脊髓PKCε和μ阿片受体参与瑞芬太尼诱导大鼠痛觉过敏  

Roles of Spinal PKC ond MOR in Remifentanil Induced Hyperalgesia in Rats

作　　者：于萍 王晓娥 崔宇[3] 陈元[3] YU Ping;WANG Xiao-e;CUI Yu;CHEN Yuan(Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 510080,China;Department of Anesthesiology,The First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080,China;School of Medicine,Sun Yat-sen University,Guangzhou 510080,China)

机构地区：[1]中山大学中山医学院,广东广州510080 [2]中山大学附属第一医院麻醉科,广东广州510080 [3]中山大学医学院,广东广州510080

出　　处：《中山大学学报（医学科学版）》2020年第6期850-857,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省重点领域研发计划项目(2018B030335001)。

摘　　要：【目的】在瑞芬太尼诱导痛觉过敏的大鼠脊髓组织中观察PKCε和μ阿片受体(MOR)的表达水平变化,探讨PKCε和MOR两者之间的关系。【方法】18只成年雄性SD大鼠随机平均分为3组(每组6只),分为空白对照组(C组)、生理盐水对照组(S组)和瑞芬太尼组(R组)。通过大鼠尾静脉输注瑞芬太尼4μg/(kg·min)给药2 h建立瑞芬太尼诱导的大鼠痛觉过敏模型。采用Hargreaves法观察瑞芬太尼输注后6、24 h的大鼠热刺激撤足阈值。通过荧光定量PCR和蛋白免疫印迹法检测大鼠脊髓组织中的PKCε和MOR的表达水平,并用免疫荧光观察PKCε和MOR在大鼠脊髓组织中的分布情况。瑞芬太尼输注前30 min预先鞘内注射不同的PKC抑制剂Bisindolylmaleimide I(BIS,PKC广谱抑制剂)和Gö6983(抑制PKC的亚型不包括PKCε),观察不同的PKC抑制剂预处理对瑞芬太尼诱导的大鼠痛觉过敏的影响以及检测大鼠脊髓组织MOR蛋白表达变化。【结果】输注瑞芬太尼24 h后可显著引起大鼠热痛阈值降低[PWTL=(5.31±0.87)s,P<0.01]。同时,蛋白免疫印迹和实时荧光定量PCR结果表明,PKCε表达上调(P<0.001)、MOR表达下降(P<0.001)。免疫荧光结果表明,在大鼠脊髓背角PKCε和MOR均与神经元存在共定位。预先鞘内注射BIS可以缓解大鼠痛觉过敏以及抑制MOR表达上调(P<0.001),而注射Gö6983则无影响(P>0.05)。【结论】脊髓背角神经元PKCε和MOR参与瑞芬太尼诱导大鼠痛觉过敏,且PKCε可能调控MOR表达水平变化,本研究将为探明瑞芬太尼诱导痛觉过敏机制提供新的理论依据。【Objective】To observe the expression of PKCεandμopioid receptor(MOR)and explore their relationship in the spinal cord of rats with remifentanil-induced hyperalgesia(RIH).【Methods】Eighteenadult male SD rats were randomly divided into control group,saline group and remifentanil group with 6 rats in each group.Hyperalgesia was induced by continuous infusion of 4μg/(kg·min)remifentanil for 2 hours via vein tail.Pain sensitivity was assessed by the Hargreaves test to determine paw withdrawl latency to a thermal stimulus at 6 h and 24 h after remifentanil administration.Real-time fluorescent quantitative PCR and Western blot analysis were used to measure the expression levels of PKCεand MOR in the spinal cord of rats and their distribution was detected by immunofluorescence.In order to observe the effect of inhibitors on RIH and the protein expression of MOR in the spinal cord of rats,we injected different PKC inhibitors including Bisindolylmaleimide I(BIS,inhibiting PKCε)and Gö6983(not inhibiting PKCε)intrathecally 30 mins prior to remifentanil infusion.【Results】The results of behavioral experiment showed that the paw withdrawal thermal latency was significantly reduced at 24 h after remifentanil infusion[PWTL=(5.31±0.87)s,P<0.01].Western blot and Real time PCR results revealed upregulated PKCεexpression(P<0.001)and downregulated MOR expression(P<0.001).Immunofluorescence results indicated that PKCεand MOR were colocalized with neurons in the dorsal horn of rat spinal cord.Preemptive intrathecal injection of BIS alleviated the hyperalgesia in rats and inhibited MOR expression(P<0.001),but injection of Gö6983made no difference(P>0.05).【Conclusion】PKCεand MORin spinal dorsal horn neurons may be involved in RIH,and PKCεmay regulate the expression levels of MOR.This research provides a new theoretical mecha⁃nism for the prevention and treatment of RIH.

关 键 词：瑞芬太尼 痛觉过敏 PKCΕ Μ阿片受体 

分 类 号：R338.3[医药卫生—人体生理学]