突触后致密物蛋白质95基因沉默及对神经病理性疼痛大鼠疼痛行为的干预

被引：12

作者：

李旭

申乐

许力

刘薇

虞雪融

黄宇光

机构：

[1] 中国医学科学院北京协和医学院北京协和医院麻醉科

来源：

协和医学杂志 | 2011年 / 2卷 / 04期

关键词：

神经病理性疼痛; 突触后致密物蛋白质95; 谷氨酸兴奋性中毒; Ca2+/钙调蛋白依赖的蛋白质激酶IIα; 慢性压迫损伤; 小干扰RNA;

D O I：

暂无

中图分类号：

R402 [];

学科分类号：

摘要：

目的评估小干扰RNA(small interfering RNA,siRNA)对突触后致密物蛋白质95(postsynaptic density protein95,PSD-95)基因的沉默效率及PSD-95基因沉默对谷氨酸诱导的神经细胞毒性与信号转导的影响,观察用RNA干扰(RNA interference,RNAi)技术沉默大鼠脊髓PSD-95基因治疗神经病理性疼痛的效果。方法用神经母细胞瘤/大鼠神经胶质细胞瘤杂交瘤细胞(NG108-15细胞)筛选大鼠PSD-95基因特异的siRNA,并用谷氨酸刺激PSD-95基因沉默的NG108-15细胞,检测细胞生长活力与信号通路蛋白质表达与磷酸化的改变。建立大鼠神经病理性疼痛的坐骨神经慢性压迫(chronic constrictioni njury,CCI)模型,预防性及治疗性鞘内给予PSD-95siRNA,留取脊髓标本,实时定量PCR(real-time PCR,RT-PCR)分析PSD-95 mRNA表达水平,并测定大鼠后足机械痛阈及热痛阈的变化。结果序列特异性最高的siRNA在最佳转染条件下使PSD-95基因表达水平下降91.5%;PSD-95基因沉默既可增强神经细胞对谷氨酸毒性的耐受能力,又可抑制Ca2+/钙调蛋白依赖的蛋白质激酶IIα(CaMKIIα)异构型磷酸化。正常大鼠鞘内注射PSD-95siRNA可降低大鼠脊髓背角PSD-95mRNA的表达水平38%(P<0.05),但对痛阈无显著影响;CCI模型大鼠鞘内注射PSD-95 siRNA可明显缓解神经病理性疼痛。结论 PSD-95基因在神经病理性疼痛的发生中起重要作用。鞘内注射PSD-95siRNA可以显著缓解CCI模型大鼠机械性和热痛觉过敏,PSD-95 siRNA可能成为有效控制神经病理性疼痛的基因治疗方法。

引用

页码：343 / 349

页数：7

共 14 条

[1]

Involvement of spinal phosphorylation cascade of Tyr1472-NR2B, Thr286-CaMKII, and Ser831-GluR1 in neuropathic pain [J].

Katano, Tayo ;

Nakazawa, Takanobu ;

Nakatsuka, Terumasa ;

Watanabe, Masahiko ;

Yamamoto, Tadashi ;

Ito, Seiji .

NEUROPHARMACOLOGY, 2011, 60 (04) :609-616

[2]

Impairment of CaMKII activation and attenuation of neuropathic pain in mice lacking NR2B phosphorylated at Tyr1472 [J].

Matsumura, Shinji ;

Kunori, Shunji ;

Mabuchi, Tamaki ;

Katano, Tayo ;

Nakazawa, Takanobu ;

Abe, Tetsuya ;

Watanabe, Masahiko ;

Yamamoto, Tadashi ;

Okuda-Ashitaka, Emiko ;

Ito, Seiji .

EUROPEAN JOURNAL OF NEUROSCIENCE, 2010, 32 (05) :798-810

[3]

Central Sensitization: A Generator of Pain Hypersensitivity by Central Neural Plasticity [J].

Latremoliere, Alban ;

Woolf, Clifford J. .

JOURNAL OF PAIN, 2009, 10 (09) :895-926

[4]

Intrathecal Raf-1-selective siRNA attenuates sustained morphine-mediated thermal hyperalgesia.[J].Suneeta Tumati;Tally Largent Milnes;Henry I. Yamamura;Todd W. Vanderah;William R. Roeske;Eva V. Varga.European Journal of Pharmacology.2008, 1

[5]

Small interfering RNA-mediated selective knockdown of Na V 1.8 tetrodotoxin-resistant sodium channel reverses mechanical allodynia in neuropathic rats.[J].X.-W. Dong;S. Goregoaker;H. Engler;X. Zhou;L. Mark;J. Crona;R. Terry;J. Hunter;T. Priestley.Neuroscience.2007, 2

[6]

Silencing of vanilloid receptor TRPV1 by RNAi reduces neuropathic and visceral pain in vivo [J].

Christoph, Thomas ;

Gruenweller, Arnold ;

Mika, Joanna ;

Schaefer, Martin K.-H. ;

Wade, Erik J. ;

Weihe, Eberhard ;

Erdmann, Volker A. ;

Frank, Robert ;

Gillen, Clemens ;

Kurreck, Jens .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2006, 350 (01) :238-243

[7]

Mechanisms of neuropathic pain [J].

Campbell, James N. ;

Meyer, Richard A. .

NEURON, 2006, 52 (01) :77-92

[8]

An Efficient Intrathecal Delivery of Small Interfering RNA to the Spinal Cord and Peripheral Neurons.[J].Miaw-Chyi Luo;Dong-Qin Zhang;Shou-Wu Ma;Yuan-Yuan Huang;Sam J Shuster;Frank Porreca;Josephine Lai.Molecular Pain.2005,

[9]

Suppression of postsynaptic density protein 95 by antisense oligonucleotides diminishes postischemic pyramidal cell death in rat hippocampal CA1 subfield.[J].Xiao-Yu Hou;Guang-Yi Zhang;De-Guang Wang;Qiu-Hua Guan;Jing-Zhi Yan.Neuroscience Letters.2005, 3

[10]

Neuropathic Sensitization of Behavioral reflexes and spinal NMDA receptor/CaM kinase II interactions are disrupted in PSD-95 mutant mice [J].

Garry, EM ;

Moss, A ;

Delaney, A ;

O'Neill, F ;

Blakemore, J ;

Bowen, J ;

Husi, H ;

Mitchell, R ;

Grant, SGN ;

Fleetwood-Walker, SM .

CURRENT BIOLOGY, 2003, 13 (04) :321-328

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