BAB 5 KESIMPULAN DAN SARAN
5.1 Kesimpulan
1. Ekstrak etanol jinten hitam dosis 125mgkgBB, 250 mgkgBB dan 500 mg kgBB mampu mempengaruhi jumlah total leukosit dan
mempengaruhi persentase limfosit 2. Ekstrak etanol jinten hitam dosis 125mgkgBB, 250 mgkgBB dan 500
mg kgBB tidak menunjukkan pengaruh terhadap persentase monosit dan kadar IL-
1β 3. Ekstrak etanol jinten hitam dosis 125mgkgBB, 250 mgkgBB dan 500
mg kgBB mampu mempengaruhi jumlah total leukosit dan persentase limfosit
5.2 Saran
Dilakukan uji lanjutan untuk melihat lama penurunan jumlah total leukosit setelah pemberian ekstrak etanol jinten hitam.
40
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DAFTAR PUSTAKA
Abbas, Abul K, dan Andrew H. Lichtman, 2006. Basic Immunology 2nd Edition. Elsevier – Health Sciences Div
Abdulelah, H.A.A. dan Zainal-Abidin, B. A. H. 2007. In Vivo Anti-malarial Tests of Nigella sativa Black Seed Different Extracts
. Science Publications. 2 2: 46- 50
Adamu, harami, Ekanem, Bulama. 2010. Identification of Essential Oil Components from Nigella sativa Seed by Gas Chromatography-Mass
Spectroscopy . Abubakar Tafawa Balewa University. 9 10: 966-967.
Aggarwal, Bharat B., Kunnumakkara, Ajaikumar B. 2009. Molecular Targets and Therapeutic Uses of Spices Modern Uses for Ancient Medicine
. World Scientific. Hal. 258-260.
Anington LR. 1972. Introductory Laboratory Animal Science. The Breeding, Care and Managenment of Experinmenta1 Animals
. New York: The Interstate Printers Publishers Inc.
Arifiani AA. 2012. Karakterisasi Simplisia Dan Standardisasi Ekstrak Etanol Biji
Jinten Hitam Nigella sativa L.. Uin Syarif Hidayatullah Jakarta.
Arora, rajesh. 2008. Herbal Radiomodulators Applications in Medicine, Homeland Defence and Space.
CAB International. Hal. 76-77. Aslam M, Khan M, Ahmad S. 2011. In Vitro Bactericidal Activity Of Seeds
Extract Of Nigella sativa Against Methicillin Resistant Staphylococcus aureus Isolated From Tertiary Care Hospital Delhi Ncr Region India.
IJRPD 90-93 Anandika, D.W. 2011. Ekstrak Bawang Putih Allium sativum Menurunkan
Jumlah Leukosit pada Mencit Model Sepsis akibat Paparan Staphylococcus aureus
. CDK 183.Vol.38:2
41
UIN Syarif Hidayatullah Jakarta
Anderson SP, Lorraine MW. 2006. Patofisiologi Konsep Klinis Proses-Proses Penyakit Edisi 6.
Jakarta. EGC. Azis S. 2011. Standardisasi Bahan Obat Alam. Yogyakarta : Graha Ilmu
Aziz AEA, El Sayed NS, Mahran LG. 2011. Anti-Asthmatic And Anti-Allergic Effects Of Thymoquinone On Airway-Induced Hypersensitivity In Experimental
Animals. Journal of Applied Pharmaceutical Science 01 08; 2011: 109-117. Banacerrfa B, Unanue . 1979. Textbook of Immunology
Bloom, Fawcet. 1994. A Textbook of Histology. Penerjemah: Jan Tambayong. Jakarta: Penerbit Buku Kedokteran EGC . 409
Boraschi, D., Villa, L., Volpini, G., BossaÁ, P., Censini, S., Ghiara, P., Scapigliat, G., Nencioni, L., Bartalini, M., and Matteucci, G. 1990. Differential activity Of
Interleukin 1 Alpha And Interleukin 1 Beta In The Stimulation Of The Immune Response In Vivo
. Eur J Immunol 20, 317±321. Buriro, M.A., Tayyab, M., Ditta, A. Effect Of Nigella sativa On Serum
Cholesterol Of Albino Rat . Professional Med J Mar 2011;181: 142-146
Crowther JR. 2001. The ELISA Guidebook. New Jersey : Humana Press. Cruse JM, Lewis RE. 2003. Atlas Of Immunology. Florida: CRC Press
Departemen Kesehatan RI. 1995. Farmakope Indonesia Edisi IV. Direktorat Jendral Pengawasan Obat dan Makanan : Jakarta
Departemen Kesehatan RI. 2008. Farmakope Herbal Indonesia Edisi I. Direktorat Jendral Pengawasan Obat dan Makanan : Jakarta. Hal. 174-177.
