Norisa, Nanda (2019) Pengaruh Ekstrak Etanolik Pegagan (Centella asiatica) terhadap Ekspresi Dopamin Reseptor D1, Ekspresi Dopamin Reseptor D2, dan Aktivitas Lokomotor pada Embrio Zebrafish yang Dipapar AlCl3. Magister thesis, Universitas Brawijaya.
Abstract
Aluminium merupakan salah satu jenis logam berat yang paparannya dapat berefek pada gangguan fungsi otak, paru-paru, ginjal, dan organ penting lainnya. Paparan aluminium secara terus-menerus selama masa kehamilan dan menyusui mengakibatkan gangguan perkembangan persisten seperti cacat pada refleks motorik-sensorik, aktivitas lokomotor, perilaku, dan kognitif. Aluminium sangat neurotoksik dan dapat menghambat perkembangan otak prenatal dan postnatal. Paparan aluminium di otak dapat meningkatkan reactive oxygen species (ROS) sehingga terjadi stres oksidatif. Peningkatan ROS tanpa diimbangi dengan peningkatan antioksidan akan menurunkan sintesis dopamin dengan mengganggu aktivitas tirosin hidroksilase. Penurunan dopamin mempengaruhi ekspresi dopamin reseptor D1 (DRD1) dan dopamin reseptor D2 (DRD2) yang menyebabkan gangguan aktivitas lokomotor. Pegagan (Centella asiatica) merupakan salah satu tumbuhan yang memiliki efek antioksidan yang mampu mencegah stres oksidatif serta membersihkan radikal bebas. Tujuan dari penelitian ini adalah untuk membuktikan pengaruh ekstrak etanolik pegagan (Centella asiatica) terhadap ekspresi DRD1 dan DRD2, dan aktivitas lokomotor pada embrio zebrafish yang dipapar AlCl3. Desain penelitian adalah true experimental-post test only control group, 90 embrio zebrafish dibagi dalam 5 kelompok yaitu kelompok kontrol negatif, kelompok AlCl3 50 μM, kelompok AlCl3 ditambah pegagan 1,25 μg/mL, 2,5 μg/mL, dan 5 μg/mL, perlakuan diberikan mulai 2-72 hpf (hour post fertilization), kemudian diukur aktivitas lokomotornya pada usia 4-6 dpf (day post fertilization) dengan cara menghitung pergerakan larva melewati garis kemudian membuat pola pergerakannya dengan menggunakan ImageJ. Pada usia 6 dpf diperiksa ekspresi DRD1 dan DRD2 dengan metode Reverse Transcription Polymerase Chain Reaction (RT-PCR) kemudian di kuantifikasi dengan Gel Doc. Penelitian menghasilkan bahwa paparan AlCl3 menyebabkan terjadinya penurunan ekspresi DRD1 yang tidak berbeda signifikan (p > 0.05) antar kelompok, namun terdapat kecenderungan kelompok yang dipapar AlCl3 memiliki rerata paling rendah bila dibandingkan kelompok kontrol negatif dan kelompok perlakuan. Pemberian ekstrak etanolik pegagan dengan konsentrasi berbeda membuktikan bahwa semakin tinggi konsentrasi pegagan, semakin meningkat ekspresi DRD1. Sebaliknya akibat paparan AlCl3 menyebabkan peningkatan ekspresi DRD2 yang tidak berbeda signifikan (p > 0.05) antar kelompok, namun terdapat kecenderungan kelompok yang dipapar AlCl3 memiliki rerata paling tinggi bila dibandingkan kelompok lain. Pemberian ekstrak etanolik pegagan dengan konsentrasi berbeda membuktikan bahwa semakin tinggi konsentrasi pegagan semakin menurunkan ekspresi DRD2. Aktivitas lokomotor menunjukkan adanya perbedaan signifikan (p < 0.05), pemberian ekstrak etanolik pegagan meningkatkan aktivitas lokomotor seiring peningkatan konsentrasi. Korelasi Spearman antara ekspresi DRD1 dengan ekspresi DRD2 menunjukkan hubungan berlawanan (negatif) sangat kuat (1.000), artinya semakin tinggi ekspresi DRD1 maka semakin rendah ekspresi DRD2. Ekspresi DRD1 dengan aktivitas lokomotor menunjukkan hubungan searah (positif) sangat kuat (1.000), artinya semakin tinggi ekspresi DRD1 maka aktivitas lokomotor semakin meningkat. Ekspresi DRD2 dengan aktivitas lokomotor menunjukkan hubungan berlawanan (negatif) sangat kuat, artinya semakin tinggi ekspresi DRD2 maka aktivitas lokomotor semakin menurun. Kesimpulan pada penelitian ini adalah ekstrak etanolik pegagan (Centella asiatica) dapat meningkatkan ekspresi DRD1, menurunkan ekspresi DRD2, dan meningkatkan aktivitas lokomotor pada embrio zebrafish yang dipapar AlCl3.
