蛋白激酶Cα相互作用蛋白（PICK1）-AMPA受体运输在瑞芬太尼痛觉过敏中的研究

Remifentanil induced hyperalgesia has become one of the most important challeges anesthetists face for it's converse results. Spinal central sensitization via NMDA receptors and protein kinase C is involved in the mechanism of remifenanil indeced hyperalgsia.Our previors studies have shown that the increased trafficking of NMDA receptor NR2B subunit in the post-synaptic membrane of spinal cord neurons is involved in the mechanism of remifenanil indeced hyperalgsia. .Our preliminary experiments have shown that the increased activity of AMPA receptor is involved in the mechanism of remifenanil indeced hyperalgsia. PICK1 is an important protein which connects PKCa with AMPA receptor in the post-synaptic membrane.We suppose that PICK1 is involved in the mechanism of remifenanil indeced hyperalgsia and act as a bridge between NMDA receptor and AMPA receptor via NMDA receptor-PKCa- PICK1-AMPA receptor pathway.From cellular to animal levels, using technologies such as behavioral tests, Western Blot, Immunofluorescence , whole cell patch-clamp,PICK1 genetic knockout and PICK1 antisense oligodeoxynucleotide knockdown approach, we characterized the functional role of PICK1 in remifentanil induced hyperalgesia. Our study will provide a new target for the treatment and prevention of remifentanil-induced hyperalgesia.

瑞芬太尼诱发的痛觉过敏由于与其治疗初衷相悖，成为麻醉医生面临的巨大挑战之一。其发生机制与NMDA受体-蛋白激酶C（PKC）通路介导的脊髓中枢敏化相关。本课题组前期研究发现，脊髓突触后膜NMDA受体NR2B亚单位跨膜运输增加参与瑞芬太尼痛觉过敏的发生。预实验发现，同为离子型谷氨酸受体的AMPA受体也参与了瑞芬太尼痛觉过敏的发生。蛋白激酶C相互作用蛋白（PICK1）是PKCa和突触后膜AMPA受体间的衔接蛋白，因此我们认为PICK1可能参与瑞芬太尼痛觉过敏的发生，并且通过NMDA受体-PKCa- PICK1-AMPA受体通路成为连接NMDA受体和AMPA受体的桥梁。本项目拟从离体和动物水平，利用膜片钳技术、行为学实验、Western Blot、免疫荧光、PICK1基因敲除及反义核苷酸抑制等方法，探讨PICK1在瑞芬太尼痛觉过敏中的作用和机制，为预防和治疗瑞芬太尼痛觉过敏提供新的靶点。

瑞芬太尼诱发的痛觉过敏由于与其治疗初衷相悖，成为麻醉医生面临的巨大挑战之一。其发生机制与NMDA受体-蛋白激酶C（PKC）通路介导的脊髓中枢敏化相关。预实验发现，同为离子型谷氨酸受体的AMPA受体也参与了瑞芬太尼痛觉过敏的发生。 蛋白激酶C相互作用蛋白（PICK1）是PKCa和突触后膜AMPA受体间的衔接蛋白，本课题就PICK1–AMPA受体运输通路在瑞芬太尼痛觉过敏发生机制中的作用进行了深入的研究。首先，建立大鼠瑞芬太尼痛觉过敏模型，应用PICK1基因反义寡核苷酸抑制、免疫荧光和western blot 技术，通过观察PICK1基因反义寡核苷酸抑制对APMA受体运输的影响，在体证明了PICK1–AMPA受体运输通路在瑞芬太尼痛觉过敏发生机制中的关键作用；其次，建立腰段脊髓切片瑞芬太尼痛觉过敏模型，应用PICK1拮抗剂、免疫荧光和全细胞膜片钳技术，通过观察PICK1拮抗剂对APMA受体电生理功能和运输的影响，离体证明了PICK1–AMPA受体运输通路在瑞芬太尼痛觉过敏发生机制中的关键作用。上述结果为预防和治疗瑞芬太尼痛觉过敏提供新的靶点。