Departemen Kesehatan RI. 1979. Materia Medika Indonesia Jilid III. Direktorat Jendral Pengawasan Obat dan Makanan : Jakarta. Hal. 114-117.
Departemen Kesehatan RI. 1985. Tanaman Obat Indonesia Jilid I. Direktorat Jendral Pengawasan Obat dan Makanan : Jakarta. Hal. 33.
UIN Syarif Hidayatullah Jakarta
Departemen Kesehatan RI. 2000. Parameter Standar Umum Ekstrak Tumbuhan Obat
. Direktorat Jendral Pengawasan Obat dan Makanan : Jakarta. Departemen Kesehatan RI. 1989. Vadmekum Bahan Obat Alam. Direktorat
Jendral Pengawasan Obat dan Makanan : Jakarta. Hal. 99-100. Dinarello CA. IL-
β. Department of Infectious Diseases, University of Colorado Health Sciences Center.
Donatus IA. 1983, Peranan Farmakologi Dalam Pengembangan Obat Tradisional
, oleh Husin, M, Risalah Simposium Penelitian Tumbuhan Obat III, Fakultas Farmasi Gajah Mada, Jogjakarta.
El Bagir. N. 2010. Immune Response and Pasteurella Resistance in Rabbits Fed Diets Containing Various Amounts of Black Cumin Seeds
. Department of Biochemistry, Faculty of Veterinary Medicine. 5 2: 163-167.
Farnsworth, R. Norman. 1966. Biological and Phytochemical Screening of Plants. Wiley Company
Ganong WF. 2003. Buku Ajar Fisiologi Kedokteran. Ed ke-20. Widjajakusumah HMD et al. penerjemah; Widjajakusumah HMD, editor. Jakarta: EGC.
Terjemahan dari: Review of Medical Physiology. Gerige, Saptha., Gerige, Mahesh., 2008. GC-MS Analysis of Nigella sativa Seeds
and Antimicrobial Activity of its Volatile oil. Departament of Biotecnology; Sri
Krishnadeveraya University; Anantapura - A. P. – India. pp. 1189-1192. Ghonime M, Eldomany R, Abdelaziz A, Soliman H, 2011. Evaluation Of
Immunomodulatory Effect Of Three Herbal Plants Growing In Egypt . Department
of Microbiology and Immunology, Helwan University: Helwan. 331:141 – 145. Gilani Hassan A. 2004. A Review Of Medicinal Uses and Pharmacological
Activities of Nigella sativa L. Departement of Biological and Biomedical Science.
7 4: 441-451.
UIN Syarif Hidayatullah Jakarta
Gartner LP, Hiatt JL, Strum JM, 2012.Essential Biologi Sel dan Histologi. Penerjemah : Fajar Arifin Gunawijaya. Pamulang: Bina Rupa Aksara Publisher
Hailat N., Al-Kahil S., Alkofahi A., Lafi S., Al-Ani F., Al-Darraji A., Bataineh Z. 1998. Effects of Nigella sativa Extracts on Antibody Response of Rats Vaccinated
with Brucella Vaccine. Faculty of Pharmacy Jordan University. pp. 217-221.
Halawani, Eman. 2009. Antibacterial Activity of Thymoquinone and Thymohydroquinone
of Nigella sativa L. and Their Interaction with Some Antibiotics.