English Abstract
Aluminum is a type of heavy metal that can be accessed with an effect on brain, lung, kidney and other important organs. Intensive exposure to aluminum during pregnancy and lactation might lead to persistent developmental disorders such as defects in sensory-motor reflexes, locomotor activity, behavior, and cognitive. Aluminum is highly neurotoxic and it can inhibit prenatal and postnatal brain development. Aluminum exposure in the brain increases the reactive oxygen species (ROS) which leads to oxidative stress. Increases in ROS without increases in antioxidants will reduce dopamine synthesis by disrupting the tyrosine hydroxylase activity. Low level of dopamine will also affect dopamine receptor D1 (DRD1) and dopamine receptor D2 (DRD2) expression, leading to disruptions in locomotor activity. Centella asiatica contains antioxidant agents that can prevent oxidative stress from occurring and help cleanse the free radicals. This research aimed to examine the effect of ethanolic extract from Centella asiatica on DRD1 and DRD2 expression, and locomotor activity in zebrafish embrio exposed to AlCl3. This research designed true experimental of post-test only control group design, 90 zebrafish embryos were divided into 5 groups, negative control group, AlCl3 group 50 μM, AlCl3 group treated with Centella asiatica extract 1.25 μg/mL, 2.5 μg/mL, and 5 μg/mL concentrations, the treatments were started at 2-72 hpf (hour post fertilization). The locomotor activity was measured at 4-6 dpf (day post fertilization) by calculating the movement of larvae over the line and creating movement pattern using ImageJ. At 6 dpf, the DRD1 and DRD2 expression was examined using the Reverse Transcription Polymerase Chain Reaction (RT-PCR) method and quantified using Gel Doc. The study resulted that exposure to AlCl3 caused decreased in DRD1 expression at insignificant level (p> 0.05) between group. However, there was a tendency for groups exposed to AlCl3 to have lower average value than to the negative control group and the treatment group. Different concentrations of Centella asiatica ethanolic extract in this research showed that higher concentration led to higher DRD1. Conversely, exposure to AlCl3 increased the DRD2 expression at insignificant rate (p> 0.05) between groups. However, groups exposed to AlCl3 showed a tendency to have higher average value than other groups. The higher the concentration of Centella asiatica, the lower the expression of DRD2. Locomotor activity showed a significant difference (p <0.05), in which the ethanolic extract from Centella asiatica increased the locomotor activity as concentration increased. A highly strong negative correlation (1.000) was found between DRD1 expression and the DRD2 expression based on the Spearman Correlation Test. It means that the higher the DRD1 expression, the lower the DRD2 expression. The correlation between DRD1 expression and locomotor activity was found highly strong and positive (1,000), meaning that the higher the DRD1 expression, the greater the locomotor activity. Furthermore, the correlation between DRD2 expression and locomotor activity was a strong negative correlation, showing that higher DRD2 expression would lead to lower locomotor activity. This research confirmed that ethanolic extract Centella asiatica could increase expression of DRD1, decrease expression of DRD2, and increase the locomotor activity on AlCl3-induced zebrafish embryo.
Other obstract
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| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/615.107 24/NOR/p/2019/041906244 |
| Uncontrolled Keywords: | MEDICINE, EXPERIMENTAL, MEDICINAL PLANTS, HERBS--THERAPEUTIC USE, DOPAMINE—RECEPTORS, ALUMINUM CHLORIDE |
| Subjects: | 600 Technology (Applied sciences) > 615 Pharmacology and therapeutics > 615.1 Drugs (materia medica) > 615.107 24 Experimental research |
| Divisions: | S2/S3 > Magister Kebidanan, Fakultas Kedokteran |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 18 Dec 2019 07:49 |
| Last Modified: | 21 Oct 2021 06:22 |
| URI: | http://repository.ub.ac.id/id/eprint/177103 |
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