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Zinc Supplementation Reduces ROS Production and Prevents MDMA-Induced Apoptosis in TM3 Leydig Cells via the
Salar Mahmoudi-Nejad1, Sina Ahmadi1, Mahssa Hassan-Nejhad1
1Student Research Committee, Urmia University of Medical Sciences, Urmia, Iran.
Abstract:
MDMA can cause serious adverse effects on vital organs such as the heart, brain, and liver. Additionally, MDMA consumption can also potentially cause various endocrine system dysfunctions. The previous study has shown that pre-treatment of zinc can reduce the cytotoxicity of MDMA on the Leydig cell line (TM3). In this study, we investigated the mechanisms involved in the treatment with MDMA on the apoptosis of TM3 cells and the effects of zinc pre-treatment on reducing the apoptotic effects of MDMA. TM3 cells were incubated with MDMA (5 mM), zinc (8 µM), and zinc (8 µM) prior to MDMA (5 mM) for 48 h. The cells were pre-treated with zinc for 24 h prior to the administration of MDMA, and the total culture time was 48h. The effect of different treatment groups in causing oxidative stress and apoptosis in TM3 cells was measured by DCF, TUNNEL, and western blot tests, respectively. Our results revealed that the number of DCF and tunnel-positive cells increases as a result of MDMA treatment. In addition, the treatment with MDMA increased the expression of pro-apoptotic proteins caspase 3, Bax, and p53. Conversely, the expression of anti-apoptotic protein Bcl-2 decreased. Zinc pre-treatment significantly decreased the expression of pro-apoptotic proteins and the number of tunnels and DCF-positive cells compared to the MDMA-only group. It is concluded that MDMA has a toxic effect and causes apoptosis on TM3 cells, and also, pre-treatment with zinc mitigates the ROS production and toxic effect of MDMA and MDMA-induced apoptosis in TM3 cells.
Insights
3,4-Methylenedioxymethamphetamine (MDMA) induces apoptosis in Leydig cells, causing oxidative stress. Zinc pre-treatment mitigates these toxic effects by reducing reactive oxygen species and apoptosis markers.
Area of Science:
- Endocrinology
- Toxicology
- Cell Biology
Background:
- 3,4-Methylenedioxymethamphetamine (MDMA) is known to cause adverse effects on vital organs and endocrine functions.
- Previous research indicates zinc may reduce MDMA's cytotoxicity in Leydig cells (TM3).
Purpose of the Study:
- To investigate the mechanisms of MDMA-induced apoptosis in TM3 cells.
- To evaluate the protective effects of zinc pre-treatment against MDMA toxicity.
Main Methods:
- TM3 cells were treated with MDMA, zinc, or zinc followed by MDMA.
- Oxidative stress and apoptosis were assessed using DCF, TUNNEL assays, and Western blotting for apoptosis-related proteins.
Main Results:
- MDMA treatment increased oxidative stress (DCF-positive cells) and apoptosis (TUNNEL-positive cells) in TM3 cells.
- MDMA upregulated pro-apoptotic proteins (caspase 3, Bax, p53) and downregulated anti-apoptotic protein (Bcl-2).
- Zinc pre-treatment significantly reduced MDMA-induced oxidative stress and apoptosis markers.
Conclusions:
- MDMA exhibits toxic effects, inducing apoptosis and oxidative stress in TM3 Leydig cells.
- Zinc pre-treatment effectively mitigates MDMA's toxic effects, reducing reactive oxygen species production and apoptosis.
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