Department of Biology Faculty of Science Taif University. 3 5-6: 148-152.
Haq A, Lobo PI, Al Tufail M, Rama NR and Al-Sedairy ST. Immunmodulatory effect of Nigella sativa proteins fractionated by ion exchange chromatography. Int
J Immunopharmacology 1999; 21: 288-5 Haq A, Abdullah M, Lobo PI, Al Tufail M, Khalid SA, Khabar, Kirtikant V.
Sheth, and Al-Sedairy ST. Nigella sativa: effect on human lymphocytes and polymorphonuclear leukocyte phagocytic activity. Int J Immunopharmacology
301995: 147-155. Harborne, J.B. 1987. Metode Fitokimia Penuntun Cara Modern Menganalisis
Tumbuhan . Penerbit ITB : Bandung. Hal 6-9.
Helmi, A., Anggraini, N., Handayani, D., Rasyid, R., 2006, Standarisasi Ekstrak Etanol Daun Eugenia cumini Merr.,J. Sains Tek. Far., 112
Isselbacher, dkk. Harrison: Prinsip-Prinsip Ilmu Penyakit Dalam. Jakarta: EGC;1999:2
Junquira Carlos, Carneiro Jose, Kelly Robert O. 1998. Basic Histology. Penerjemah: Jan Tambayong. Jakarta: Penerbit Buku Kedokteran EGC. 270.
Johnson GA, Zielger RJ, Hawley L. 2011. Essential Mikrobiologi dan Imunologi. Tangerang Selatan: Binarupa Aksara Publisher
UIN Syarif Hidayatullah Jakarta
Kanter, Mehmet. dkk. 2005. Gastroprotective Activity Of Nigella sativa L Oil And
Its Constituent, Thymoquinone Against Acute Alcohol-Induced Gastric Mucosal Injury In Rats
. Elsevier. 1142:6662-6666. Khanza, Abu. 2010. Fit and Fresh through Habbatussauda. Cicero Publishing:
Jakarta. Hal. 45-56. Karmen Gama Baratawidjaja, 2009. Imunologi Dasar Edisi Delapan. Jakarta:
Balai Penerbit FKUI. Kresno, S.B. 1996. Imunologi: Diagnosis dan Prosedur Laboratorium, Ed. III,
Fakultas Kedokteran Universitas Indonesia. Kulisic Z, Tambur Z, Maličević Z, Bakrač NA, Misic Z. 2006. White Blood Cell
Differential Count In Rabbits Artificially Infected With Intestinal Coccidia . J.
Protozool. Res. 16, 42-50. Luo Y, Su Y, Shen Y, Zhao L, Li K. 2004. The Levels of Plasma IL-
1β, IL-6 of C57BL6J Mice Treated with MPTP and Brain Lateralization. Department of
Microbiology and Immunology, Shantou University Medical College, Shantou, China.
Manu KA and Kuttan G. 2008. Immunomodulatory activities of Punarnavine, an alkaloid from Boerhaavia diffusa.
Immunopharmacology and Immunotoxicology: 377–387
Madigan, M.T., J.M. Martinko, dan J. Parker. 2003. Brock Biology of Microorganisms
. Pearson Education, Inc. New Jersey. Halaman 79-80. Mbarek, L. Ait. dkk. 2007. Anti-tumor Properties of Blackseed Nigella sativa l.
Extracts . Cadi-Ayyad University. 40: 839-847
Musa, Daoud. dkk. 2004. Antitumor Activity Of An Ethanol Extract Of Nigella Sativa Seeds
. Harran university. 6: 735—740.
UIN Syarif Hidayatullah Jakarta
Michel CG, dkk. 2010. Phytochemical And Biological Investigation Of The Extracts Of Nigella sativa L. Seed Waste
. Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Egypt
Nakae S, Asano M, Horai R, Iwakura Y. Interleukin- 1β, bt not interleukin-1α is
required for T-Cell- dependent antibody production. Immunology 2001; 104: 402- 409.
