Pengaruh Pemberian Ekstrak Kemangi (Ocimum Sanctum) terhadap Ekspresi dan Distribusi Serotonin (5-HT) pada Model In Vivo dan In Vitro Penyakit Neurodegeneratif

MOCHAMMAD NOVA R, drh. Dwi Liliek Kusindarta, MP., Ph.D.; Dr.med.vet. drh. Hevi Wihadmadyatami, M.Sc.

2020 | Tesis | MAGISTER SAINS VETERINER

Penderita penyakit neurodegeneratif (neurodegenerative diseases, NDD) di seluruh dunia mencapai 50 juta orang. Diperkirakan total penderita penyakit ini akan mencapai 82 juta di tahun 2030 dan 152 juta di tahun 2050. Kemangi (Ocimum sanctum) dikenal sebagai tanaman obat seperti antiinflamasi, antioksidan, antialergi, antikarsinogenik, dan sebagai agen neuroprotektan. Penderita neurodegeneratif mengalami kematian neuron otak disertai penurunan level neurotransmiter termasuk serotonin (5-HT). Serotonin merupakan salah satu neurotransmiter yang berperan dalam kemampuan belajar dan memori serta memodulasi pelepasan neurotransmiter lainnya. Penelitian ini bertujuan untuk mengetahui pengaruh pemberian OSEterhadap distribusi dan ekspresi serotonin (5-HT) secara in vitro pada cell line model NDD dan in vivo pada tikus model NDD setelah induksi Trimethyltin Chloride (TMT). Penelitian secara in vitro menggunakan sel HEK-293 dengan 4 perlakuan berbeda yaitu kontrol (tanpa perlakuan), OSE, TMT, dan OSE + TMT. Dilakukan pembuatan lisat dari sel HEK-293 lalu diukur ekspresi 5-HT menggunakan ELISA kompetitif. Penelitian secara in vivo menggunakan 65 ekor tikus putih Wistar (Rattus norvegicus) jantan yang dibagi 3 kelompok yaitu A (kontrol), B (OSE 100 mg/kg BB pre-induksi, induksi TMT 8 mg/kg BB), C (OSE 100 mg/kg BB pre- dan pascainduksi, induksi TMT 8 mg/kg BB). Setiap minggu hingga 21 hari pasca induksi tikus dikorbankan untuk diambil otaknya dilanjutkan dengan pewarnaan cressyl violet dan imunohistokimia terhadap 5-HT. Data pewarnaan dianalisis secara semi kuantitatif untuk menghitung kepadatan neuron dan ekspresi 5-HT pada tikus model NDD. Level 5-HT sel HEK-293 terlihat turun secara signifkan pada perlakuan TMT dibandingkan kontrol. Sedangkan pada perlakuan OSE dan OSE + TMT 5-HT berhasil dipertahankan pada level normal seperti pada kontrol. Gambaran histologi kelompok B menunjukkan penurunan rata-rata jumlah neuron dan sel imunoreaktif 5-HT pada area hipokampus hingga hari ke-21 pasca-induksi TMT. Gambaran histologi kelompok C menunjukkan peningkatan signifikan rata-rata jumlah neuron dan sel imunoreaktif 5-HThipokampus hingga hari ke-21 pasca-induksi TMT dibanding kelompok B (Tabel 2) (A vs B vs C = ++++ vs ++ vs +++). OSEmemiliki aktivitas neuroprotektif bahkan neuroregeneratif terbukti dengan menjaga 5-HT sel HEK-293 pada level normal saat diinduksi TMT serta melalui peningkatkan kepadatan neuron dan ekspresi 5-HT pada hipokampus model NDD yang diinduksi TMT.

Patients with neurodegenerative diseases (NDD) around the world reach 50 million people. It is estimated that the total sufferers of this disease will reach 82 million in 2030 and 152 million in 2050. Basil (Ocimum sanctum) is known as a medicinal plant that has many actions such as anti-inflammatory, antioxidant, antiallergic, anticarcinogenic, and as a neuroprotectant agent. Neurodegenerative patients severe neuronal death in brain accompanied by decreased levels of neurotransmitters including serotonin (5-HT). Serotonin is a neurotransmitter that plays a role in learning and memory and modulates the release of other neurotransmitters. This study aims to determine the effect of OSE administration on serotonin (5-HT) distribution and expression in vitro on NDD and in vivo cell line models in NDD model mice after induction of trimethyltin chloride (TMT). In vitro study used HEK-293 cells with 4 different treatments namely control (without treatment), OSE, TMT, and OSE + TMT. Making lysate from HEK-293 cells was then measured 5-HT expression using competitive ELISA. In vivo study used 65 male Wistar (Rattus norvegicus) white rats divided into 3 groups namely A (control), B (OSE 100 mg / kg pre-induction BW, TMT induction 8 mg / kg BW), C (OSE 100 mg / kg BW pre- and postduction, TMT induction 8 mg / kg BW). Every week until 21 days after induction of rats sacrificed to be taken the brain then performed cressyl violet staining and immunohistochemistry against 5-HT. Staining data were analyzed semi-quantitatively to calculate neuron density and 5-HT expression in NDD model mice. Serotonin level of HEK-293 was seen to decrease significantly in TMT treatment compared to controls. Whereas in the treatment of OSE and OSE + TMT 5-HT successfully maintained at normal levels as in the control. Histological features of group B showed a decrease in the average number of neurons and 5-HT immunoreactive cells in the hippocampal area until the 21st day post-induced TMT. Histological features of group C showed a significant increase in the average number of hippocampal 5-HT neuroreactive and immunoreactive cells up to the 21st day post-induced TMT compared to group BB (Table 2) (A vs B vs C = ++++ vs ++ vs +++). OSE has neuroprotective and even neuroregenerative activity proven by maintaining 5-HT HEK-293 at normal levels against TMT induced and through increasing neuron density and 5-HT expression in the hippocampus of TMT-induced NDD models.

Kata Kunci : neurodegeneratif, OSE, hipokampus, serotonin, sel HEK-293