Cory’ah, Fitra Arsy Nur (2017) Pengaruh Ekstrak Etanol Pegagan (Centella Asiatica) Terhadap Panjang Badan, Ekspresi Insulin Like Growth Factor-1 (Igf-1) Dan Insulin Reseptor Substrat (Irs) Pada Larva Zebrafish (Danio Rerio) Model Stunting Dengan Induksi Rotenon. Magister thesis, Universitas Brawijaya.
Abstract
Stunting merupakan retardasi pertumbuhan linier (RPL) pada anak balita, karena malnutrisi jangka panjang. Stunting menurut standar baku dari WHO Child Growth Standart tahun 2010, didasarkan pada indikator pengukuran panjang badan dibanding umur atau tinggi badan dibanding umur dengan nilai (z-score) < -2 SD (Standar Deviasi). Riset Kesehatan Dasar 2013 mencatat prevalensi stunting nasional mencapai 37,2 %. Prevalensi stunting di Indonesia lebih tinggi daripada negara-negara lain di Asia Tenggara. Kriteria stunting adalah lahir dalam kondisi normal dan nampak pada usia 2 tahun. Faktor yang berpengaruh terhadap pertumbuhan dan perkembangan adalah faktor genetik dan faktor lingkungan. Faktor lingkungan yang berpengaruh adalah bahan kimia toksik, seperti pestisida atau bahan xenobiotik lainnya. menyebabkan gangguan sintesis, transport, metabolism dan kerja hormon-hormon dalam tubuh. Sehingga meningkatkan resiko abnormal pada pertumbuhan dan perkembangan. Rotenone merupakan pestisida alami memiliki mekanisme kerja yaitu menghambat komplek I mitokondria yang merupakan sumber ROS yang berakibat peningkatan tekanan oksidasi, ketersediaan ATP dan Kematian sel. Mekanisme stunting dapat dikaitkan dengan keadaan malnutrisi, gangguan pada hormon pertumbuhan dan inflamasi. Hormon pertumbuhan berperan menstimulasi pembentukan IGF-1 (Insulin Like Growth Factor-1) sebagai mediator pertumbuhan prenatal dan postnatal, pada otot, tulang rawan, dan diferensiasi tulang lainnya. Sedangkan IRS (insulin reseptor substrat) sebagai efektor, melaksanakan fungsi utama pada reseptor untuk transduksi sinyal, menghasilakn diferensiasi sel, proliferasi dan anti apoptosis, pengaturan metabolisme, umur dan ukuran organisme. Dua periode penting dalam menilai rendahnya linear growth, yaitu gangguan pertumbuhan intrauterine, serta rendahnya kadar IGF-1 dan inflamasi. Centella asiatica salah satu tanaman obat yang memiliki toksisitas rendah dan memiliki kandungan nutrisi lengkap yaitu makronutrien (karbohidrat, dan protein), Mikronutrien (vitamin dan mineral), serta fitonutrien (flavonoid, triterpen, karotenoid, alkaloid dan glikosida). Centella asiatica memiliki manfaat salah satunya mengandung senyawa bioaktif triterpenoid (asiaticoside,medecasossida) yang tinggi, sebagai antioksidan dan antiinflamasi mampu menghambat terjadinya reaksi oksidasi akibat paparan rotenon. Penelitian ini bertujuan untuk membuktikan model stunting larva zebrafish melalui induksi rotenon, dan mengetahui efek pemberian ekstrak Centella asiatica pada model stunting larva zebrafish yang diinduksi rotenone melalui ekspresi IGF-1 dan IRS, serta mengetahui hubungan ekspresi IGF-1 dengan IRS. Metode penelitian ini adalah true eksperimental laboratori in vivo. Desain penelitian randomize posttest only control group design dengan random alokasi. Hewan coba yang digunakan adalah embrio zebrafish (Danio rerio) usia 0-9 dpf (day post fertilization), yang dibagi menjadi lima kelompok yaitu kelompok kontrol negatif (embrio normal); kelompok kontrol positif (embrio yang diinduksi rotenone 12,5 ppb); kelompok perlakuan1,2 dan 3 secara berurutan( rotenon 12,5 ppb ditambah pegagan konsentrasi 1,25 μg/mL, 2,5 μg/mL dan 5 μg/mL). Pemberian rotenone dan pegagan dilakukan pada usia 2 hingga 72 hpf (hour post fertilization). Pengamatan panjang badan dan rasio panjang kepala dilakukan pada 3,6 dan 9 dpf dg software image ruster versi.3 dari optilab versi 2.2.1. Pada usia 9 