中国组织工程研究

中国组织工程研究 ›› 2013, Vol. 17 ›› Issue (12): 2185-2190.doi: 10.3969/j.issn.2095-4344.2013.12.014

• 药物控释材料 drug delivery materials • 上一篇    下一篇

水溶性脂质体运载基因药物复合物治疗大鼠神经病理性痛

胡春奎1，陆建华1，陈 昊1，熊家祥2   

  1. 1解放军广州军区广州总医院麻醉科，广东省广州市 510010
    2解放军第三军医大学生理教研室，重庆市 400038
  • 收稿日期:2012-06-01 修回日期:2012-07-02 出版日期:2013-03-19 发布日期:2013-03-19
  • 通讯作者: 陆建华，博士后，解放军广州军区广州总医院麻醉科，广东省广州市 510010 ljianh@yahoo.com.cn
  • 作者简介:胡春奎★，女，1985年生，江西省婺源县人，汉族，南方医科大学在读硕士，主要从事慢性疼痛的基因治疗研究。 huchunkui2@163.com

Water-soluble lipopolymer carrying gene drugs for treatment of neuropathic pain in rats

Hu Chun-kui1, Lu Jian-hua1, Chen Hao1, Xiong Jia-xiang2   

  1. 1 Department of Anesthesiology, Guangzhou General Hospital of Guangzhou Military Command of Chinese PLA, Guangzhou 510010, Guangdong Province, China
    2 Department of Physiology, Third Military Medical University, Chongqing 400038, China
  • Received:2012-06-01 Revised:2012-07-02 Online:2013-03-19 Published:2013-03-19
  • Contact: Lu Jian-hua, Doctor, Department of Anesthesiology, Guangzhou General Hospital of Guangzhou Military Command of Chinese PLA, Guangzhou 510010, Guangdong Province, China ljianh@yahoo.com.cn
  • About author:Hu Chun-kui★, Studying for master’s degree, Department of Anesthesiology, Guangzhou General Hospital of Guangzhou Military Command of Chinese PLA, Guangzhou 510010, Guangdong Province, China huchunkui2@163.com

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摘要：

背景：有研究报道病毒载体运载N-甲基-D天冬氨酸受体1小干扰RNA可有效缓解大鼠炎性疼痛，但病毒载体存在安全隐患。
目的：探讨水溶性脂质体运载N-甲基-D天冬氨酸受体1小干扰RNA在体内外沉默N-甲基-D天冬氨酸受体1的效应和治疗神经病理性痛的可行性。
方法：将PC12随机分为阴性转染组、对照转染组和水溶性脂质体转染组，分别以N-甲基-D-天冬氨酸受体1 小干扰RNA、聚乙烯亚胺与N-甲基-D-天冬氨酸受体1 小干扰RNA的复合物及水溶性脂质体与N-甲基-D-天冬氨酸受体1 小干扰RNA的复合物转染PC12细胞，检测各组N-甲基-D-天冬氨酸受体1 基因mRNA及蛋白水平表达的变化。将48只SD大鼠随机分为假手术组、模型组、聚乙烯亚胺组及水溶性脂质体组，后3组建立大鼠神经病理性疼痛模型，并分别鞘内注射生理盐水、聚乙烯亚胺与N-甲基-D-天冬氨酸受体1 小干扰RNA的复合物和水溶性脂质体与N-甲基-D-天冬氨酸受体1 小干扰RNA的复合物；假手术组只暴露坐骨神经。
结果与结论：水溶性脂质体转染组N-甲基-D-天冬氨酸受体1的 mRNA与蛋白表达水平明显低于其他两组(P < 0.01)。与假手术组比较，模型组、聚乙烯亚胺组及水溶性脂质体组N-甲基-D-天冬氨酸受体1 的mRNA和蛋白表达上调，累积疼痛评分升高(P < 0.01)；与模型组比较，水溶性脂质体转染组脊髓背角N-甲基-D-天冬氨酸受体1 mRNA与蛋白表达及累积疼痛评分下降(P < 0.01)，聚乙烯亚胺组上述指标无明显变化(P > 0.05)。表明在体内条件下水溶性脂质体可有效运载N-甲基-D-天冬氨酸受体1 小干扰RNA，抑制N-甲基-D-天冬氨酸受体1 的过度表达，还可减轻大鼠神经病理性痛。

关键词: 生物材料, 生物材料与药物控释, 水溶性脂质体, 载体药物, N-甲基-D天冬氨酸受体1, 小干扰RNA, PC12细胞, 神经病理性痛, 省级基金\生物材料图片文章

Abstract:

BACKGROUND: Some studies have shown that viral vectors can carry N-methyl-D-aspartic acid receptors 1 (NR1) small interference RNA (siRNA) to relieve inflammatory pain in rats, but the viral vectors are unsafe.
OBJECTIVE: To examine the potential application of a non-viral gene carrier, water soluble lipopolymer (WSLP) for delivering siRNA targeting NR1 in vitro and in vivo and to determine whether WSLP-NR1siRNA complexes can be a new method for neuropathic pain treatment.
METHODS: PC12 cells were randomly divided into three groups: group WSLP-negative NR1 siRNA (negative group), group polyethylenimine-NR1 siRNA (control transfection group) and group WSLP- NR1 siRNA (group WS). NR1 expressions were detected using reverse transcription-PCR and western blot analysis. Forty-eight healthy male Sprague-Dawley rats were randomly divided into four groups (n=12 in each group): sham operation group (sham group), neuropathic pain group (model group), group polyethylenimine-NR1 siRNA (group PEI) and group WSLP- NR1 siRNA (group WSLP). Neuropathic pain models were established in the latter three groups. Normal saline, PEI-NR1 siRNA complex and WSLP-NR1 siRNA were injected intrathecally in model, PEI and WSLP groups, respectively, at 1 day after operation. Only the sciatic nerve was exposed in the sham group.
RESULTS AND CONCLUSION: NR1 mRNA and protein expression significantly decreased in group WS as compared with negative and control transfection groups (P < 0.01). Compared with the sham group, NR1 mRNA and protein expression significantly increased in the model, PEI and WSLP groups, and cumulative pain scores in the latter three groups were also higher (P < 0.01). Compared with the model group, intrathecal injection of WSLP- NR1siRNA complexes could relieve neuropathic pain and inhibit NR1 gene expression with reductions in mRNA and protein levels (P < 0.01), and intrathecal injection of PEI-NR1siRNA complexes did not show this inhibitory effect (P > 0.05). These results demonstrate that WSLP not only efficiently delivers NR1 siRNA targeting NR1 in vivo and inhibits the expression of NR1, but also reduces neuropathic pain in rats.

Key words: biomaterials, biomaterials and controlled drug release, water-soluble liposomes, carrier drugs, N-methyl-D-aspartate receptor 1, small interfering RNA, PC12 cells, neuropathic pain, provincial grants-supported paper, biomaterial photographs-containing paper

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