中文摘要：

ER－α36是一种分子量为36kDa的雌激素受体（estrogen receptor,ER）新亚型，其主要通过介导膜雌激素信号通路参与多种细胞生理、病理等过程。本文利用RNA干扰技术敲低大鼠肾上腺嗜铬细胞瘤细胞株PCI2的ER-α36基因，研究ER-α36与Akt在神经细胞缺糖损伤中的关系。采用MTT、Westernblot和AnnexinV／PI双染等方法，观察了ER—α36在细胞缺糖应激反应中的作用。结果显示：（1）随着缺糖损伤时间的增加，PCI2细胞的存活率逐渐降低，6h后其细胞存活率与对照组相比具有极显著性差异（P〈0．01），凋亡率升高，6h后具有极显著性差异（P〈0．01）；葡萄糖转运蛋白Glut-4表达水平显著降低（P〈0．01）；（2）缺糖损伤3h时，ER-α36明显减少（P〈0．01），但随后逐渐增加；缺糖可引起抗凋亡蛋白Akt磷酸化水平先升高后降低（P〈0．05）；加入P13K抑制剂LY294002，ER—α36表达降低，Akt的激活被抑制；（3）敲低ER—α36后缺糖处理，细胞凋亡率高于野生型，具有极显著性差异（P〈0．01）；敲低ER—α36的细胞缺糖导致的Akt激活减弱，Caspase-3表达增多。以上结果表明，在PCI2细胞的缺糖应激反应中，Akt激活情况可能与ER-α36相关，两者共同参与缺糖损伤应激调控。

英文摘要：

ER-α36 is a novel 36-kDa variant of ER-α. A large of evidence demonstrated that ER-α36 responded to membrane-initiated estrogen signaling pathways which were involved in the physiological and pathological process in many kinds of cells. In this study, knock-down of ER-α36 expression in pheochromocytoma （PC12） cells （named as PC12-36L cells） by using the shRNA method was used to evaluate the relationship between ER-α36 and Akt in neurons under glucose deprivation. The effect of ER-α36 on outgrowth of PC 12 cells, as well as the neuroprotective effect of ER-α36 on injured PC 12 cells exposed to glucose deprivation was observed by us- ing MTT assay, Western blot and Annexin V/PI staining et al. The results showed that, （1） Glucose deprivation induced by MEM treat- ment for 6 h reduced survival rate and increased apoptotic rate in PC12 cells significantly compared to control group （P 〈 0.01）; and it produced a decrease in the expression of Glut-4 protein （P 〈 0.01）; （2） The expression level of ER-α36 was decreased significantly at 3 h of glucose deprivation, and then increased, while phosphorylation of Akt participated in the glucose deprivation was increased at first and then reduced; LY294002 （PI3K inhibitor） contributed to decreased expression of ER-α36, and suppressed the activation of Akt; （3） The rate of apoptosis was significantly increased in PC12-36L cells after glucose deprivation compared with that in wild type PC12 cells （P 〈 0.01）. Furthermore, phosphorylation of Akt was decreased and Caspase-3 was increased by glucose deprivation in PC12-36L cells com- pared with those in wild type PC12 cells. The study reveals that phosphorylation of Akt is associated with ER-α36 in PC12 cells exposed to glucose deprivation, and both are involved in the regulation of stress responses.