疼痛基础研究（中青年优秀论文评奖）.docVIP

疼痛基础研究 （中青年优秀论文评奖） 侧脑室注射β-arrestin2抗基因RNA慢病毒对小鼠吗啡镇痛和耐受的影响 刘希江 卜慧莲 曹菲 杨辉 高峰( 田玉科 华中科技大学同济医学院附属同济医院麻醉学教研室，武汉，430030 摘要 目的 探讨侧脑室注射慢病毒介导的抗基因RNA下调小鼠脑内β-arrestin2表达的可行性及其对吗啡镇痛作用和耐受形成的影响。方法 将与β-arrestin2基因转录起始位点互补的抗基因RNA序列构建入慢病毒载体pGC-FU，将该质粒与病毒包装质粒一同转染293T细胞，包装成表达β-arrestin2抗基因RNA的重组慢病毒。将该慢病毒注射到小鼠侧脑室，用免疫组化和免疫印迹方法鉴定该重组慢病毒的基因沉默效率，并观察其对小鼠吗啡镇痛作用和吗啡耐受形成的影响。结果 免疫组化和免疫蛋白印迹证实小鼠侧脑室注射抗基因RNA 慢病毒可显著抑制小鼠脑内β-arrestin2蛋白的表达，蛋白抑制率为7%。与对照组相比，侧脑室注射β-arrestin2抗基因RNA慢病毒后，小鼠吗啡镇痛作用显著增强（P0.05）且吗啡镇痛剂量依赖性显著增强（P0.05）。同时侧脑室注射β-arrestin2抗基因RNA慢病毒可有效阻止小鼠吗啡慢性耐受的形成（P0.05）。结论 侧脑室注射β-arrestin2抗基因RNA慢病毒可显著抑制小鼠脑内β-arrestin2蛋白的表达，增强吗啡镇痛作用，有效阻止小鼠吗啡慢性耐受的形成。 关键词：吗啡；镇痛；耐受；beta-arrestin2；基因沉默 Effects of intracerebroventricular microinjection of β-arrestin2 antigene RNA lentiviruses on mice morphine analgesia and tolerance LIU Xi-jiang, BU Hui-lian, CAO Fei, YANG Hui, GAO Feng and TIAN Yu-ke Department of Anesthesiology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China Corresponding author: GAO Feng, Department of Anesthesiology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China. E-mail: gaofeng9261@ Abstract Objective To investigate the feasibility of inhibiting β-arrestin2 expression through intracerebroventricular microinjection of β-arrestin2 antigene RNA lentiviruses and observe its impact on morphine analgesia and tolerance in mice. Method The RNA sequence complementary to β-arrestin2 gene transcription start site was constructed into the lentiviral vector pGC-FU. Then, this plasmid and viral packaging plasmids were co-transfected into 293T cells to produce recombinant lentiviruses that expressing β-arrestin2 antigene RNAs. The β-arrestin2 antigene RNA expressing lentiviruses were stereotacticly microinjected into lateral cerebral ventricles of mice. Immunohistochemistry and western blot were adopted to identify the gene silencing efficiency. Additionally, effects of int