Pengaruh Pemberian Nanoemulsi Pentagamavunon-0 terhadap Ekspresi Gen Histone Deacetylase 2, Gen Bdnf, Gen Neurogenesis dan Gliogenesis pada Mencit Model Brain Disorder serta Karakterisasinya

VANIA AMANDA SAMOR, Dr. apt. Nunung Yuniarti, M.Si. ; Dr. apt. Muthi Ikawati, M.Sc.

2022 | Tesis | MAGISTER ILMU FARMASI

Neural Stem Cells (NSC) merupakan cikal bakal sel neuron dan sel glia yang proses diferensiasinya, yaitu neurogenesis dan gliogenesis, dilaporkan dipengaruhi oleh keseimbangan aktivitas enzim histone acetyltransferase (HAT) dan histone deacetylase (HDA). Gangguan keseimbangan ekspresi kedua gen tersebut berperan dalam penyakit neurodegeneratif seperti penyakit Alzheimer. Studi in silico mengungkap bahwa kurkumin dan turunannya, Pentagamavunon-0 (PGV-0) merupakan ligan HDA2 dan HDA8 yang poten. Studi in vitro dan in vivo melaporkan bahwa PGV-0 menghambat HDA2 dan meningkatkan learning and memory, interaksi sosial, serta menurunkan kecemasan pada hewan uji. Penelitian ini bertujuan untuk menelusuri potensi PGV-0 dalam formulasi self-nanoemulsifying drug delivery system (SNEDDS) sebagai solusi permasalahan bioavailibilitas PGV-0, melalui kaitannya pada ekspresi Hda2, Bdnf sebagai marker plastisitas NSC, serta gen-gen yang terkait neurogenesis (Neun, Map2, dan Tuj1) dan gliogenesis (Gfap, S100beta, Olig2). Karakterisasi SNEDDS PGV-0 dilakukan untuk melihat distribusi ukuran partikel dan muatannya menggunakan particle size analyzer (PSA). Induksi brain disorder, dilakukan pada mencit jantan galur Balb/c dengan etanol 10% v/v dalam CMC-Na 0,5% dilanjutkan dengan pemberian suspensi/SNEDDS PGV-0 dosis 20 atau 40mg/kgBB selama 21 hari secara per oral. mRNA diisolasi dari sampel otak dan ekspresi gen diamati menggunakan reverse transcriptase-PCR dan qPCR untuk Bdnf, kemudian divisualisasi dengan gel elektroforesis. Hasil pita dianalisis secara densitometri dengan ImageJ dan dinormalisasi dengan Gapdh. SNEDDS (20 dan 40) mg/kgBB memiliki indeks polidispersitas berturut-turut (0.279 +/- 0.00 nm dan 0.104 +/- 0.01), ukuran partikel (11.75 +/- 0.03 nm dan 12.05 +/- 0.03) nm dan potensial zeta (6.98 +/- 0.36 dan 6.08 +/- 0.10) mV. Pemberian SNEDDS PGV-0 20 dan 40 mg/kgBB tidak menghambat ekspresi Hda2 namun mengembalikan level Hda2 pada level normal, meningkatkan neurogenesis melalui Bdnf, NeuN, Map2 dan gliogenesis melalui S100beta, dan Olig2, tetapi tidak mempengaruhi Tuj1 dan Gfap. SNEDDS PGV-0 20 dan 40 mg/kgBB berpotensi sebagai agen brain-disorder berdasarkan ekspresi gen tersebut.

Neural Stem Cells (NSC) are the precursors of neurons and glial cells, which then known as neurogenesis and gliogenesis, respectively. Neurogenesis and gliogenesis are said to be orchestrated by the balance of histone acetyltransferase (HAT) and histone deacetylase (HDA) enzyme activities. Disruption of these enzymes reportedly play a role in neurodegenerative diseases such as Alzheimer disease. An in silico study revealed that curcumin and its derivative, Pentagamavunon-0 (PGV-0), are effective as HDA2 and HDA8 ligands. PGV-0 inhibits HDA2 and improves learning and memory, social interaction, and anxiety in animals, according to previous in vitro and in vivo studies. This study aims to explore the potential of PGV-0 in the formulation of a self-nanoemulsifying drug delivery system (SNEDDS) as a solution to the PGV-0 bioavailability problem, by examining its relationship to the expression of Hda2, Bdnf as a marker of NSC plasticity, and genes related to neurogenesis (Neun, Map2, and Tuj1) and gliogenesis (Gfap, S100beta, and Olig2). The characterization of SNEDDS PGV-0 was carried out to see the distribution of particle size and charge using particle size analyzer (PSA). Brain disorder induction was carried out in Balb/c strain male mice using 10% v/v ethanol in 0.5% CMC-Na, followed by oral gavage administration of suspension/SNEDDS PGV-0 at doses of 20 or 40 mg/kgBB for 21 days. mRNAs were isolated from brain samples, and gene expression was analyzed with reverse transcriptase-PCR and qPCR for Bdnf, which then visualized using gel electrophoresis. The band results were analyzed by densitometry using ImageJ and normalized with Gapdh. SNEDDS (20 and 40) mg/kgBW polydispersity index were (0.279 +/- 0.00 nm and 0.104 +/- 0.01), particle size were (11.75 +/- 0.03 nm and 12.05 +/- 0.03) nm and zeta potential were (6.98 +/- 0.36 and 6.08 +/- 0.10) mV respectively. Administration of SNEDDS PGV-0 20 and 40 mg/kgBW did not restrict Hda2 expression but at the same time restored Hda2 levels compared to normal group, increased neurogenesis via Bdnf, NeuN, Map2 and also increased gliogenesis via S100beta, and Olig2, but did not increase Tuj1 and Gfap gene expressions. SNEDDS PGV-0 20 and 40 mg/kgBW are potential as agents of brain disorders based on the expression of these genes.

Kata Kunci : Pentagamavunon-0 (PGV-0), self-nanoemulsifying drug delivery system (SNEDDS), histone deacetylase 2, brain disorder, neurogenesis