Abán C, Martinez N, Carou C, Albamonte I, Toro A, Seyahian A, Franchi A, Leguizamón G, Trigubo D, Damiano A, Farina M
Laboratorio de Fisiopatología Placentaria, Centro de Estudios Farmacológicos y Botánicos (CEFYBO, CONICET), Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, 16th floor, C1121ABG, Buenos Aires, Argentina.
Laboratorio de Biología de la Reproducción, Instituto de Fisiología y Biofísica Bernardo Houssay (IFIBIO) - CONICET- Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina.
Apoptosis. 2016 Oct;21(10):1094-105. doi: 10.1007/s10495-016-1274-x.
During pregnancy, apoptosis is a physiological event critical in the remodeling and aging of the placenta. Increasing evidence has pointed towards the relevance of endocannabinoids (ECs) and hypoxia as modulators of trophoblast cell death. However, the relation between these factors is still unknown. In this report, we evaluated the participation of ECs in placental apoptosis induced by cobalt chloride (CoCl2), a hypoxia mimicking agent that stabilizes the expression of hypoxia inducible factor-1 alpha (HIF-1α). We found that HIF-1α stabilization decreased FAAH mRNA and protein levels, suggesting an increase in ECs tone. Additionally, CoCl2 incubation and Met-AEA treatment reduced cell viability and increased TUNEL-positive staining in syncytiotrophoblast layer. Immunohistochemical analysis demonstrated Bax and Bcl-2 protein expression in the cytoplasm of syncytiotrophoblast. Finally, HIF-1α stabilization produced an increase in Bax/Bcl-2 ratio, activation of caspase 3 and PARP cleavage. All these changes in apoptotic parameters were reversed with AM251, a CB1 antagonist. These results demonstrate that HIF-1α may induce apoptosis in human placenta via intrinsic pathway by a mechanism that involves activation of CB1 receptor suggesting a role of the ECs in this process.
在孕期，细胞凋亡是胎盘重塑和老化过程中的一个关键生理事件。越来越多的证据表明，内源性大麻素（ECs）和缺氧作为滋养层细胞死亡的调节因子具有相关性。然而，这些因素之间的关系仍不清楚。在本报告中，我们评估了ECs在由氯化钴（CoCl2）诱导的胎盘细胞凋亡中的作用，CoCl2是一种缺氧模拟剂，可稳定缺氧诱导因子-1α（HIF-1α）的表达。我们发现HIF-1α的稳定降低了脂肪酸酰胺水解酶（FAAH）的mRNA和蛋白水平，提示ECs水平升高。此外，CoCl2孵育和Met-AEA处理降低了细胞活力，并增加了合体滋养层中TUNEL阳性染色。免疫组织化学分析显示Bax和Bcl-2蛋白在合体滋养层细胞质中表达。最后，HIF-1α的稳定导致Bax/Bcl-2比值增加、半胱天冬酶3激活和聚（ADP-核糖）聚合酶（PARP）裂解。使用CB1拮抗剂AM251可逆转凋亡参数的所有这些变化。这些结果表明，HIF-1α可能通过涉及CB1受体激活的机制，经由内源性途径诱导人胎盘细胞凋亡，提示ECs在这一过程中发挥作用。
