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作品数:6 被引量:4H指数:2
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功能化单壁碳纳米管/NR2B-shRNA干扰质粒复合物的制备及其用于慢性疼痛的基因治疗被引量:2
2013年
目的:评价鞘内注射功能化单壁碳纳米管shRNA干扰质粒复合物对福尔马林炎性痛小鼠的基因沉默效果。方法:先将整合了NR2B目的基因的pYr-1.1-hU6-EGFP(带绿色荧光)质粒进行扩增,抽提,消毒浓缩,浓度纯度测定,以获得高纯度、高浓度的NR2B-shRNA干扰质粒;将难溶于水的单壁碳纳米管用表面活性剂十六烷基三甲基溴化铵(CTAB)和超声振荡法制备成功能化单壁碳纳米管(f-SWNT)混悬液,通过凝胶成像系统检测功能化单壁碳纳米管与NR2B-shRNA的结合情况;选取雄性清洁级成年C57BL/6小鼠40只,体重18~23 kg,随机分为5组(n=8):空白组(C组),假手术组(S组),实验组(A组),阳性对照组(P组),阴性对照组(N组);分别行鞘内注射,7天后建立福尔马林炎性痛小鼠模型,测定术后1 h自发痛行为学改变,取脊髓L4~6节段采用RT-PCR法检测NR2BmRNA表达。结果:获得能够稳定表达的NR2B-shRNA干扰质粒;表面活性剂十六烷基三甲基溴化铵(CTAB)修饰单壁碳纳米管能形成较稳定的功能化单壁碳纳米管混悬液;功能化单壁碳纳米管能与NR2B-shRNA干扰质粒很好地结合,其结合最佳质量比为1:3.5;对于福尔马林所致的Ⅰ相急性痛,所有组间的急性期伤害性反应无差别(P〉0.05),阳性对照组(P组)和实验组(A组)小鼠对福尔马林所致的Ⅱ相持续痛的伤害性反应显著降低(P〈0.02);阳性对照组(P组)和实验组(A组)小鼠脊髓NR2BmRNA表达均低于其他对照组(P〈0.05)。结论:功能化单壁碳纳米管能与NR2B-shRNA干扰质粒很好地结合形成功能化单壁碳纳米管/NR2B-shRNA干扰质粒复合物;鞘内注射功能化单壁碳纳米管用作基因载体能有效转染NR2B-shRNA干扰质粒复合物行NR2B基因沉默,抑制小鼠脊髓NR2B基因表达,减轻福尔马林Ⅱ相痛行为学。
甘丽阎雪彬章饶香黄东
关键词:单壁碳纳米管炎性痛
Binding tendency with oligonucleotides and cell toxicity of cetyltrimethyl ammonium bromide-coated single-walled carbon nanotubes被引量:2
2011年
Functionalized carbon nanotubes (CNTs) were made for the delivery of genes and drugs and CNT-based biosensors. The basis of CNTs is for binding with biomolecules in biomedical applications. The binding tendency with small interfering RNA oligonucleotides and cytotoxicity of cetyltrimethyl ammonium bromide (CTAB)-coated single-walled carbon nanotubes (SWNTs) were studied. The field emission scanning electron microscopy and transmission electron microscopy results show that a SWNT suspension in CTAB solution was well-dispersed and stable. CTAB is the cross-linker between SWNTs and oligonucleotides. The CTAB-coated SWNTs have less cytotoxicity to human umbilical vein endothelial cells than single SWNTs and the cytotoxicity of CTAB-coated SWNTs depended on the concentration of CTAB-coated SWNTs.
阎雪彬谷永红黄东甘丽邬力翔黄利华陈哲东黄苏萍周科朝
关键词:OLIGONUCLEOTIDESCYTOTOXICITY
功能化单壁碳纳米管/NR2B-SiRNA复合物鞘内注射用于慢性疼痛基因治疗的研究
甘丽阎雪彬章饶香黄东
关键词:炎性痛
功能化单壁碳纳米管/NR2B-SiRNA复合物鞘内注射用于慢性疼痛基因治疗的研究
目的:制备功能化单壁碳纳米管混悬液,评价鞘内注射功能化单壁碳纳米管/NR2B-SiRNA复合物对福尔马林炎性痛小鼠的基因沉默效果。方法:先用表面活性剂十六烷基三甲基溴化铵(CTAB)和超声方法,分散修饰不溶于水的单壁碳纳...
甘丽阎雪彬章饶香黄东
关键词:炎性痛
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Cytotoxicity effect assessment of acid purified carbon nanotubes modified with cetyltrimethyl ammonium bromide
2013年
The cytotoxicities of single-walled carbon nanotubes (SWNTs) and acid purified single-walled carbon nanotubes (SWNT-COOH) were investigated by spectroscopic analysis. Cell viability and cell apoptosis were applied to assessing the cytotoxicity of SWNT-COOH, cetyltrimethyl ammonium bromide (CTAB) and acid purified carbon nanotubes modified with cetyltrimethyl ammonium bromide (SWNT-COOH/CTAB). The results indicate that SWNTs are more toxic than SWNT-COOH. Concentration and time-curve analyses indicate that cytotoxicity of SWNT-COOH/CTAB is more related to the toxicity of the surfactant CTAB. The cytotoxicity effect of CTAB and SWNT-COOH/CTAB is acceptable at low concentrations (0.5-25μg/mL). The cytotoxicity observation suggests that SWNT-COOH/CTAB can safely applied to biomedical field at low concentrations (0.5-25μg/mL).
甘丽阎雪彬杨金凤谷永红黄东章饶香黄利华
关键词:CYTOTOXICITYAPOPTOSIS
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