PENGARUH PEMBERIAN ECTASY (METHYLENEDIOXYMETHAMPHETAMINE) TERHADAP POLA KEMATIAN SEL SYARAF MENCIT MUS MUSCULLUS SPECIES INVITRO (The effects of ecstacy (methylenedioxytnethamphetamine)on the mus muscullus species neuronal death types invitro study)

Abstract

Background 3,4-Methylenedioxymethamphetamine (MDMA, "Ecstasy") is a derivative of amphetamine that has become extensively used as a recreational drug. Although MDMA has been popularly considered as a safe drug, there are increasing evidences of its toxicity. MDMA is neurotoxic for serotonergic and dopaminergic neurons and produces a decrease of 5-HT transporters. Other side-effects of MDMA are hyperthermia, rhabdomyolysis and cardiac dysrhythmias. Recent publication data shows that the neurotoxic effects of MDMA is increasing by the degree of an animal model. Neuron to be affected will undergoing degeneration and finally to be death. When the cell is exposed to physical, biochemical or biological injury, or deprived of necessary substances, it activates a series of stress-response genes. With minimal insults, the cell may recover. With greater insults, single cell death, or apoptosis, results; the cell dies and is recycled to its neighbours. If the insult overwhelms a large number of cells then necrosis ensues, with an accompanying inflammatory response. Otherwise, there was lack of publication data according to the effects of MDMA neorutoxicity, especially types of neuronal cell death in vitro study. Objective : To establish effects of MDMA dose and time duration according to apoptosis and necrosis of the nerve cell in vitro study. Methode : The Study using Simple Randomize Subject Design. Tissue culture was done according to the common methode. MDMA treatment was conducting to the optimized culture (cell count 1061m1). Control group only get Nacl 0,9%. Group I under 0,250 mg of MDMA. Group II under 0,500 mg of MDMA. Group III under 0,750 mg of MDMA. Group IV under 1,000 mg of MDMA. Group V under 1,250 mg of MDMA. Dualstaining with Acridine Orange (AO) dan Propidium Iodide (PI) were done to differentiated of apoptosis and necrosis cells. We were conducting examination in four time series (i.e 4 hours, 12 hours, 24 hours, and 48 hours). Mean difference among the group were calculated using one-way anova. To determined power and direction of the independence variable (dose and time duration of MDMA exposure) influences the dependence variable (percentage apoptosis or necrosis of the cell) using linear regression analysis. Data analysis was conducted using SPSS version 11.01 for Widows under 0,05 significance level. Result : There were significant difference of the mean percentage of apoptotic cells among the groups of MDMA dose.( p=0,004) The increasing dose of MDMA is correlated to the increasing percentage of apoptotic nerve cells in vitro study.(r=0,339) There were significant difference of the mean percentage of apoptotic cells among the groups of duration time of MDMA exposure.( p=0,000) The increasing duration time of MDMA exposure is correlated to the increasing percentage of apoptotic nerve cells in vitro study.(r=0,710) There were significant difference of the mean percentage of necrotic cells among the groups of MDMA dose.( p=0,005) The increasing dose of MDMA is correlated to the increasing percentage of necrotic nerve cells in vitro study.(r=0,329) There were significant difference of the mean percentage of necrotic cells among the groups of duration time of MDMA exposure.( p=0,000) The increasing duration time of MDMA exposure is correlated to the increasing percentage of necrotic nerve cells in vitro study.(r=0,769) Conclusion : Statistically, there were correlation between dose and time durations of MDMA exposure to the percentage of apoptotic and necrotic nerve cell invitro study Latar Belakang : Penyalahgunaan Ecstacy atau Methylendioxymethamphetamine (MDMA) cenderung meningkat walaupun telah didapatkan cukup bukti adanya pengaruh buruk terhadap berbagai organ (jantung, hati, ginjal, dan otak). Telah banyak publikasi tentang sifat neurotoksisitas MDMA yang dilakukan pada berbagai hewan percobaan. Makin tinggi derajad hewan percobaan sifat neurotoksiknya makin tinggi. Sifat neurotoksik MDMA terutama terhadap syaraf serotonergik dan dopaminergik. Neuron yang terkena akan mengalami degenerasi dan berakhir dengan kematian. Secara umum kematian sel terjadi melalui nekrosis atau apoptosis. Belum banyak penelitian yang dilakukan untuk membuktikan kejadian apoptosis atau nekrosis sel syaraf akibat pengaruh ecstacy. Penelitian ini dilakukan invitro dengan alasan agar dapat mengendalikan factor-faktor pengganggu sehingga diharapkan efek yang timbul benar¬benar hanya disebabkan pengaruh pemaparan MDMA. Tujuan Penelitian : Membuktikan adanya pengaruh dosis dan lama waktu pemaparan MDMA terhadap apoptosis dan nekrosis sel syaraf invitro. Metode Penelitian : Penelitian dilakukan secara invitro dengan menumbuhkan sel syaraf pada suatu tabung (flask). Setelah kultur mencapai tingkat pertumbuhan yang optimun selanjutnya diberi perlakukan dengan pemberian MDMA dengan dosis tertentu sesuai dengan kelompok penelitian. Kelompok kontrol hanya mendapatkan cairan fisiologis, Kelompok I mendapatkan dosis 0,250 mg, Kelompok II mendapatkan dosis 0,500 mg, Kelompok III mendapatkan 0,750 mg, Kelompok N mendapatkan 1,000 mg, dan Kelompok V mendapatkan 1,025 mg. Pemeriksaan adanya apoptosis dan nekrosia dilakukan dengan pengecatan dual staining dengan Acridine Orange (AO) danPropidium Iodide (PI). Pemeriksaan pada masing-masing kelompok dilakukan sebanyak 4 kali yaitu pada jam ke-6, 12, 24 dan 48. Untuk mengetahui perbedaan rerata variable tergantung (persentase sel apoptosis atau persentase sel nekrosis) pada masing-masing kelompok dilakukan dengan uji statistik analisis varian satu arah (one-way anova). Untuk mengetahui kekuatan dan arah hubungan antara variable tergantung (persentase sel apoptosis atau persentase sel nekrosis) dengan variable bebas (dosis dan lama waktu pemaparan MDMA) 'dilakukan dengan analisis regresi linear. Analisis data dilakukan dengan program SPSS 11.01 for Window's. Batas signifikansi a=0,05. Hasil Penelitian : Terdapat perbedaan bermakna rerata persentase apoptosis sel syaraf pada berbagai kelompok dosis MDMA.(p=0,004) Peningkatan dosis MDMA meningkatkan persentase apoptosis sel syaraf invitro. (=0,339) Terdapat perbedaan bermakna rerata persentase apoptosis sel syaraf pada berbagai kelompok lama waktu pemaparan MDMA.(p=0,000) Peningkatan lama waktu pemaparan MDMA meningkatkan persentase apoptosis sel syaraf invitro. (r=0,710) Terdapat perbedaan bermakna rerata persentase nekrosis sel syaraf pada berbagai kelompok dosis MDMA. (p=0,005) Peningkatan dosis MDMA meningkatkan persentase nekrosis sel syaraf invitro.(r=0,329) Terdapat perbedaan bermakna rerata persentase nekrosis sel syaraf pada berbagai kelompok lama waktu pemaparan MDMA.(p=0,000) Peningkatan lama waktu pemaparan MDMA meningkatkan persentase nekrosis sel syaraf invitro. (r--0,769). Simpulan : Secara statistik terdapat hubungan antara dosis dan lama waktu pemaparan MDMA dengan persentase apoptosis maupun nekrosis sel syaraf invitro.