dpf larva zebrafish diterminasi, selanjutnya dilakukan pengukuran ekspresi IGF-1 dan IRS dengan metode IHC (Immunohistochemistry) wholemount dengan pewarnaan DAB (Diamino benzidine). Selanjutnya dilakukan pengamatan dan kuantifikasi ekspresi melalui integrated density dengan software image ix J versi 1.50. Analisis data menggunakan software SPSS versi 18 dengan uji T-Test dan One Way Anova. Hasil penelitian menunjukan bahwa, pada usia 3 dpf (analog bayi baru lahir) tidak ada perbedaan signifikan panjang badan antara semua kelompok (nilai p Value = 0,385). Sedangkan pada usia 6 dpf (analog usia 2 tahun) dan 9 dpf (analog usia 8 tahun) ada perbedaan signifikan panjang badan yaitu kelompok rotenon lebih pendek dibandingkan dengan kelompok kontrol dan kelompok pegagan (p = 0,000). Ekspresi IGF-1 mengalami penurunan signifikn pada kelompok rotenon (p=0,000), penambahan pegagan mampu meningkatkan ekspresi IGF-1 yang tergantung dosis. Peningkatan signifikan terjadi pada RP5(p=0,000). Adanya kecendrungan peningkatan ekspresi IRS yang diberikan ekstrak pegagan, walaupun tidak berbeda signifikan dengan kelompok rotenon. Korelasi pemberian pegagan dengan IGF-1 didapatkan hubungan yang sangat kuat (p=0,728). Korelasi pemberian pegagan dengan IRS didapatkan hubungan yang lemah (p=0,397). Dapat disimpulkan bahwa rotenon 12,5 ppb mampu menginduksi stunting pada larva zebrafish. Derajat kepercayaan 98%. Pemberian Centella asiatica mampu meningkatkan pertumbuhan panjang badan larva zebrafish melalui peingkatan ekspresi IGF-1 serta IRS pada konsentrasi 5 μg/mL.
English Abstract
Stunting is linear growth retardation (RPL) in children under five, due to long-term malnutrition. Stunting according to WHO Child Growth Standard 2010, is based on an indicator of body length over age or height over age with a z-score<-2 SD (Standard Deviation). Primary Health Research 2013 records the national stunting prevalence is 37.2%. The Stunting criterion is born normally and it detected at 2 years old. The prevalence of stunting in Indonesia is higher than other countries in Southeast Asia. Factors that affect growth and development are genetic and environmental factors. An influential environmental factor is toxic chemicals, such as pesticides or other xenobiotic substances.Those cause disruption of synthesis, transport, metabolism and the work of hormones in the body. Therefore, it increases the risk of abnormal growth and development. Rotenone is a natural pesticide which mechanism inhibits complex I mitochondria,increasing of ROS, oxidative stress occurs, failure of production and cell death. Stunting mechanisms may be associated with malnutrition insuficiency of growth hormone and inflammatory disorders. Growth hormone stimulates the formation of IGF-1 (Insulin like Growth Factor-1) as a mediator of prenatal and postnatal growth, in muscle, cartilage, and other bone differentiation. While the IRS (insulin receptor substrate) as an effector performs major functions on receptors for signal transduction, produces cell differentiation, proliferation and anti-apoptosis, metabolic regulation, age and size of organism. The important periods in assessing low linear growth, i.e. intrauterine growth disorder, and low levels of IGF-1 and inflammation as well. Centella asiatica one of the medicinal plants with low toxicity and have a complete nutrient content including macronutrient (carbohydrate, and protein), micronutrients (vitamins and minerals) and phytonutrients (flavonoids, triterpenes, carotenoids, alkaloids and glycosides). Centella asiatica has other benefits of due to its bioactive compounds such as triterpenoids (asiaticoside, medecasossida) which has powerful anti-inflammatory and antioxidant that able to inhibit oxidative stress