Norsharina, ismail. dkk. 2011. Thymoquinone Rich Fraction From Nigella sativa And Thymoquinone Are Cytotoxic Towards Colon And Leukemic Carcinoma Cell
Lines . Academic Journals. pp. 3359-3366.
Omar EM. 2001. Analysis of Single Nucleotide in the Interleukin- 1β Gene Using
5 Nuclease Assays . In: Interleukin Protocols. New Jersey : Humana Press
Paarakh, Padmaa M. 2010. A comprehensive review Nigella sativa Linn. Department of Pharmacognosy, The Oxford College of Pharmacy. pp.409-429.
Padhye, S., Banerjee, S. 2008. Therapeutic Potential Of Black Cumin Seeds And
Beyond . Department of Pathology and Division of Internal Medicine. 6b: 495–
510. Peter, K. V. 2004. Handbook of Herbs and Spices Volume 2. Cambridge England.
Woodhead Publishing Ltd. RAR 2007. Reference values for laboratory animals: normal hematological
values . Research Animal Resources, University of Minnesota.
Salem Mohamed L. 2005. Immunomodulatory and therapeutic properties of the Nigella sativa L. seed
. Int. J. Immunopharmacol: 1749– 1770. Santoso singgih. 2008. Panduan Lengkap Menguasai SPSS 16. PT. Elex Media
Komputindo. Jakarta: 239-247, 315-319 Shoieb AM, Elgayyar M, Dudrick PS, Bell JL, Tithof PK. In Vitro Inhibition Of
Growth And Induction Of Apoptosis In Cancer Cell Lines By Thymoquinone . Int J
Oncol 2003; 22: 107–13.
UIN Syarif Hidayatullah Jakarta
Singh, gurdip. dkk. 2005. Chemical constituents and antimicrobial and antioxidant potentials of essential oil and acetone extract of Nigella sativa
seeds. Chemistry Department, DDU Gorakhpur University. 85:2297–2306.
Sloane E. Anatomi dan Fisiologi. Jakarta: EGC Penerbit Buku Kedokteran. Sutrisno, BB. 1986. Analisis Jamu Edisi I. Fakultas Farmasi Universitas
Pancasila: Jakarta. Hal. 6-9. Suhatri, Aldi Y. 2010. Aktifitas Ekstrak Etanol Biji Jintan Hitam Nigella sativa
Linn. Terhadap Titer Antibodi Dan Jumlah Sel Leukosit Pada Mencit Putih Jantan
. Fakultas Farmasi Universitas Andalas. Sukowati S, 2010. Masalah Vektor Demam Berdarah Dengue DBD dan
Pengedaliannya di Indonesia. Buletin Jendela Epidemiologi. Vol 2 Sumartini S, Zuas O, Julismardiany R, Susilawati E. Aplikasi Elisa Kit Untuk
Mendeteksi Adanya Daging Babi Dalam Makanan . Pusat Penelitian Kimia:
Tangerang Swamy S.M.K. 1998. Cytotoxic and Immunopotentiating Effects of Ethanolic
Extract of Nigella sativa L. Seeds. Department of Pharmacology, Faculty of
Medicine, National University of Singapore. 70 2000 1–7. Tahir K, Bakeet D. 2006. A Plea for Urgent Clinical Evaluation of its Volatile
Oil. Department of Pharmacology, College of Pharmacy, King Saud University Riyadh Saudi Arabia: 2 – 3
Tousson E, El-Moghazy E, El-Atrsh E. 2011.The possible effect of diets containing Nigella sativa and Thymus vulgaris on blood parameters and some
organs structure in rabbit. Toxicology and Industrial Health Underwood, JCE.1996. General and Systematic Pathology Second Edition.
Penerjemah: Sarjadi. Jakarta: Penerbit Buku Kedokteran EGC. 190 Virella G. 1997. Introduction to Medical Immunology Fourth Edition. New York:
Marcel Dekker
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Vieira K. 2011. Improving Abnormal Results. University Of Florida College Of Medicine
Yeheya M. 2009. A Review Therapeutic Role of Phrophetic Medicine Habbat El Baraka Nigella sativa
L.. Departement of Pharmacy. 7 9: 1203-1208.
