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目的探讨褪黑素(melatonin,Mel)对巨核细胞凋亡的影响及其作用机制。方法巨核细胞细胞株CHRF-288-11去血清诱导凋亡,加或不加Mel共培养后,流式细胞仪检测凋亡指标:Annexin V/PI、Caspase-3和JC-1,并与正常组和TPO组比较。同时检测信号通路AKT、ERK1/2,了解其参与保护作用的机制。结果Mel 200 nmol/L作用72 h后,CHRF细胞Annexin V/PI、Caspase-3和JC-1的表达较对照组有明显下降,分别由42.9%、29.5%、47.7%降至31.7%(P<0.05)、21.8%(P<0.05)和37.2%(P<0.05)。其中,JC-1的表达与TPO组(20.7±5.2)%比较,差异有统计学意义(P<0.05),而Annexin V/PI、Caspase-3的表达与TPO组比较,差异无统计学意义(P>0.05)。加入Mel后CHRF细胞的磷酸化AKT、ERK1/2水平明显增高,分别由(5.9±0.1)%、(6.1±0.4)%升至(9.5±0.1)%(P<0.05)、(9.3±0.5)%(P<0.05)。结论Mel可抑制巨核细胞凋亡,其保护机制可能通过AKT、ERK信号通路发挥作用。
Objective To investigate the effect of melatonin (Mel) on megakaryocyte apoptosis and its mechanism. Methods The apoptosis of human megakaryocytic cell line CHRF-288-11 was induced by serum. The apoptosis index was measured by flow cytometry (Annexin V / PI, Caspase-3 and JC-1) with or without co-culture with Mel Group and TPO group comparison. At the same time, we detected the signal pathways AKT and ERK1 / 2 to understand the mechanism involved in the protection. Results The expression of Annexin V / PI, Caspase-3 and JC-1 in CHRF cells decreased significantly from 42.9%, 29.5% and 47.7%, respectively, after treated with 200 nmol / L Mel <0.05), 21.8% (P <0.05) and 37.2% (P <0.05) respectively. Among them, the expression of JC-1 was significantly different from that of TPO group (20.7 ± 5.2)% (P <0.05), while the expression of Annexin V / PI and Caspase-3 was not statistically different (P> 0.05). The phosphorylated AKT and ERK1 / 2 levels of CHRF cells increased significantly from (5.9 ± 0.1)% and (6.1 ± 0.4)% to (9.5 ± 0.1)% and (9.3 ± 0.5 )% (P <0.05). Conclusion Mel can inhibit the apoptosis of megakaryocytes, and its protective mechanism may play a role through the AKT and ERK signaling pathways.