NR2B-siRNA Mediated by Hydroxyapatite Nanoparticles Relieves for Malin-Induced Pain of Mice

Abstract:

To observe the effect of NR2B-siRNA mediated by hydroxyapatite nanoparticles (HA) on formalin-induced inflammatory pain of mice and the expression of NR2B in spinal cord. To preliminarily investigate the feasibility of HA as siRNA carrier to transfer NR2B-siRNA in vivo. The sequence-specific NR2B-siRNA of mice was designed and synthesized initially. Using HA as a siRNA carrier, green fluorescent protein(GFP)-siRNA as the control, 4 ug of NR2B-siRNA was administered into subarachnoid space of mice via conscious injection. On 7th day after intrathecal injection, formalin test was observed for 1 hour in each group, followed by dissection of lumbar segments of spinal cords immediately for use in immunohistochemical staining of NR2B. The results show that NR2B-siRNA not only significantly abolish the nociceptive response of mice in the tonic phase induced by formalin, but also decrease the amount of cells expressing NR2B protein in spinal cord, while GFP-siRNA mediated by HA don’t produce the same effects, which demonstrates that HA is capable of effectively transfering NR2B-siRNA via intrathecal injection, furthermore, HA/NR2B-siRNA complex can significantly reduce formalin-induced pain of mice, and specificly inhibit NR2B expression in spinal cord of mice.

• Periodical: Advanced Materials Research (Volumes 343-344)
• Pages: 926-932
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.343-344.926
• Online since: September 2011
• Authors: Yong Hong Gu, Xue Bin Yan, Dong Huang, Rui Han, Li Xiang Wu
• Keywords: Formalin Pain, Hydroxyapatite (HAP), Mice, Nanoparticle, NR2B, Small Interfering RNA

References

[1] Ma Qp, Hargreaves RJ. Localization of N-methyl-d-aspartate NR2B subunits on primary sensory neurons that give rise to small-caliber sciatic nerve fibers in rats. Neuroscience, 2000, 101(3): 699~707.

DOI: 10.1016/s0306-4522(00)00419-x

[2] Annika B. Malmberg, Heather Gilbert, R. Tyler McCabe, et al. Powerful antinociceptive effects of the cone snail venom-derived subtype-selective NMDA receptor antagonists conantok ins G and T . Pain, 2003, 101(1-2): 109~116.

DOI: 10.1016/s0304-3959(02)00303-2

[3] Nagy GG, Watanabe M, Fukaya M, etal. Synaptic distribution of the NR1, NR2A and NR2B subunits of the N-methyl-d-aspartate receptor in the rat lumbar spinal cord revealed with an antigen-unmasking technique. Eur J Neurosci 2004, 20(12): 3301~3312.

DOI: 10.1111/j.1460-9568.2004.03798.x

[4] Garraway SM, Xu Q, Inturrisi CE. Design and evaluation of small interfering RNAs that target expression of the N-methyl-D-aspartate receptor NR1 subunit gene in the spinal cord dorsal horn. Journal of Pharmacology and experimental Therapeutics , 2007, 322(3): 982~988.

DOI: 10.1124/jpet.107.123125

[5] Tan PH, Yang LC, Shin HC, etal Gene knockdown with intrathecal siRNA of NMDA receptor NR2B subunit reduces formalin-induced nociception in the rat. Gene Therapy, 2005, 12(1): 59~66.

DOI: 10.1038/sj.gt.3302376

[6] Gabriele Dorn, Sadhana Patel, Glen Wotherspoon, et al. SiRNA relieves chronic neuropathic pain. Nucleic Acids Research, 2004, 32(5): e49.

DOI: 10.1093/nar/gnh044

[7] X. -W. Dong, S. Goregoaker, H. Engler, et al. Small interfering RNA-mediated selective knockdown of NaV1. 8 tetrodotoxin-resistant sodium channel reverses mechanical allodynia in neuropathic rats. Neuroscience, 2007, 146(2): 812~821.

DOI: 10.1016/j.neuroscience.2007.01.054

[8] Zhu shaihong, Huang Boyun, Zhou Kechao, Huang Suping, Liu Fang, Li Yimin, Xue Zigang, Long Zigao . Hydroxyapatite Nanoparticles as a Novel Gene Carrier. Journal of Nanoparticle Research 2004; 6: 311~317.

DOI: 10.1023/b:nano.0000034721.06473.23

[9] JIANG Ming, ZHANG Yong-quan, HE Guang-xiang, et al. Protective effect of NT-3 gene mediated by hydroxyapatite nanoparticle on the cochlea of guinea pigs injured by excitotoxicity. J Cent South Univ (Med Sci), 2007, 32(4): 563-567 (Language: Chinese).

[10] Elbashir SM, Harborth J, Weber K, Tuschl T. Analysis of gene function in somatic ammalian cells using small interfering RNAs. Methods 2002, 26: 99~213.

DOI: 10.1016/s1046-2023(02)00023-3

[11] Zimmerman M. Ethicalguide lines for investigations of experimental pain in conscious animals. Pain, 1983; 16: 109~110.

[12] Fairbanks FC. Spinal delivery of analgesics in experimental models of Pain and analgesia. Advanced Drug Delivery Reviews, 2003, 55(8): 1007~1041.

DOI: 10.1016/s0169-409x(03)00101-7

[13] IshizakiK, SasakiM, etal. The effect to fintrathecal magnesium sulphate on nociception in rat acute Pain models. Anaesthesia, 1999, 54: 241~246.

DOI: 10.1046/j.1365-2044.1999.00741.x

[14] Brummelkamp TR, Bernards R, Agami R. Stable suppression of Tumorigenicity by virus-mediated RNA interference. Cancer Cell, 2002, 243~247.

DOI: 10.1016/s1535-6108(02)00122-8

[15] Devroe E, Silver PA. Retrovirus-delivered siRNA. BMC Biotechnol, 2002, 2: 15.