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Lampiran 1
. Surat Keterangan Hewan Uji
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Lampiran 2 . Alat dan Bahan yang digunakan
Hemasitometer
Tabung Tamo
Mikroskop Setrifus
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Differesial Leukosit Counter
Larutan Turk Larutan Giemsa
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Lampiran 3. Alur Penelitian
Jinten hitam berpotensi sebagai anti inflamasi, anti oksidan, antitumor dan beberapa penelitian menunjukan bahwa jinten hitam berkhasiat sebagai
imunomodulator
Dilakukan peneletian untuk membandingkan dan mengetahui efektifitas ekstrak etanol Jinten hitam terhadap jumlah total leukosit mencit dan jumlah IL-
1β pada mencit yang diberikan LPS
Serbuk Jinten hitam
Ekstrak Etanol
Aklitimasi Hewan Uji
Pemberian ekstrak etanol selama 14 hari berturut -turut
Hari ke – 5 dua jam setelah ekstrak etanol, diberikan LPS 20 μgkgBB
mencit. Enam jam kemudian diambil darah melalui retro-orbital plexus
Pengambilan darah melalui retro-orbital plexus pada hari ke 7, hari ke 14 dan hari
ke 21 Pemberian ekstrak etanol selama 5 hari
berturut -turut
Perhitungan Total Leukosit, Limfosit dan Monosit
Penentuan kadar IL- 1β menggunakan
ELISA
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Lampiran 4. Pembuatan Larutan Uji
A. Pembuatan Larutan Uji Ekstrak Etanol Untuk Uji Total Leukosit
1. Dosis Rendah 2.5 mg20 gr BB atau 125 mgkg BB
VaO Mencit ∶
Dosis �
mg kg
BB �
×
Berat Badan kg Konsentrasi
0.5 ml ∶
125 mg
kg ×0.02kg
Konsentrasi Konsetrasi
: 5 mgml VaO Total
: VaO x Jumlah Mencit x Lama Pemberian : 0.5 ml x 8 x 14
: 56 ml x 2 : 112 ml
Jumlah ekstrak yang ditimbang : VaO Total x Konsentrasi
: 112 ml X 5 mgml : 560 mg
2. Dosis Sedang 5 mg 20 grBB atau 250 mgkgBB
VaO Mencit ∶
Dosis �
mg kg
BB � × Berat Badan kg
Konsentrasi
0.5 ml ∶
250 mg
kg ×0.02kg
Konsentrasi Konsetrasi
: 10 mgml VaO Total
: VaO x Jumlah Mencit x Lama Pemberian : 0.5 ml x 8 x 14
: 56 ml x 2 : 112 ml
Jumlah ekstrak yang ditimbang : VaO Total x Konsentrasi
: 112 ml X 10 mgml : 1120 mg
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3. Dosis Tinggi 10 mg20 grBB atau 500 mgkgBB
VaO Mencit ∶
Dosis �
mg kg
BB � × Berat Badan kg
Konsentrasi
0.5 ml ∶
500 mg
kg ×0.02kg
Konsentrasi Konsetrasi : 20 mgml
VaO Total : VaO x Jumlah Mencit x Lama Pemberian
: 0.5 ml x 8 x 14 : 56 ml x 2
: 112 ml Jumlah ekstrak yang ditimbang
: VaO Total x Konsentrasi : 112 ml X 20 mgml
: 2240 mg
B. Pembuatan Larutan Uji Ekstrak Etanol Untuk Uji Interleukin 1 Beta
1. Dosis Rendah 2.5 mg20 gr BB atau 125 mgkg BB
VaO Mencit ∶
Dosis �
mg kg
BB � × Berat Badan kg
Konsentrasi
0.5 ml ∶
125 mg
kg ×0.02kg
