Down-regulation of Stargazin inhibits the enhanced surface delivery of α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor GluR1 subunit in rat dorsal horn and ameliorates postoperative pain.
Down-regulation of Stargazin inhibits the enhanced surface delivery of α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor GluR1 subunit in rat dorsal horn and ameliorates postoperative pain.
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DOI：
10.1097/aln.0000000000000291
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发表时间：
2014-09
期刊：
影响因子：
8.8
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Guo R;Zhao Y;Zhang M;Wang Y;Shi R;Liu Y;Xu J;Wu A;Yue Y;Wu J;Guan Y;Wang Y
Stargazin是已知的第一个调节α-氨基-3-羟基-5-甲基-4-异恶唑丙酸酯(AMPA)受体突触靶向的跨膜蛋白。然而，目前尚不清楚Stargazin对脊髓AMPA受体亚单位表面递送的调节是否有助于术后疼痛的发生。Western印迹分析检测AMPA受体亚基GluR1和GluR2在大鼠背角表面递送的变化。用免疫共沉淀法检测了Stargazin与GluR1和GluR2的相互作用。鞘内注射小干扰核糖核酸(SiRNA)311可抑制Stargazin的表达。切开后3h(β微管蛋白的1.49±0.15倍)和1d(1.03±0.25)，与对照组(0.63±0.23，P<；0.05，n=6)相比，同侧背角膜结合型GluR_1含量增加(1.49±0.15倍，P=0.05)。切开后3h，GluR1与Stargazin免疫共沉淀量(1.48±0.31倍)明显高于对照组(0.45±0.24，P<0.05，n=6)。重要的是，鞘内注射Stargazin siRNA311(0.87±0.09)可使膜GluR_1在切开后3h升高至接近对照组的水平(0.72±0.20倍)，但不包括干扰siRNA(1.48±0.24)或赋形剂(1.25±0.13，P<；0.05，n=6/组)。Stargazin siRNA311预处理可阻止Stargazin-GluR1相互作用的增加，并减轻术后疼痛。这项研究表明，Stargazin介导的GluR1亚单位表面递送在术后疼痛的发生中起着关键作用。对于术后疼痛，更好的治疗策略可能包括选择性地下调脊髓星状氮素，以抑制突触靶向GluR1亚单位。
Stargazin is the first transmembrane protein known to regulate synaptic targeting of α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors. Yet, it is unclear whether regulation of the surface delivery of spinal AMPA receptor subunits by stargazin contributes to postoperative pain development. Western blot analysis was used to examine changes in the surface delivery of AMPA receptor subunits GluR1 and GluR2 in rat dorsal horn. The interaction between stargazin and GluR1 and GluR2 was examined by coimmunoprecipitation. Expression of stargazin was suppressed by intrathecal administration of small interfering ribonucleic acid (siRNA)311. Membrane-bound GluR1, but not GluR2, in ipsilateral dorsal horn was increased at 3 h (1.49±0.15-fold of β-tubulin, mean±SEM) and 1 day (1.03±0.25) after incision, as compared to that in control rats (naïve, 0.63±0.23, P < 0.05, n=6/group). The amount of GluR1 coimmunoprecipitated with stargazin was greater at 3 h after incision (1.48±0.31-fold of input) than in control animals (0.45±0.24, P < 0.05, n=6/group). Importantly, the increase in membrane GluR1 at 3 h after incision was normalized to near control level (0.72±0.20-fold of β-tubulin) by pretreatment with intrathecal stargazin siRNA311 (0.87±0.09), but not scrambled siRNA (1.48±0.24) or vehicle (1.25±0.13, P < 0.05, n=6/group). Stargazin siRNA311 pretreatment prevented the increase in stargazin–GluR1 interaction and decreased postoperative pain after incision. This study suggests a critical role of stargazin-mediated surface delivery of GluR1 subunit in the development of postoperative pain. A better therapeutic strategy for postoperative pain may involve selectively downregulating spinal stargazin to inhibit synaptic targeting of GluR1 subunit.