NUCB2/Nesfatin-1对滋养层细胞合体化的调节作用及膳食必需脂肪酸的影响

胎盘滋养层细胞分化异常是多种妊娠并发症的病因基础，严重危害母胎健康。基于前期研究，课题组认为NUCB2具有抑制细胞滋养层细胞分化融合的作用，而α-亚麻酸能够抑制其表达而促进细胞滋养层细胞合体化。为此，拟首先建立NUCB2过表达转基因动物，并用α-亚麻酸干预，观察胎盘形态和滋养层细胞的分化融合及胎鼠生长发育；再以BeWo和人胎盘滋养层细胞为对象，用NUCB2的活性形式Nesfatin-1处理，并构建NUCB2抑制/过表达细胞模型，确定NUCB2对滋养层细胞分化融合的作用和机制及α-亚麻酸的影响；最后以胎儿生长受限孕妇为对象，检测胎盘NUCB2的表达及Nesfatin-1和脂肪酸水平，分离滋养层细胞观察其对α-亚麻酸、EPA和DHA的反应；最终阐述NUCB2调节滋养层细胞分化融合的作用和机制及膳食脂肪酸的影响，为理解营养素在胎盘发育及胎盘相关疾病治疗和预防中的作用提供基础数据。

It is one of the essential biological processes that mononuclear cytotrophoblasts fuse and differentiate into the multinucleated syncytiotrophoblast (syncytialization) during the gestation. Syncytialization is a hallmark of placentation and is essential for the maintenance of pregnancy and for fetal growth. Based on the preliminary studies and the references, it is hypothesized that NUCB2 might inhibit this process, but α-linolenic acid could inhibit the expression of NUCB2 and promote the cytotrophoblast cell differentiation. Overexpression of NUCB2 C57 mice will be achieved by injection of adeno-associated virus vectors of NUCB2. Placental morphology, differentiation and fusion of trophoblast cells and growth and development of fetal mice were observed in transgenic and wild type C57 mice with or without α-linolenic acid feeding. .BeWo and human placental trophoblast cells were cultured and treated with Nesfatin-1, the active form of NUCB2 protein, or n-3 fatty acid. NUCB2 inhibition and overexpression BeWo cells will be achieved by transferred with NUCB2 inhibition RNA or full NUCB2 sequence. Cell morphology, hCG, HERVWs and other genes expression will be measured by real-time PCR and Western blot. The PI3K/AKT/mTOR signal pathway will also be measured to show the mechanism of the inhibiting of NUCB2 on the syncytialization. Finally, pregnant women with fetal growth restriction will be recruited, the placenta NUCB2 expression, maternal and child serum Nesfatin-1 and fatty acid levels will be measured. The placenta trophoblast cells of cases and controls will be separated and treated withα-linolenic acid, EPA, DHA. Based on these studies, the role and mechanism of NUCB2 in regulating the differentiation and fusion of cytotrophoblast cells, as well as the influence of dietary fatty acids will be explored, which will help understanding the role of nutrients in placental development and the prevention of placental-related diseases.