Regulatory Effects of Thymoquinone on Dopamine Level in Neuronal Cells Exposed to Amphetamine: An In Vitro Study

Introduction: Amphetamine (AT) is used to treat some medical conditions and also known to be abused recreationally. It is a potent central nervous system stimulant that is capable of producing damaging effects to the central dopaminergic pathway. Most of AT users are treated clinically for symptomatic treatment which is associated with neurological side effects. To date, there is growing interest in naturally occurring compounds which have lesser side effects to treat health problems. One of the potential compounds is thymoquinone (TQ), an active compound of Nigella sativa which is known for its cellular protective effects. Objective: The objectives of this study were to determine the IC50 values of AT and TQ on differentiated SH-SY5Y neuronal cells and to evaluate the changes of dopamine (DA) level in the cells exposed to AT after co-administering with TQ. Methodology: Differentiated SH-SY5Y cells were grown in cell culture flask containing DMEM/F12 medium supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) penicillin/streptomycin. The IC50 value of TQ and AT in differentiated SH-SY5Y cells was determined by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The DA level was determined by using the Enzyme-Linked Immunosorbent Assay (ELISA) kit. Result and Discussion: The IC50 values of AT and TQ were 1596 µM and 926 µM respectively. Co-administration of 40 µM of AT and 30 µM of TQ demonstrated a significant increase in DA level at 48 hours of exposure when compared to the administration of AT group (P≤0.05). Conclusion: These findings suggested that TQ has a role in maintaining the DA activity after a long-term AT exposure

MDMA-induced BV2 microglial cell activation in vitro

Background: 3,4-Methylenedioxymethamphetamine (MDMA) is a psychostimulant drug that induces neurotoxicity. Even though several psychostimulant substances activate microglia, little is known about MDMA's effects on these cells, and evidence of MDMA-induced microglial activation is equivocal. Materials and Methods: This study employed a murine microglial cell line, BV2, to examine the effects of MDMA on the microglia morphological changes and the survival of microglia in vitro. MDMA was incorporated into the media at the time of plating, and cell number and mitochondrial dehydrogenase activity (MTT) levels were determined in vitro. The level of pro-inflammatory cytokine TNF-α was also determined. Results: Treatment of BV2 cells with MDMA resulted in morphological changes, reduced cell viability after 24h incubation with the inhibitory concentration (IC50) value of 243.6 µg/mL, and increased TNF-α level in a dose-dependent manner. Conclusion: These findings proposed that MDMA could induce BV2 microglial cell activation in vitro and suggested that it has an essential role in developing MDMA use disorder

EFFECTS OF ZAMZAM WATER AND METHADONE ON THE EXPRESSION OF MU-OPIOID RECEPTOR-1 GENE IN MORPHINE-DEPENDENT RATS AFTER CHRONIC MORPHINE ADMINISTRATION

Background: Sodium ion is an essential ion that is implicated in many physiological functions. Recently, sodium ion was reported to facilitate the activation of Mu-Opioid Receptor (MOR) by binding at the allosteric site of the MOR. Zamzam water is water originated from Mecca. Couple of studies proved that Zamzam water has the therapeutic effect owing to its mineral. In this study, we want to determine the ion concentration of Zamzam water and then to investigate the effects of Zamzam water and co-treatment with methadone on the regulation of MOR-1gene after chronic morphine administration. Materials and Methods: Zamzam water, tap water and normal mineral water were analyzed using Ion chromatography. Meanwhile, in animal study, 50 male Sprague Dawley rats were randomly divided into five groups. All group of rat were made dependence on morphine using intraperitoneal injection except for normal group. Morphine dependent rats then were treated with methadone, Zamzam water and co-treatment methadone with Zamzam water for thirty days, respectively. The Ventral Tegmental Area (VTA) of rat’s brain was dissected and subjected to real-time quantitative RT-PCR to determine the regulation of MOR-1 gene expression. The obtained data were analyzed using SPSS v.11 software, and one-way ANOVA followed by Tukey’s Post-test. Results: The data obtained showed that Zamzam water is significantly high in ion concentration compared to tap water and normal mineral water. Besides, the result from gene expression analysis showed co-treatment Zamzam water and methadone significantly prevented the downregulation of MOR as compared to methadone and Zamzam water treatment alone (

Effects of apigenin, luteolin, and quercetin on the natural killer (NK-92) cells proliferation: a potential role as immunomodulatory

Cancer can be classified as a fourth leading cause of death in Malaysia. There is a continuous effort by scientists in finding alternative cure to cancer due to the known side effects of chemotherapy and radiation therapy as well as recurrences. One of the latest methods to kill cancerous cells is by using immune cells known as natural killer (NK) cells. Flavonoids such as flavone and flavonol are also known for their antioxidant, anti-inflammatory, immunomodulatory and anticancer properties. This study was carried out to determine the role of flavonoid compounds of apigenin, luteolin, and quercetin to facilitate the growth of NK-92 cells. NK-92 cell line was grown in tissue culture flasks containing α- Minimum Essential Medium (MEM) medium enriched with L-glutamine, 12.5% fetal bovine serum, 12.5% horse serum, 0.2 mM myo-inositol, 0.02 mM folic acid, and 100 - 200 U/mL recombinant interleukin 2 (IL-2). The cell viability was determined via trypan blue staining where the cells were manually counted by a haemocytometer. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cell viability of NK-92 cells after treatment with apigenin, luteolin, and quercetin. Results showed a dose-dependent proliferative effects of apigenin, luteolin, and quercetin on the proliferation of NK-92 cells showing the highest percentage of proliferation at 100 µg/mL for all compounds (*P < 0.05). However, exceeding the dose of 100 µg/mL had resulted in a decline of cell proliferations percentage. Based on these findings, flavonoid compounds comprising apigenin, luteolin and quercetin were able to induce proliferative effects on NK-92 cells