n-3多不饱和磷脂酰丝氨酸对大鼠海马神经元细胞的抗凋亡作用研究

Study on the anti-apoptotic effect of n-3 polyunsaturated phosphatidylserine on rat hippocampal neuron cells

中文关键词:  n-3多不饱和磷脂酰丝氨  海马神经元细胞  抗凋亡

英文关键词:n-3 polyunsaturated phosphatidylserine  hippocampal neurons cells  anti-apoptotic

中文摘要:

      目的 研究n-3多不饱和磷脂酰丝氨酸对大鼠海马神经元细胞(rat hippocampal neurons cells, RHNC)抗凋亡能力的影响。方法 以RHNC为对象, 以乙酸佛波酯(phorbol 12-myristate 13-acetate, PMA)为诱导剂, 建立凋亡模型。以二十二碳六烯酸(docosahexaenoic acid, DHA)/二十碳五烯酸(eicosapentaenoic acid, EPA)型磷脂酰丝氨酸(DHA/EPA-phosphatidylserine, DHA/EPA-PS)为保护剂, 用吖啶橙染色后在激光共聚焦显微镜下观察细胞不同凋亡时期的形态。通过磷脂酰丝氨酸外翻实验(Annexin V-FITC/PI双染实验), 在细胞水平研究以DHA/EPA-PS为代表的n-3多不饱和磷脂酰丝氨酸对RHNC的抗凋亡能力。结果 在PMA质量浓度为10.0 mg/L、作用时间24 h条件下, 建立RHNC凋亡模型。15.0 mg/L DHA/EPA-PS对RHNC的预保护效果最好, 此时早期、中期凋亡细胞较少且无晚期凋亡细胞, 凋亡率为11.46%±4.11%, 细胞形态和核膜完整, 突起不受PMA的影响。结论 以DHA/EPA-PS为代表的n-3多不饱和磷脂酰丝氨酸对RHNC具有一定的抗凋亡作用。本研究可为磷脂酰丝氨酸的生物活性研究提供较好的数据支撑。

英文摘要:

      Objective To study the effect of n-3 polyunsaturated phosphatidylserine on the anti-apoptotic ability of rat hippocampal neuron cells (RHNC). Methods An apoptosis model using RHNC as the target and phorbol acetate (PMA) as the inducer was established. Using docosahexaenoic acid (DHA)/eicosapentaenoic acid (EPA) phosphatidylserine (DHA/EPA-PS) as a protective agent, the morphology of cells at different stages of apoptosis was observed under a laser confocal microscope after staining with acridine orange. The effect of n-3 polyunsaturated phosphatidylserine, represented by DHA/EPA-PS, on the anti-apoptotic ability of RHNC was studied at the cellular level through the Annexin V-FITC/PI double staining experiment. Results An RHNC apoptosis model with a PMA concentration of 10.0 mg/L and an action time of 24 hours was established. The 15.0 mg/L DHA/EPA-PS showed the best cell pre protection effect, with an apoptosis rate of 11.46%±4.11%. Early and mid-stage apoptotic cells were fewer and did not produce late stage apoptotic cells. Cells pre protected with 15 mg/L DHA/EPA-PS exhibited intact morphology and nuclear membrane, and their protrusions were not affected by PMA. Conclusion n-3 polyunsaturated phosphatidylserine, represented by DHA/EPA-PS, has certain anti-apoptotic ability against RHNC. The study provides good data support for future research on the biological activity of phosphatidylserine.