Konsentrasi Konsetrasi
: 5 mgml VaO Total
: VaO x Jumlah Mencit x Lama Pemberian : 0.5 ml x 5 x 14
: 12.5 x 2 : 25 ml
Jumlah ekstrak yang ditimbang : VaO Total x Konsentrasi
:25 ml X 5 mgml :125 mg
2. Dosis Sedang 5 mg 20 grBB atau 250 mgkgBB
VaO Mencit ∶
Dosis �
mg kg
BB � × Berat Badan kg
Konsentrasi
0.5 ml ∶
250 mg
kg ×0.02kg
Konsentrasi
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Konsetrasi : 10 mgml
VaO Total : VaO x Jumlah Mencit x Lama Pemberian
: 0.5 ml x 5 x 14 : 12.5ml x 2
: 25 ml Jumlah ekstrak yang ditimbang
: VaO Total x Konsentrasi : 25 ml X 10 mgml
: 250 mg
3. Dosis Tinggi 10 mg20 grBB atau 500 mgkgBB
VaO Mencit ∶
Dosis �
mg kg
BB � × Berat Badan kg
Konsentrasi
0.5 ml ∶
500 mg
kg ×0.02kg
Konsentrasi Konsetrasi
: 20 mgml VaO Total
: VaO x Jumlah Mencit x Lama Pemberian : 0.5 ml x 5 x 14
: 12.5 :25 ml
Jumlah ekstrak yang ditimbang : VaO Total x Konsentrasi
: 25 ml X 20 mgml : 500 mg
Lampiran 5.
Kegiatan Penelitian
Apusan Darah Fiksasi
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Lampiran 6
. Gambar Pemeriksaan Total Leukosit, Monosit dan Limfosit
Monosit
Limfosit
Total Leukosit
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Lampiran
7. Jumlah Total Leukosit, Presendase Monosit Dan Limfosit Hari 7
1. Jumlah Total Leukosit Pengambilan Hari 7
Kelompok No
Total Lekosit Rata - Rata Total Leukosit mm
3
I II
Kontrol
1 112
91 101.5
5075 2
59 86
72.5 3625
5 64
83 73.5
3675 7
37 110
73.5 3675
8 93
117 105
5250
Rata - Rata 85.2
4260.0
Etanol Dosis Rendah
1 83
85 84
4200 2
167 131
149 7450
3 78
99 88.5
4425 4
185 217
201 10050
7 107
100 103.5
5175
Rata - Rata 125.2
6260.0
Etanol Dosis Sedang
1 162
172 167
8350 2
158 182
170 8500
4 134
140 137
6850 7
144 140
142 7100
8 148
130 139
6950
Rata - Rata 151.0
7550.0
Etanol Dosis Tinggi
1 211
178 194.5
9725 2
285 278
281.5 14075
3 192
150 171
8550 4
241 203
222 11100
6 250
234 242
12100
Rata - Rata 222.2
11110.0
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2. Persentase Monosit dan Limfosit Hari 7
Kelompok No
Differensial Lekosit Jumlah Differensial Leukosit
Monosit Limfosit Eusonofil Basofil Monosit Limfosit Eusonofil Basofil
Kontrol 1
8 86
1 5
406 4364.5
50.75 253.75 2
9 85
1 6
326.25 3081.25 36.25
217.5 5
8 89
2 2
294 3270.75 73.5
73.5 7
4 89
2 5
147 3270.75 73.5 183.75
8 1
95 2
2 52.5
4987.5 105
105
Rata - Rata 6
89 2
4 245.2
3795.0 67.8
166.7
Etanol Dosis Rendah
1 1
99 42
4158 2
5 94
1 372.5
7003 74.5
3 6
94 265.5
4159.5 4
11 88
1 1105.5
8844 100.5
7 12
88 621
4554
Rata - Rata 7
93 481.3
5743.7 20.1
14.9
Etanol Dosis Sedang
1 4
96 334
8016 2
8 90
2 680
7650 170
4 4
95 1
1 274
6507.5 68.5
68.5 7
3 96
1 213
6816 71
8 5
95 347.5
6602.5
Rata – Rata 5
94 1
369.7 7118.4
13.7 61.9