天然产物研究与开发 ›› 2016, Vol. 28 ›› Issue (11): 1687-1690.doi: 10.16333/j.1001-6880.2016.11.001

• 研究论文 •    下一篇

锁阳乙酸乙酯提取物的雌激素样作用研究

郑俊超1,马素亚1,于雪1,胡京红1,刘永刚2,田方泽2,鲁艺1*   

  1. 1北京中医药大学基础医学院;2北京中医药大学中药学院,北京 100029
  • 出版日期:2016-11-30 发布日期:2017-03-30

Estrogen-like Effect of Ethyl Acetate Extract of Cynomorium songaricum Rupr

ZHENG Jun-chao1,MA Su-ya1,YU Xue1,HU Jing-hong1,LIU Yong-gang2,TIAN Fang-ze2,LU Yi1*   

  1. 1 School of Preclinical Medicine;2 School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China
  • Online:2016-11-30 Published:2017-03-30

摘要: 雌激素提高学习记忆能力的功能已被广泛承认,前期实验证实锁阳乙酸乙酯提取物(ECS)可表现出雌激素样作用,提高去卵巢大鼠的学习记忆能力及血清E2水平,对Aβ-损伤SK-N-SH细胞有一定的保护作用。为了进一步研究ECS的雌激素样作用机制,拟在小鼠神经母细胞瘤细胞(Neuro2A)上探寻ECS对学习记忆相关蛋白突触素蛋白synaptophysin和环磷腺苷效应元件结合蛋白表达影响。采用CCK8法测定ECS对小鼠神经母细胞瘤细胞(Neuro2A)细胞活力的影响。将小鼠神经母细胞瘤细胞分为正常组、DMSO组、ECS组、Tamoxifen组、Tamoxifen+ECS组;正常组、ECS组、ECS+G15组和正常组、DMSO组、ECS组、PD98059组、 PD98059+ECS组。采用Western blot法测定synaptophysin,p-creb蛋白表达情况。实验表明锁阳乙酸乙酯提取物可促进synaptophysin,P-CREB表达,ER受体竞争性拮抗剂Tamoxifen可降低synaptophysin表达,对ECS促进synaptophysin表达无显著影响。GPR30受体特异性阻断剂G15可显著降低ECS促进synaptophysin表达。p-CREB上游Erk1/2-MAPK通路的阻断剂PD98059可显著抑制ECS上调p-CREB表达,结合前期实验结果,我们认为ECS可与GPR30结合通过MAPK通路促进学习记忆相关蛋白synaptophysin表达。

关键词: 锁阳, synaptophysin, p-CREB, GPR30

Abstract: It is widely recognized that estrogen can improve the ability of learning and memory function.Former study have shown that EtOAc extract of Cynomorium songaricum (ECS)can improve both E2 level and the ability of learning and memory in ovariectomized rat,protect skn-sh cells from Aβ- injury.To contribute a better understanding of the mechanism of ECS that improve ability of learning and memory,we investigated the expression of synaptophysin and p-CREB in Neuro-2a cells.Using cck8 to detect the influence of ECS on Neuro-2a cells.Divided the cells into Normal group,DMSO group,ECS group Tamoxifen group,ECS+Tamoxifen group;Normal group,ECS group,ECS+G15 group and Normal group,DMSO group,ECS group,PD98059 group,ECS+PD98059 group.The results showed that ECS can significantly improve the expression of synaptophysin and p-CREB on Neuro2A cells.Treatment of the cultures with ER antagonist Tamoxifen+ECS cannot significantly decreased the expression of synaptophysin,but with GPR30 antagonist G15+ECS can significantly decreased the expression of synaptophysin.The antagonist of MAPK/ERK pathway anyagonist PD98059+ECS also significantly decreased the expression of p-CREB.According to preliminary experimental results,we hypothesized that ECS can improve the ability of learning and memory through MAPK pathway by combing with GPR30.

Key words: Cynomorium songaricum, p-CREB, synaptophysin, GPR30

中图分类号: 

R285.5