Lee Shih-Yu, Chiu Shang-Wen, Li I-Hsun, Tsai Wei-Cheng, Li Chung-Hsing
Graduate Institute of Aerospace and Undersea Medicine, National Defense Medical Center, Taipei, Taiwan.
Division of Orthodontics, Pediatric Dentistry, and Special Needs Dentistry, Department of Dentistry, Tri-Service General Hospital, Taipei, Taiwan.
J Dent Sci. 2025 Jul;20(3):1782-1791. doi: 10.1016/j.jds.2025.04.025. Epub 2025 May 6.
BACKGROUND/PURPOSE: 3,4-methylenedioxymethamphetamine (MDMA) is a synthetic substituted amphetamine. Research primarily focuses on its neurotoxicity and psychological effects, while studies examining its impact on mesenchymal stem cells (MSCs) and dental pulp stem cells (DPSCs) are relatively limited. This study investigated the cytotoxicity and molecular mechanisms of MDMA in DPSCs.
Cell viability, apoptosis, autophagy, immunophenotype, reactive oxygen species (ROS), and the related signaling pathways were analyzed using a cell counting kit, Western blot, flow cytometry, and 2',7'-dichlorofluorescein diacetate (DCFH-DA) dye after treating DPSCs with indicated concentrations of MDMA.
MDMA significantly decreased cell viability and increased cleaved poly adenosine diphosphate-ribose polymerase (PARP), cleaved caspase-8, cleaved caspase-3, and Bcl-2-associated X protein (Bax), while reducing B-cell lymphoma 2 (Bcl-2). Annexin V and 7-aminoactinomycin D (7-AAD) flow cytometry showed that these cells underwent early apoptosis without altering MSCs' immunophenotypic properties. MDMA also induced ROS accumulation and nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) inflammasome activation. Autophagy activation was observed with adenosine monophosphate-activated protein kinase (AMPK) activation, protein kinase B (AKT) inactivation, and mammalian target of rapamycin (mTOR) suppression. Autophagy inhibitor chloroquine (CQ) attenuated MDMA-induced cell death, suggesting that excessive autophagy contributes to cell damage.
MDMA induces ROS-mediated autophagic cell death and thioredoxin-interacting protein (TXNIP)/NLRP3 inflammasome activation, causing detrimental cell damage. The historical roots of drug addiction should be considered when modifying dental approaches for patients with a history of MDMA use, as well as in the screening of DPSCs donors and patients undergoing stem cell therapy.
背景/目的：3,4-亚甲基二氧甲基苯丙胺（摇头丸）是一种合成的取代苯丙胺。研究主要集中在其神经毒性和心理影响，而研究其对间充质干细胞（MSC）和牙髓干细胞（DPSC）影响的相对较少。本研究调查了摇头丸对DPSC的细胞毒性和分子机制。
用指定浓度的摇头丸处理DPSC后，使用细胞计数试剂盒、蛋白质免疫印迹法、流式细胞术和2',7'-二氯二氢荧光素二乙酸酯（DCFH-DA）染料分析细胞活力、凋亡、自噬、免疫表型、活性氧（ROS）及相关信号通路。
摇头丸显著降低细胞活力，增加裂解的聚腺苷二磷酸核糖聚合酶（PARP）、裂解的半胱天冬酶-8、裂解的半胱天冬酶-3和Bcl-2相关X蛋白（Bax），同时降低B细胞淋巴瘤-2（Bcl-2）。膜联蛋白V和7-氨基放线菌素D（7-AAD）流式细胞术显示这些细胞发生早期凋亡，而不改变MSC的免疫表型特性。摇头丸还诱导ROS积累和核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）炎性小体激活。观察到自噬激活伴有腺苷单磷酸活化蛋白激酶（AMPK）激活、蛋白激酶B（AKT）失活和雷帕霉素靶蛋白（mTOR）抑制。自噬抑制剂氯喹（CQ）减弱了摇头丸诱导的细胞死亡，表明过度自噬导致细胞损伤。
摇头丸诱导ROS介导的自噬性细胞死亡和硫氧还蛋白相互作用蛋白（TXNIP）/NLRP3炎性小体激活，导致有害的细胞损伤。在修改对有摇头丸使用史患者的牙科治疗方法时，以及在筛选DPSC供体和接受干细胞治疗的患者时应考虑药物成瘾的历史根源。
