Inhibition of spinal protein kinase Cα expression by an antisense oligonucleotide attenuates morphine infusion-induced tolerance
Inhibition of spinal protein kinase Cα expression by an antisense oligonucleotide attenuates morphine infusion-induced tolerance
复制标题
DOI：
10.1016/s0306-4522(02)00157-4
复制
发表时间：
2002-01-01
期刊：
影响因子：
3.3
通讯作者：
Butler, M
中科院分区：
文献类型：
--
作者：
Hua, XY;Moore, A;Butler, M
包括α同工酶在内的蛋白激酶C同工型与吗啡耐受性有关。在本研究中，我们研究了鞘内递送针对大鼠蛋白激酶Calpha mRNA的反义寡核苷酸对脊髓蛋白激酶Calpha同工酶表达和脊髓吗啡耐受的影响。连续鞘内注入吗啡的大鼠产生的热刺激在第1天，这消失了第5天的缩爪阈值的增加。在第6天，推注鞘内注射吗啡（探针剂量）产生显着减少镇痛吗啡输注大鼠比盐水输注大鼠，表明耐受性。与脊髓吗啡输注同时进行的蛋白激酶Calpha反义的鞘内治疗不仅在5天输注期间维持了吗啡的镇痛作用，而且在第6天显著增加了对探针吗啡剂量的反应性。相比之下，在相同的治疗模式中使用的错误没有影响。蛋白激酶Calpha反义核酸对脊髓吗啡耐受的抑制作用呈剂量依赖性，且可逆。实时定量逆转录-聚合酶链反应和蛋白质印迹显示，鞘内治疗的反义，但不是错义，在大鼠脊髓蛋白激酶Calpha mRNA和蛋白质的表达降低。γ同工酶的表达不受寡核苷酸的影响。反义还减弱脊髓中蛋白激酶C介导的磷酸化。这些结果表明，脊髓蛋白激酶Calpha同工酶的表达的选择性减少，然后减少局部蛋白激酶C介导的磷酸化将逆转脊髓吗啡输注诱导的耐受性。这一发现与吗啡输注产生的耐受依赖于蛋白激酶C磷酸化的增加的观点一致，也强调了蛋白激酶Calpha同工酶及其在脊髓中的激活可能特异性地参与阿片耐受现象。(C)2002年IBRO。由爱思唯尔科技有限公司出版。保留所有权利。
Protein kinase C isoforms including the alpha isozyme have been implicated in morphine tolerance. In the present study, we examined the effect of intrathecal delivery of an antisense oligonucleotide targeting rat protein kinase Calpha mRNA on the expression of spinal protein kinase Calpha isozyme and spinal morphine tolerance. Continuous intrathecal infusion of rats with morphine produced an increase in paw withdrawal threshold to thermal stimulation on day 1, which disappeared by day 5. On day 6, a bolus intrathecal injection of morphine (a probe dose) produced significantly less analgesia in morphine-infused rats than in saline-infused rats, suggesting tolerance. Intrathecal treatment with the protein kinase Calpha antisense concurrent with spinal morphine infusion not only maintained the analgesic effect of morphine during the 5-day infusion, it also significantly increased responsiveness to the probe morphine dose on day 6. In comparison, the missense used in the same treatment paradigm had no effect. The inhibitory effect of protein kinase Calpha antisense on spinal morphine tolerance was dose-dependent, and reversible. Intrathecal treatment with the antisense, but not the missense, in rats decreased expression of spinal protein kinase Calpha mRNA and protein, as revealed by real-time quantitative reverse transcription-polymerase chain reaction and western blots. Expression of the gamma isozyme was not affected by the oligonucleotides. The antisense also attenuated protein kinase C-mediated phosphorylation in spinal cord. These results demonstrate that selective reduction in the expression of the spinal protein kinase Calpha isozyme followed by a decrease of local protein kinase C-mediated phosphorylation will reverse spinal morphine infusion-induced tolerance. This finding is consistent with the view that tolerance produced by morphine infusion is dependent upon an increase in phosphorylation by protein kinase C, and also it emphasizes that the protein kinase Calpha isozyme and its activation in spinal cord may specifically participate in the phenomenon of opiate tolerance. (C) 2002 IBRO. Published by Elsevier Science Ltd. All rights reserved.