Analysis of the Histopathology, TNF-α of Microglia Cells Expression, NRG-1/erbB Oligodendrocyte, and Ki67/Apoptosis of Dentate Gyrus Rattus novergicus Brain After Acute Traumatic Brain Injury

Head trauma or traumatic brain injury (TBI) gives most serious impact on the central nervous system. Several experimental models have been established to mimic different pathogenesis characteristics of TBI. The purpose of this study was to determine whether there is evidence of hystopathological lesions in the brain tissue after Marmorou TBI models. This study uses Rattus norvegicus Sprague Dawley strain. Macroscopic and microscopic observations on the brain tissue were done. Macroscopic lesions were observed in the brain. Microscopic observation was performed with Haematoxylin-Eosin (HE) staining and immunohistochemistry on the distribution of microglia cells and pyramidal cells in the cortex. Meanwhile, the distribution of NRG-1/ErbB, proliferation, and apoptosis were observed in the hippocampus. The results of macroscopic observation showed that there were wounds caused by falling loads and vasodilatation. On microscopic observation, the TBI group showed an increase in neutrophils distribution and distribution of activated microglia to produce TNF-α, and decrease in the number of cortical pyramidal cells significantly. The distribution of NRG-1 tended to decrease after exposure of TBI and had no effect on its receptor, erbB. Exposure of TBI appears to lower the activity of neuronal cells proliferation in dentate gyrus (DG) area and significantly increase the number of apoptotic cells. Marmarou model is a physiological model of TBI that spontaneously occurs following a trauma to the head, for example trauma due to an accident. This data can be used as a preliminary data of inflammation and tissue regeneration of disrupted adult brain. Therefore, this research could be used as the basis in the studies of therapeutic agents in the process of neurogenesis of brain cells.Keywords: traumatic brain injury, ERG-1/ErbB, dentate gyrus, Ki67, TNF-a, microgli

Spade Leaf Extract Phytosome Modulates Krox-20, Neuregulinـ1, Phospholipids, and Cognitive Function of Traumatic Brain Injury Model in Rats

Traumatic brain injury (TBI) is a disorder of the central nervous system due to head trauma. TBI can damage nerve membrane phospholipids and decrease protein synthesis of neuregulin-1 (NRG-1) because of the transcription factor Krox-20. These conditions cause the lowering in nerve re-myelination which contribute to the decline of cognitive function. In Indonesia, citicoline is a neuroprotective drug that widely used to repair and prevent further damage of the nerve cells membrane caused by trauma. Spade leaf (Centella asiatica) extract phytosome (SEP) is a model of drug delivery system which expected to enhance the therapeutic effects as neuroprotective drug. This study aims to demonstrate and compare the effectiveness from SEP and citicoline as a neuroprotective characterized by increasing the activation of Krox-20, the expression of NRG-1, the distribution of phospholipids, and the improvement of cognitive levels on TBI-induced rats. Rats were divided into 5 groups namely: control (-); control (+); treatment with SEP 90mg/kgBW; citicoline 250mg/kgBW, and SEP in combination with citicoline. Krox-20, NRG-1, and phospholipids expression were measured by immunohistochemical assay, while cognitive function were assessed with the Morris Water Maze test. According to ANOVA test results, it was showed that SEP improved the nerve cells through the activation of Krox-20, NRG-1 expression, and distribution of phospholipids significantly (p <0.05). Based on Morris Water Maze test, SEP also improved the cognitive function in TBI-induced rats. Thus, it can be concluded that spade leaf extract phytosome combined with citicoline increase a higher phospholipids distribution and give the fastest time in the cognitive tests compared with of spade leaf extract phytosome and citicoline alone.Keywords : NRG-1, Krox-20, phospholipids, phytosome, Centella asiatica

Potensi Herbal Delivery System Ekstrak Pegagan sebagai Modulator Neuregulin-1 (NRG-1) pada Tikus Model Traumatic Brain Injury (TBI) Menggunakan Teknologi Phytosome

TBI ialah gangguan fungsi otak akibat benturan yang parah. Secara molekuler, terjadi kerusakan akson yang menyebabkan terjadinya penurunan sintesis NRG-1 yang berperan pada proses remielinasi. Pegagan diketahui memiliki kandungan bahan aktif yang dapat memicu aktivasi NRG-1. Bahan aktif yang bersifat polar membutuhkan modifikasi kepolaran untuk meningkatkan absorbsi dan bioavaibilitas bahan aktif menggunakan teknologi phytosome. Penelitian ini bertujuan membuktikan dan membandingkan efektivitas ekstrak dan phytosome dalam optimalisasi herbal delivery system ekstrak Pegagan sebagai neuroprotektor yang ditandai dengan peningkatan ekspresi NRG-1 pada model tikus TBI. Pegagan diekstraksi menggunakan metode maserasi. Setelah sebagian ekstrak diformulasikan dalam bentuk phytosome, dilakukan karakterisasi menggunakan KLT, LC/MS-MS dan SEM. Induksi TBI menggunakan metode closed-head injury. Tikus sebanyak 32 ekor dikelompokkan menjadi kelompok: kontrol (-), kontrol (+), 3 kelompok diberi ekstrak Pegagan, dan 3 kelompok phytosome ekstrak Pegagan dengan dosis masing-masing 90 mg/kgBB, 180 mg/kgBB, dan 360 mg/kgBB. Pengukuran ekspresi NRG-1 dilakukan dengan cara mengamati slide preparat setelah dilakukan pengecatan imunohistokimia. Hasil uji ANOVA (p = 0,000) menunjukkan adanya pengaruh signifikan terapi terhadap ekspresi NRG-1. Dari hasil Uji post-hoc Tukey didapatkan perbedaan bermakna antara kontrol positif dengan semua kelompok ekstrak dan phytosome, tetapi kelompok phytosome hanya pada dosis 180 mg/kgBB, dan 360 mg/kgBB. Kesimpulan penilitian ini ialah phytosome ekstrak Pegagan dapat meningkatkan ekspresi Neuregulin-1 yang lebih tinggi dibandingkan dengan ekstrak Pegagan saja