氢过氧化枯烯对SH-SY5Y细胞表达阿片受体介导的cAMP含量的影响  

Effect of cumene hydroperoxide on the cAMP formation in opioid receptors-responsive SH-SY5Y cells

作　　者：张玉柯[1] 王振华[1] 李德芳[1] 陈姬[1] 郑秋生[1] 

机构地区：[1]石河子大学药学院省部共建新疆特种植物药资源教育部重点实验室,石河子832002

出　　处：《中国新药杂志》2010年第16期1438-1442,共5页Chinese Journal of New Drugs

基　　金：国家自然科学基金(30860074);教育部新世纪优秀人才支持项目(NCET-06-0918)

摘　　要：目的:应用氢过氧化枯烯(cumene hydroperoxide,CHP)作用于体外经全反式维甲酸(all-transretinoic acid,RA)分化培养的人神经母细胞瘤细胞SH-SY5Y,观察其对阿片受体介导环磷酸腺苷(cyclic a-denosine monophosphate,cAMP)含量的影响。方法:CHP预处理分化SH-SY5Y细胞后,通过吗啡的急性、慢性作用和纳洛酮(naloxone,NLX)阻断,观察腺苷酸环化酶激动剂Forsklin(Fs)诱导的胞内cAMP水平的变化。不同浓度的CHP作用于分化的SH-SY5Y细胞24 h,进行细胞内活性氧(reactive oxygen species,ROS)水平、丙二醛(malondialdehyde,MDA)含量检测、超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活性检测。结果:吗啡急性作用,抑制了Fs诱导的胞内cAMP含量的增加,CHP预处理吗啡急性作用的SH-SY5Y细胞,显著降低了吗啡对Fs诱导的胞内cAMP水平的抑制作用;吗啡长期作用,使Fs诱导的胞内cAMP水平恢复到正常;NLX阻断,使胞内cAMP水平进一步升高。CHP预处理吗啡长期作用和NLX阻断的SH-SY5Y细胞,使吗啡对Fs诱导的胞内cAMP水平降低。CHP引起SH-SY5Y细胞的氧化损伤,使细胞内ROS的水平上升,MDA含量增高及降低细胞内SOD和CAT的活力。结论:ROS使μ-阿片受体的功能受到损伤,其可能通过调节阿片受体介导的cAMP含量的变化而参与了吗啡依赖的机制。Objective：To study the effect of cumene hydroperoxide（CHP） on the cAMP formation in opioid receptors-responsive all-trans retinoic acid（RA）-differentiated SH-SY5Y cells.Methods：SH-SY5Y cells were pretreated with CHP and were used to assess the effects of forskolin-induced changes of cAMP in human neuroblastoma SH-SY5Y cells exposed acutely or chronically to morphine（Mor） and withdrawal precipitated with naloxone（NLX）.The differentiated SH-SY5Y cells were treated by different concentrations of CHP for 24 h,the contents of reactive oxygen species（ROS） and malondiadehyde（MDA）,catalase（CAT） and superoxide dismutase（SOD） activation were measured.Results：Acute exposure of Mor resulted in Fs-induced inhibition of cAMP formation in SH-SY5Y cells,however,the exposure of cells to CHP prior to acute Mor treatment resulted in an increase in cAMP levels.Chronic Mor treatment,initially attenuated cAMP levels began to recover,but exposure of cells to CHP before chronic Mor treatment and NLX withdrawal resulted in an inhibition of cAMP formation.CHP was able to cause oxidative damage in SH-SY5Y cells,MDA contents and ROS levels were increased,CAT and SOD activation were reduced.Conclusion：Under conditions of oxidative damages,the function of μ-opioid receptor was significantly decreased.ROS changed the effects of opioid receptor agonists on intracellular cAMP,which associated with adaptive changes in Mor dependence.

关 键 词：吗啡 Μ-阿片受体 环磷酸腺苷 氢过氧化枯烯 

分 类 号：R964[医药卫生—药理学] R971[医药卫生—药学]