PKMζ—Kalirin-7—AMPA受体GluR1膜上位在瑞芬太尼二次痛敏中的调控机制

瑞芬太尼痛敏后可形成痛敏记忆，二次手术再次使用瑞芬太尼会引发更严重的二次痛敏，其机制尚不明确。课题组已发表工作证实NMDA和AMPA受体均参与瑞芬太尼痛敏形成和维持, 其中AMPA受体GluR1膜上位与瑞芬太尼痛敏密切相关。新近文献报道PKMζ调控AMPA受体活化可维持脊髓记忆功能LTP产生，课题组也发现PKMζ可调节瑞芬太尼二次痛敏大鼠Kalirin-7表达，故推测PKMζ是介导瑞芬太尼二次痛敏的关键。本研究拟建立大鼠在体瑞芬太尼二次痛敏模型，利用痛敏行为学测定、Western blot、共沉淀、荧光和高尔基染色研究PKMζ、Kalirin-7、GluR1和树突棘形态在瑞芬太尼二次痛敏中动态变化；体外培养大鼠脊髓片和背角神经元，抑制PKMζ和Kalirin-7研究PKMζ—Kalirin-7—GluR1膜上位在瑞芬太尼二次痛敏中的机制。本研究将为瑞芬太尼二次痛敏的临床防治提供新思路。

Remifentanil-induced hyperalgesia is followed by hyperalgesic memory, a second surgery with remifentanil anesthesia is identified to generate greater secondary postoperative hyperalgesia, but its mechanism remains elusive. Our previous studies have manifested that NMDA and AMPA receptors are involved in development and maintenance of remifentanil induced hyperalgesia, moreover, GluR1 membrane insertion is associated with remifentanil-induced hyperalgesia. Recent studies have reported that protein kinase M zeta (PKMζ) mediated AMPA receptor activation is critical for formation of spinal cord memory function— LTP. We also have found that PKMζ could modulate Kalirin-7 expression in remifentanil-induced secondary hyperalgesia in rats. We hypothesize that PKMζ may exert a pivotal effect on remifentanil-induced secondary hyperalgesia. Therefore in this study, following accomplishment of a rat model of remifentanil-induced secondary hyperalgesia, we use nociceptive behavior test, Western blot, co-immunoprecipitation, immunofluorescence and Golgi staining to detect dynamic changes of PKMζ, Kalirin-7, GluR1 and dendritic spines structure in remifentanil-induced secondary hyperalgesia. We also culture rat primary spinal dorsal horn neurons and spinal cord slice, and inhibit PKMζ and Kalirin-7 to demonstrate the role of PKMζ— Kalirin-7—GluR1 membrane insertion in remifentanil-induced secondary hyperalgesia. The present study will provide the novel options for the effective prevention of secondary hyperalgesia induced by remifentanil in clinics.