PENGARUH KEFIR KOMBINASI SUSU KAMBING DAN SUSU KEDELAI TERHADAP KADAR TUMOR NECROSIS FACTOR ALPHA (TNF-α) PLASMA DAN EKSPRESI GEN INDUCIBLE NITRIC OXIDE SYNTHASE (iNOS) HEPAR PADA TIKUS YANG DIINDUKSI STREPTOZOTOCIN DAN NICOTINAMIDE

PRANITA ARITRINA, Dr. Sunarti, M.Kes.

2015 | Tesis | S2 Ilmu Kedokteran Dasar dan Biomedis

Latar Belakang. Hiperglikemia kronik pada Diabetes Melitus (DM) dapat meningkatkan produksi sitokin proinflamasi. Hal ini menyebabkan destruksi sel beta pankreas dan disfungsi endokrin pankreas. Kefir merupakan salah satu jenis susu fermentasi yang dilaporkan memiliki efek antidiabetes, antioksidan, dan antiinflamasi karena biji kefir mengandung bakteri asam laktat dan yeast. Kefir dapat dibuat dari inokulasi berbagai jenis susu seperti susu sapi, susu kambing, maupun susu kedelai dengan biji kefir. Tujuan. Penelitian ini bertujuan untuk mengetahui pengaruh kefir kombinasi susu kambing dan susu kedelai terhadap kadar TNF-α plasma dan ekspresi gen iNOS hepar pada tikus yang diinduksi streptozotocin (STZ) – nicotinamde (NA). Metode. Jenis penelitian ini adalah ekperimental kuasi dengan rancangan post-test only control group design. Subjek penelitian adalah tikus Wistar jantan, diinduksi STZ-NA, diberi perlakuan kefir susu kambing, kefir susu kedelai, dan kefir kombinasi susu kambing dan susu kedelai. Kadar Glukosa Darah Puasa (GDP) plasma diperiksa dengan metode enzymatic photometric, kadar TNF- α plasma diperiksa dengan metode ELISA, dan ekspresi gen iNOS diperiksa dengan metode Real Time-PCR. Hasil. Kadar TNF- α plasma tertinggi ditemukan pada kelompok tikus kontrol DM dan terendah pada kelompok tikus DM yang diberi perlakuan kefir kombinasi susu kambing dan susu kedelai. Kadar TNF-α plasma pada kelompok tikus DM yang diberi kefir susu kambing, kefir susu kedelai, dan kefir kombinasi susu kambing dan susu kedelai lebih rendah dibandingkan kelompok tikus kontrol DM dan kontrol sehat. Hasil uji Kruskal-Wallis menunjukkan bahwa terdapat perbedaan bermakna kadar TNF-α plasma antar kelompok (p ≤ 0,05). Hasil uji Mann-Whitney menunjukkan bahwa terdapat perbedaan bermakna (p ≤ 0,05) kadar TNF-α plasma kelompok tikus kontrol DM dengan kelompok DM yang diberi kefir, sedangkan kadar TNF-α antar 3 kelompok DM yang diberi kefir tidak menunjukkan perbedaan bermakna (p ≥ 0,05). Ekspresi gen iNOS relatif terhadap kontrol sehat paling tinggi ditemukan pada kelompok tikus DM yang diberi kefir susu kedelai dan paling rendah ditemukan pada kelompok tikus DM yang diberi kefir kombinasi susu kambing dan susu kedelai. Ekspresi gen iNOS relatif pada kelompok tikus kontrol DM lebih tinggi dibandingkan dengan kelompok tikus DM yang diberi kefir susu kambing maupun kefir kombinasi susu kambing dan susu kedelai. Terdapat perbedaan bermakna ekspresi relatif pada kelompok kontrol sehat, kontrol DM maupun 3 kelompok DM yang diberi perlakuan berupa kefir susu kambing, kefir kombinasi susu kambing dan kedelai, serta kefir susu kedelai berdasarkan uji one way ANOVA (p ≤ 0,05). Kesimpulan. Pemberian kefir kombinasi susu kambing dan susu kedelai dapat menurunkan kadar TNF- α plasma dan menekan ekspresi gen iNOS hepar pada tikus yang diinduksi STZ-NA. Kata Kunci: Diabetes Melitus, Inflamasi, TNF- α, Ekspresi gen iNOS, Kefir Kombinasi Susu Kambing dan Susu Kedelai

Background. Chronic hyperglicemia on Diabetes Mellitus (DM) can increase the production of proinflammatory cytokine. This situation can cause destruction of beta pancreas cell and dysfunction of pancreas endocrine. Kefir is one of fermentation milk which known has an effect for antidiabetic, antioksidan, and antiinflammatory, because kefir seed contents of lactic acid bacteria and yeast. Kefir can be made by inoculation of various kind of milk, such as from cow, goat, or soybean with kefir seed. Purpose. This research aims to know about the impact of combination of goat-milk and soybean-milk kefir on TNF-α plasma level and iNOS gene expression on streptozotocin– nicotinamde induced rats. Methodology. This is a quasi-experimental research with post-test only control group design. Subject in this research is Wistar rats, which induced by STZ-NA, and given treatment by goat-milk kefir, soybean-milk kefir, and combination between goat-milk and soybean-milk kefir. The fasting glucose plasma level was measured by glucose oxidase-peroxidase (GOD-PAP) method, TNF- α plasma level was measured by Enzym Linked Immunosorbent Assay (ELISA) method, and iNOS gene expression was measured by Real Time-PCR method. Results. The Highest level of TNF- α plasma was found in diabetic control rats group, and the lowest was found in diabetic rats with combination of goat-milk and soybean-milk kefir group. Diabetic rats with treated by goat milk kefir, soybean-milk kefir, and combination of goat-milk and soybean-milk kefir has lower level than diabetic control rats and health rats group. The result of Kruskal-Wallis test showed the significant difference of TNF- α plasma level between the groups (p ≤ 0,05). The Mann-Whitney test showed the significant difference (p ≤ 0,05) of TNF-α plasma level between diabetic control rats group and diabetic rats group which treated by kefir, meanwhile TNF-α level between three diabetic rats groups which treated by kefir do not show the significant difference (p ≥ 0,05). The highest level of iNOS gene expression relatively to health-controlled was found in diabetic rats group with soybean-milk kefir and the lowest level was found in diabetic rats group with combination of goat-milk and soybean-milk kefir. iNOS gene expression on diabetic rats control group has higher level than diabetic rats group which treated by goat-milk kefir and combination of goat-milk and soybean-milk kefir. There is significant difference in relative expression on health-group, diabetic control rats group, and 3 diabetic rats groups which had treatment with goat-milk kefir, combination of goat-milk and soybean-milk kefir, and soybean-milk kefir based on one-way ANOVA test (p ≤ 0,05). Conclusion. Combination of goat-milk and soybean-milk kefir can decrease TNF- α plasma level and keep iNOS hepar gene expression on STZ-NA induced rats. Keywords: Diabetes Mellitus, Inflamation, TNF- α, iNOS gene expression, Kefir Combination Goat-Milk and Soybean-Milk

Kata Kunci : Diabetes Melitus, Inflamasi, TNF- α, Ekspresi gen iNOS, Kefir Kombinasi Susu Kambing dan Susu Kedelai; Diabetes Mellitus, Inflamation, TNF- α, iNOS gene expression, Kefir Combination Goat-Milk and Soybean-Milk