by otenone exposure.This study aims to proved rotenone-induced zebrafish larvae stunting model through rotenone induction, observed the effects of Centella asiatica extract on rotenone-induced zebrafish larvae stunting model through the expression of IGF-1 and IRS, as well as determine the relationship of the expression of IGF-1 with the IRS. This research method was true experimental laboratory in vivo. Research design randomized posttest only control group design with random allocation. The experimental animals used zebrafish (Danio rerio) ages 0-9 dpf, divided into five groups: the negative control group (normal embryo); Positive control group (rotenone induced embryo 12.5 ppb); Group of treatment were rotenone plus pegagan ,25 μg / mL, 2.5 μg / ml and 5 μg / L respectively. Administration of rotenone and pegagan was done at the age of 2 to 72 hpf. Body length and head length ratio were performed at 3.6 and 9 dpf with image ruster version.3 software from optilab version 2.2.1. At 9 dpf zebrafish larvae were terminated, subsequent measurement of the expression of IGF-1 and IRS by methods IHC (Immunohistochemistry) wholemount and stained with DAB (diaminobenzidine). Furthermore, the observation and quantification of expression through integrated density with image software J version 1.50. The analysis of data used SPSS software version 18 by through T-Test and One Way Anova analysis. The results showed that, at 3 dpf (new born baby) there was no significant difference in body length among the groups (p=0.113). While at 6 dpf (analog to 2 years old) and 9 dpf (analog to 8 years old) there was a significant difference in length of the shorter rotenone group compared to the control group and pegagan group (p = 0.000). xi Expression of IGF-1 significantly decreased on rotenone group (p = 0.000), the administration of pegagan could was the shortest length, increase the expression of IGF-1 depend on the dose, RP5 was measured significantly (p = 0.000). There was IRS expression was increased by administration of pegagan, but not significantly measured. The correlation of pegagan administration with IGF-1 was found to be very strong (p = 0.728). The correlation of pegagan administration with IRS was found to be weak relationship (p = 0,397). It concluded that 12.5 ppb rotenone is could of induce stunting in zebrafish larvae. The degree of confidence is 98%. The administration of Centella asiatica was able to increase the growth of zebrafish larvae body length by increasing the expression of IGF-1 and IRS at 5 μg / mL concentration.
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/615.323 849/COR/p/2017/041708996 |
| Uncontrolled Keywords: | UMBELLIFERNE, ZEBRA DANIO, INSULIN - LIKE GROWTH FACTOR - BINDING PROTEINS, ROTENONE |
| Subjects: | 600 Technology (Applied sciences) > 615 Pharmacology and therapeutics > 615.3 Organics drugs > 615.32 Drugs derived from plants and mikroorganisms > 615.323 87 Drugs derived from olive oil |
| Divisions: | S2/S3 > Magister Kebidanan, Fakultas Kedokteran |
| Depositing User: | Nur Cholis |
| Date Deposited: | 03 Oct 2017 06:09 |
| Last Modified: | 03 Jun 2024 07:11 |
| URI: | http://repository.ub.ac.id/id/eprint/3181 |
Preview |
Text
BAGIAN DEPAN.pdf Download (979kB) | Preview |
Preview |
Text
BAB I .pdf Download (180kB) | Preview |
Preview |
Text
BAB II .pdf Download (1MB) | Preview |
Preview |
Text
BAB III .pdf Download (184kB) | Preview |
Preview |
Text
BAB IV .pdf Download (416kB) | Preview |
Preview |
Text
BAB V .pdf Download (416kB) | Preview |
Preview |
Text
BAB VI .pdf Download (223kB) | Preview |
Preview |
Text
BAB VII .pdf Download (146kB) | Preview |
Preview |
Text
DAFTAR PUSTAKA.pdf Download (288kB) | Preview |
Actions (login required)
 |
View Item |