear Factor Kappa Beta (NF-Kβ) .........................................22

2.9 Jalur Aktifasi Nuclear Factor Kappa Beta (NF-Kβ) ......................................25

3.1 Kerangka Konsep ...........................................................................................26

4.1 Kerangka Operasional Penelitian ...................................................................34

4.2 Lokasi Pengambilan Sampel Kornea ..............................................................38

5.1 Gambaran ekspresi NF-Kβ ............................................................................44

5.2 Ekspresi NF-Kβ ..............................................................................................45

xi

DAFTAR TABEL

Tabel Halaman

2.1 Komposisi Kimia Nigella sativa .....................................................................16

2.2 Komposisi Minyak Nigella sativa ...................................................................16

4.1 Klasifikasi perhitungan NF-Kβ ........................................................................40

4.2 Jadwal Penelitian ............................................................................................41

5.1 Hasil Penelitian Ekspresi NF-Kβ pada Kornea

Mata Tikus (Rattus novergius strain wistar) Jantan .......................................44

5.2 Hasil Kruskal Wallis .......................................................................................46

5.3 Hasil Uji Post-Hoc (Mann-Whitney) ...............................................................46

xii

DAFTAR SINGKATAN

a-MSH : Alpha-Melanosit Stimulating Hormon

BAFF : B Cell Activating Factor

DBD : DNA-Binding Domain

DNA : Deoxyribonucleic Acid

DTQ : Dimmer Thymoquinone

IKK : IKB Kinase

NF-Kβ : Nuclear Factor Kappa Betta

NIK : NFkβ-Inducing Kinase

NLS : Localization Signals

NSO : Nigella sativa Oil

TGF : Transformidezzing beta Growth Factor

THQ : Thymohydroquinone

THY : Timol

TLR : Toll-Like Receptor

TNF : Tumor Necrosis Factor

TNFR : Tumor Necrosis Factor Receptor

TQ : Thymoquinone

RANK : Receptor Activator of Nuclear Factor Kb

RE : Response Elements

RHD : Rel Homology Domain

xiii

DAFTAR LAMPIRAN

Halaman

Lampiran 1 : Data Hasil Penelitian ......................................................................59

Lampiran 2 : Analisis Data ..................................................................................60

Lampiran 3 : Dokumentasi Kegiatan ....................................................................66

Lampiran 4 : Surat Determinasi Tanaman ............................................................70

Lampiran 5 : Surat Keterangan Ekstrak Jintan Hitam .........................................71

Lampiran 6 : Surat Keterangan Laboratorium Biomedik .....................................72

Lampiran 7 : Surat Keterangan Kelaikan Etik ......................................................73

Lampiran 8 : Surat Keterangan Laboratorium Biomedik UB ...............................74

Lampiran 9 : Surat Keterangan Pembacaan PA ....................................................75

Lampiran 10 : Kartu Konsultasi Tugas Akhir .......................................................76

xiv

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