水通道4基因敲除对吗啡诱导小鼠条件性位置偏爱及神经发生的影响  

Effect of aquaporin 4 gene knockout on morphine-induced conditioned place preference and neurogenesis

作　　者：王志媛 吴宁 李锦 WANG Zhi-yuan;WU Ning;LI Jin(Beijing Key Laboratory of Neuropsychopharmacology,State Key Laboratory of Toxicology and Medical Countermeasures,Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences,Beijing 100850,China)

机构地区：[1]军事科学院军事医学研究院毒物药物研究所,抗毒药物与毒理学国家重点实验室,神经精神药理学北京市重点实验室,北京100850

出　　处：《中国药理学与毒理学杂志》2021年第8期575-581,共7页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的研究水通道4(AQP4)基因敲除对吗啡(Mor)诱导的小鼠条件性位置偏爱(CPP)及神经发生的影响。方法取野生型(WT)和AQP4基因敲除型(KO)小鼠,分别连续8 d sc给予生理盐水(WT-Con和KO-Con组)或Mor 10 mg·kg^(-1)(WT-Mor和KO-Mor组),同时训练小鼠,建立小鼠CPP模型。CPP表达测试后,ip给予5′-溴脱氧尿嘧啶(BrdU),24 h后采用免疫组化法检测小鼠海马齿状回的颗粒细胞层下区(SGZ)神经前体细胞增殖(BrdU+);28 d后,采用BrdU/神经元特异性核抗原(NeuN)免疫荧光双染法检测神经前体细胞的存活(BrdU+)和向神经元的分化(BrdU+/NeuN+)。结果在CPP模型上,小鼠伴药箱停留时间WT-Mor组和KO-Mor组分别显著高于WT-Con组和KO-Con组(P<0.05,P<0.01),KO-Mor组显著低于WT-Mor组(P<0.05),WT-Con组与KO-Con组无显著差异。免疫组织化学染色实验结果显示,小鼠海马SGZ区BrdU+细胞数WT-Mor组较WT-Con组减少了(28.3±21.3)%(P<0.05),而KO-Mor组与KO-Con组间、WT-Con组与KO-Con组间和WT-Mor与KO-Mor组间均无显著性差异。免疫荧光双染结果显示,给予BrdU 28 d后,小鼠海马SGZ区BrdU+细胞数WT-Mor组显著低于WT-Con组(P<0.05),KO-Mor组与WT-Mor组间、KO-Con组与WT-Con组间和KO-Mor组与KO-Con组间均无显著差异;而BrdU+细胞存活率KO-Mor组〔(60.3±10.1)%〕较WT-Mor组〔(35.3±9.3)%〕显著升高(P<0.05),WT-Con组与KO-Con组间、WT-Mor组与WT-Con组间和KO-Mor组与KO-Con组间均无显著差异;BrdU+NeuN+细胞数WT-Mor组较WT-Con组显著降低(P<0.05),KO-Mor组较WT-Mor组显著升高(P<0.05),WT-Con组与KO-Con组间和KO-Mor组与KO-Con组间均无显著差异;神经元分化率各组间均无显著性差异。结论AQP4基因敲除可降低Mor诱导小鼠CPP效应的形成,并抵抗Mor慢性处理对海马神经发生的抑制,其机制与减轻Mor对神经前体细胞增殖、存活和向神经元分化的抑制有关。OBJECTIVE To investigate the effects of aquaporin 4(AQP4)gene knockout on conditioned place preference(CPP)and neurogenesis induced by morphine(Mor).METHODS Wild-type(WT)and AQP4 knockout(KO)mice were sc injected with saline(WT-Con and KO-Con groups)or Mor 10 mg·kg^(-1)(WT-Mor and KO-Mor groups)for 8 consecutive days to establish a CPP model.After CPP test,5′-bromodeoxyuracil(BrdU)was ip administered.After 24 h,BrdU immunohistochemical staining was used to study the effect of AQP4 gene knockout on the negative regulation of neural progenitor cell proliferation(BrdU+)in the subgranular zone(SGZ)of dentate gyrus in the hippocampus by Mor.After 28 d,BrdU immunofluorescence staining was used to detect the effect of AQP4 gene knockout on the negative regulation of survival(BrdU+)and differentiation(BrdU+NeuN+)of neural progenitor cells in SGZ by Mor.RESULTS The WT-Mor group and the KO-Mor group spent a much longer time in the drug-associated compartment than the WT-Con group and the KO-Con group(P<0.01,P<0.05)respectively,but the KO-Mor group spent much less time in the drug-associated compartment than the WT-Mor group(P<0.05).The time spent in the drug-associated compartment was not significantly different between the WT-Con group and the KO-Con group.The results of immunohistochemical staining experiments showed that the number of BrdU+cells in SGZ in the WT-Mor group was reduced by as much as(28.3±21.3)%compared with the WT-Con group(P<0.05).But there was no significant difference between the WT-Con group and the KO-Con group or between the KO-Mor group and the KO-Con group.The results of immunofluorescence staining experiments showed that after 28 days of administration of BrdU,the number of BrdU+cells in SGZ in the WT-Mor group was significantly smaller than that of the WT-Con group(P<0.05).There was no significant difference between WT-Con and KO-Con groups or between the KO-Mor and KO-Con groups or between KO-Mor and WT-Mor groups.The survival rate of the KO-Mor group(60.3±10.1)%was significantly higher th

关 键 词：水通道蛋白4 吗啡 条件性位置偏爱 神经发生 成瘾记忆 

分 类 号：R964[医药卫生—药理学]