蛋白激酶C活化在大鼠羟考酮耐受性形成中的作用及机制
Role and mechanism of protein kinase C activation in the development of oxycodone induced tolerance in rats
史艳华 1曾宾 1张治明 1徐巍 1肖裔兴 1邓厚盛1
作者信息
- 1. 郴州市第一人民医院麻醉科,郴州市 423000
- 折叠
摘要
目的 探讨蛋白激酶C活化在大鼠羟考酮耐受性形成中的作用及机制.方法 取雌性SD大鼠72只,体重250～280 g,按照随机数字表法将其随机分为Sham组、Sham+OXY组、Sham+PMA组、Sham+GÖ6983组、OXY+PMA组及OXY+GÖ6983组,每组12只.Sham组给予生理盐水,Sham+OXY组给予羟考酮,Sham+PMA组给予PKC激动剂(PMA),Sham+GÖ6983组给予PKC抑制剂(GÖ6983),OXY+PMA组给予羟考酮+PKC激动剂,OXY+GÖ6983组给予羟考酮+PKC抑制剂,均连续7 d.以von-Frey法评估每日最后一次给药后大鼠左侧后爪的机械缩足反应阈值(MWT).通过蛋白质印迹法和免疫荧光双标检测给药后第7日大鼠脊髓背角组织PKC及p-PKC的表达.结果 与Sham组相比,Sham+OXY组大鼠在每次给药30 min后,大鼠后爪MWT均上升(P均＜0.05);给药第7日脊髓背角组织PKC及p-PKC的表达均上升(P均＜0.05).免疫荧光双标显示p-PKC与脊髓背角组织的NeuN阳性细胞共定位表达.与Sham+OXY组相比,OXY+GÖ6983组大鼠在最后1次给药后4、8及16 h时MWT均上升(P均＜0.05),在第7日脊髓背角组织PKC及p-PKC的表达均下降(P均＜0.05);与Sham+OXY组相比,OXY+PMA组大鼠在最后一次给药后4、8、16h时间点MWT均下降(P均＜0.05),在第7日脊髓背角组织PKC及p-PKC的表达均上升(P均＜0.05).结论 大鼠腹腔长期注射OXY,可激活PKC引起耐受性形成,而联合注射OXY+GÖ6983可对其产生抑制作用.
Abstract
Objective To explore the role and mechanism of protein kinase C(PKC)activation in the development of oxycodone induced tolerance in rats.Methods Seventy-two female SD rats weighing 250-280 g,were randomly divided into Sham group,Sham+OXY group,Sham+PMA group,Sham+GÖ6983 group,OXY+PMA group and OXY+GÖ6983 group,12 rats in each group.Sham group given normal saline,Sham+OXY group given oxycodone(OXY),Sham+PMA group given PKC agonist(PMA),Sham+GÖ6983 group given PKC inhibitor(GÖ6983),OXY+PMA group given oxycodo ne+PKC agonist,OXY+GÖ6983 group given oxycodone+PKC inhibitor.The mechanical withdrawal threshold(MWT)of the left hind paw was evaluated by von-Frey method after the last daily administration.The expressions of PKC and p-PKC in spinal cord dorsal horn were detected by Western blot and immunofluorescence double-labeling on the 7th day after the administration.Results Compared with the Sham group,MWT increased at 30 minutes after each administration,the expressions of PKC and p-PKC in the dorsal horn of the spinal cord increased on the 7th day after the administration in the Sham+OXY group(all P＜0.05).Double immunofluorescence showed that p-PKC was co-localized with NeuN positive cells in the dorsal horn of the spinal cord.Compared with the Sham+OXY group,MWT increased at 4,8 and 16 h after the last administration,the expressions of PKC and p-PKC decreased on the 7th day after the administration in the OXY+GÖ6983 group(all P＜0.05).Compared with the Sham+OXY group,MWT decreased at 4,8 and 16 h after the last administration,the expressions of PKC and p-PKC in the spinal dorsal horn tissue increased on the 7th day after the administration in the OXY+PMA group(all P＜0.05).Conclusion Long-term intraperitoneal injection of OXY can induce tolerance in rats by activated PKC,and combined with injection of OXY+GÖ6983 can inhibit it.
关键词
羟考酮/蛋白激酶C/耐受性Key words
Oxycodone/Protein kinase C/Tolerance引用本文复制引用
基金项目
郴州市科技局科技发展计划(ZDYF2020031)
郴州市第一人民医院院级科研项目(N2020-6)
出版年
2024
NETL
NSTL
万方数据