铁负荷过重对巨核细胞的凋亡损害及松果体素的保护机制探讨

Protective effect of melatonin on iron-induced apoptosis in CHRF cells

目的探讨铁负荷过重对巨核细胞的凋亡损害及松果体素(Melatonin)的保护机制。方法流式细胞仪检测凋亡指标:Annexin V/PI,Caspase-3和JC-1,同时检测信号通路AKT、ERK1/2,了解铁对巨核细胞株CHRF-288-11的损害机理及-Melatonin参与保护作用的机制。结果0.3mmol/L的氯化铁作用8h引起CHRF细胞明显凋亡,Melatonin200nmol/L可以保护细胞,减少细胞凋亡,Annexin V/PI、Caspase-3和JC-1的表达,分别由37.9%、26.5%、35.7%降至26.9%(P<0.05)、19.0%(P<0.05)和26.0%(P<0.05)。加入Melatonin后细胞磷酸化的AKT、ERK1/2水平明显增高,分别由5.9%、6.1%升至9.5%(P<0.05)、9.3%(P<0.05)。结论Melatonin可能通过激活AKT、ERK信号通路,抑制铁诱导的巨核细胞凋亡,具有保护作用。

Objective To study the effect of iron on megakaryocyte apoptosis and the anti-apoptotic effect of melatonin. Methods Cells of human megakaryocytic cell line CHRF-288-11 were divided into four groups： control （no FeCl3 or melatonin）, melatonin treatment, FCCl3 treatment and FeCl3 plus melatonin treatment. Apoptotic cells were examined by Annexin V-FITC/PI、 Caspase 3-FITC and JC-1 assays using flowcytometry. To study the anti-apoptotic mechanism of melatonin, phospbo-Akt and ERK1/2 were examined by flowcytometry with monoclonal antibodies. Results FeCl3（0.3 mmol/L） induced apoptosis in CHRF cells. Melatonin （200 nmol/L） significantly protected against apoptosis of thesse cells. The population of Annexin V, Caspase 3-and JC-1 were significultly decreased from 37.9%, 26.5%, 35.7% to 26.9% （n=4,P〈0.05）,19.0%（n=4,P〈0.05） and 26.0%（n=4,P〈0.05） respectively. The population of phospho-AKT、 ERK1/2 were significantly increased from 5.9%, 6.1% to 9.5%（n=4,P〈0.05）and 9.3%（n=4,P〈0.05） respectively after treatment by melatonin. Conclusions FeCl3 induces megakaryocyte apoptosis, and this apoptotic effect could be inhibited by melatonin via the activation of AKT and ERK1/2 pathways.