活性氧在阿片代谢过程中产生机制及其对阿片受体结合能力的影响

项目名称： 活性氧在阿片代谢过程中产生机制及其对阿片受体结合能力的影响

项目编号： No.30860074

项目类型： 地区科学基金项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 郑秋生

作者单位： 石河子大学

项目金额： 25万元

中文摘要： 本研究以神经细胞为研究对象，通过建立人神经母细胞瘤SH-SY5Y细胞吗啡依赖模型模型，采用激光共聚焦显微镜方法，研究吗啡依赖过程中活性氧产生机制，明确吗啡诱导细胞产生活性氧的种类和机制；弄清活性氧对阿片受体结合能力的影响，揭示活性氧与吗啡依赖和戒断等过程的作用。通过选用新疆特色的植物药有效成分干预吗啡依赖的研究，为新疆特种植物药和维药的研究积累资料。

中文关键词： 海洛因；自由基；活性氧；抗氧化；阿片受体

英文摘要： Object: To study the roles of ROS in intracellular ROS levels and in the expression of Mu opioid receptor during morphine chronic addiction., to demonstrate the special relationship between ROS and morphine addiction. Methods： (1) All-trans-retinoic acid (RA) was used to treat the human neuroblastoma (SH-SY5Y) cells for 6 days in order to get the differentiated cells with the property of neurons. (2) Spectrophotometric method was used to evaluated lactate dehydrogenase (LDH) viability and NBT reductive ability. (3) When the differentiated SH-SY5Y cells were treated with morphine in different concentration gradients and in different time periods, the level of ROS in the SH-SY5Y cells was measured by fluorescent probe, the expression of μpioid receptor (hMOPr) mRNA in the SH-SY5Y cells was reflected by RT-PCR. (4) As SH-SY5Y cells were pretreated with CHP, enzyme linked immunosorbent assay (ELISA) was used to assess the cAMP levels in human neuroblastoma SH-SY5Y cells exposured acutely or chronically to morphine and withdrawal precipitated with naloxone hydrochloride After exogenous oxidants and antioxidants were used to treat the differentiated SH-SY5Y cells, the level of ROS in the SH-SY5Y cells was measured by fluorescent probe, the expression of μpioid receptor (hMOPr) mRNA in the SH-SY5Y cells was reflected by RT-PCR. Results： (1) The SH-SY5Y cells treated by 10 μRA displayed the the property of neurons.. (2) There was a certain positive correlation between the generation of ROS and morphine addiction and the level of ROS could influence the expression of human μpioid receptor (hMOPr). (3) Exposured to acute morphine resulted in an inhibition of cAMP formation induced by Fs in SH-SY5Y cells while exposured to CHP prior to acute morphine treatment resulted in an increase of cAMP levels. Treated by chronic morphine, the initial attenuated cAMP levels began to recover, but exposured to CHP before chronic morphine treatment and acute Nal withdrawal resulted in an inhibition of cAMP formation. (3) The addition of exogenous oxidants was able to increase the generation of ROS in further in SH-SY5Y cells and antioxidants were opposite to that. Conclusion： (1)There was a certain positive correlation between the generation of ROS and morphine addiction. relationship between morphine chronic dependent and the generation of ROS and the (2) The exogenous oxidants and antioxidants were able to influence the effect of morphine addiction by affecting the expression of μpioid receptor (hMOPr) mRNA in SH-SY5Y cells. (3) The exogenous oxidants was able to influence the effect of morphine addiction by affecting the cAMP formation. in SH-SY5Y cells..(4) Antioxidant agents may hold the potentiality to reverse the effect of morphine addiction in the human neuroblastoma SH-SY5Y cells.

英文关键词： morphine addiction；Mu opioid receptor；ROS；oxidant/antioxidant；hMOPr mRNA