CAJ | 학술논문

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%生物材料图片文章生物材料%生物材料與藥物控釋%水溶性脂質體%載體藥物%N-甲基-D 天鼕氨痠受體 1%小榦擾 RNA%PC12 細胞%神經病理性痛%省級基金%生物材料圖片文章
생물재료%생물재료여약물공석%수용성지질체%재체약물%N-갑기-D 천동안산수체 1%소간우 RNA%PC12 세포%신경병이성통%성급기금%생물재료도편문장
biomaterials%biomaterials and control ed drug release%water-soluble liposomes%carrier drugs%N-methyl-D-aspartate receptor 1%smal interfering RNA%PC12 cells%neuropathic pain%provincial grants-supported paper%biomaterial photographs-containing paper

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.
배경:유연구보도병독재체운재 N-갑기-D 천동안산수체1소간우 RNA 가유효완해대서염성동통,단병독재체존재안전은환.목적:탐토수용성지질체운재 N-갑기-D 천동안산수체1소간우 RNA 재체내외침묵 N-갑기-D 천동안산수체1적효응화치료신경병이성통적가행성.방법:장 PC12수궤분위음성전염조、대조전염조화수용성지질체전염조,분별이 N-갑기-D-천동안산수체1소간우 RNA、취을희아알여 N-갑기-D-천동안산수체1소간우 RNA 적복합물급수용성지질체여 N-갑기-D-천동안산수체1소간우 RNA 적복합물전염 PC12세포,검측각조 N-갑기-D-천동안산수체1기인 mRNA 급단백수평표체적변화.장48지 SD 대서수궤분위가수술조、모형조、취을희아알조급수용성지질체조,후3조건립대서신경병이성동통모형,병분별초내주사생리염수、취을희아알여N-갑기-D-천동안산수체1소간우 RNA 적복합물화수용성지질체여 N-갑기-D-천동안산수체1소간우 RNA 적복합물;가수술조지폭로좌골신경.결과여결론:수용성지질체전염조 N-갑기-D-천동안산수체1적 mRNA 여단백표체수평명현저우기타량조(P <0.01).여가수술조비교,모형조、취을희아알조급수용성지질체조 N-갑기-D-천동안산수체1적 mRNA 화단백표체상조,루적동통평분승고(P <0.01);여모형조비교,수용성지질체전염조척수배각 N-갑기-D-천동안산수체1 mRNA 여단백표체급루적동통평분하강(P <0.01),취을희아알조상술지표무명현변화(P >0.05).표명재체내조건하수용성지질체가유효운재 N-갑기-D-천동안산수체1소간우 RNA,억제 N-갑기-D-천동안산수체1적과도표체,환가감경대서신경병이성통.
BACKGROUND: Some studies have shown that viral vectors can carry N-methyl-D-aspartic acid receptors 1 (NR1) smal 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 intrathecal y 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.