Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, Center for DAMP Biology, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510510, People's Republic of China.
Department of Neurology, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510510, People's Republic of China.
Neurotox Res. 2020 Dec;38(4):859-870. doi: 10.1007/s12640-020-00216-x. Epub 2020 May 16.
Duloxetine is a clinical drug that is primarily used for treatment of depression and pain, but it has side effects of addiction and tolerance. Cytochrome P450 (CYP) is its metabolic enzyme, and the drug's biofunction results from its neuro-protective effect in animal and cell models. We aimed to investigate the duloxetine-induced neural cytotoxicity effect and its performance in an N2a cell neurite outgrowth model. Cell death was assessed as cell viability using a Cell Count Kit-8 and further evaluated using bright-field images, propidium iodide (PI) and annexin V staining, colony-formation analysis, TUNEL staining of the cells, and biochemical testing. N2a cells were committed to differentiation by serum withdrawal and RA induction, and the neurite outgrowth was evaluated as the number of differentiated cells, longest neurite length, and average neurite length. Cell cycle analysis, PI and annexin V staining, mRNA expression, and biochemical testing were used to evaluate the drug effects on differentiation. The induction of neural cell death by duloxetine was not affected by classic cell death inhibitors but was promoted by the CYP inducer rifampicin. N2a cell neurite outgrowth was promoted by duloxetine via reduction of the CYP2D6 and MDA levels and induction of Bdnf protein levels. Duloxetine induces neural cell death through effects on CYP and promotes N2a cell neurite outgrowth by regulating CYP, Bdnf protein, and the intracellular lipid peroxidation level.
度洛西汀是一种临床药物，主要用于治疗抑郁症和疼痛，但它有成瘾和耐受的副作用。细胞色素 P450（CYP）是其代谢酶，药物的生物功能来自于其在动物和细胞模型中的神经保护作用。我们旨在研究度洛西汀诱导的神经细胞毒性作用及其在 N2a 细胞突起生长模型中的表现。细胞死亡通过 Cell Count Kit-8 作为细胞活力进行评估，并通过明场图像、碘化丙啶（PI）和 Annexin V 染色、集落形成分析、细胞 TUNEL 染色和生化测试进一步评估。N2a 细胞通过血清撤离和 RA 诱导进行分化，突起生长作为分化细胞的数量、最长突起长度和平均突起长度进行评估。细胞周期分析、PI 和 Annexin V 染色、mRNA 表达和生化测试用于评估药物对分化的影响。度洛西汀诱导的神经细胞死亡不受经典细胞死亡抑制剂的影响，但受 CYP 诱导剂利福平的促进。度洛西汀通过降低 CYP2D6 和 MDA 水平和诱导 Bdnf 蛋白水平促进 N2a 细胞突起生长。度洛西汀通过影响 CYP 诱导神经细胞死亡，并通过调节 CYP、Bdnf 蛋白和细胞内脂质过氧化水平促进 N2a 细胞突起生长。
