CAJ | 학술논문

目的探讨水溶性脂聚体(WSLP)介导含2B亚基的N-甲基-D-天冬氨酸受体小干扰RNA(NR2B siRNA)治疗大鼠神经病理性痛的可行性.方法健康雄性SD大鼠100只,周龄6周,体重180～200 g,采用随机数字表法,将大鼠随机分为5组(n=20):对照组(C组)、假手术组(S组)、神经病理性痛组(NP组)、WSLP-NR2B siRNA组(W组)、WSLP-阴性对照NR2B siRNA(WN组).采用坐骨神经分支部分结扎法制备大鼠神经病理性痛模型.C组不予任何处理;S组仅暴露坐骨神经,不牵拉和损伤神经;NP组于制备模型后即刻鞘内注射生理盐水20μl;W组和WN组于制备模型后即刻分别鞘内注射相应的siRNA 20μl.于模型制备前1 d及制备后3、7、14和21 d时测定机械缩足反应阈值(MWT)及热缩足反应持续时间(TWD),模型制备后3 d,痛阈测定结束后,每组取10只大鼠,取L4-6节段背根神经节,测定NR2B mRNA及其蛋白的表达水平.结果与S组比较,NP组、W组和WN组MWT降低,TWD延长,NR2B mRNA及其蛋白表达上调(P＜0.05或0.01),C组上述指标差异无统计学意义(P＞0.05);与NP组比较,W组MWT升高,TWD缩短,NR2B mRNA及其蛋白表达下调(P＜0.01),WN组上述指标差异无统计学意义(P＞0.05).结论 WSLP不仅成功介导NR2B siRNA,抑制NR2B的表达,还可减轻大鼠神经病理性痛.
목적 탐토수용성지취체(WSLP)개도함2B아기적N-갑기-D-천동안산수체소간우RNA(NR2B siRNA)치료대서신경병이성통적가행성.방법 건강웅성SD대서100지,주령6주,체중180～200 g,채용수궤수자표법,장대서수궤분위5조(n=20):대조조(C조)、가수술조(S조)、신경병이성통조(NP조)、WSLP-NR2B siRNA조(W조)、WSLP-음성대조NR2B siRNA(WN조).채용좌골신경분지부분결찰법제비대서신경병이성통모형.C조불여임하처리;S조부폭로좌골신경,불견랍화손상신경;NP조우제비모형후즉각초내주사생리염수20μl;W조화WN조우제비모형후즉각분별초내주사상응적siRNA 20μl.우모형제비전1 d급제비후3、7、14화21 d시측정궤계축족반응역치(MWT)급열축족반응지속시간(TWD),모형제비후3 d,통역측정결속후,매조취10지대서,취L4-6절단배근신경절,측정NR2B mRNA급기단백적표체수평.결과 여S조비교,NP조、W조화WN조MWT강저,TWD연장,NR2B mRNA급기단백표체상조(P＜0.05혹0.01),C조상술지표차이무통계학의의(P＞0.05);여NP조비교,W조MWT승고,TWD축단,NR2B mRNA급기단백표체하조(P＜0.01),WN조상술지표차이무통계학의의(P＞0.05).결론 WSLP불부성공개도NR2B siRNA,억제NR2B적표체,환가감경대서신경병이성통.
Objective To investigate the feasibility of NR2B small interference RNA(NR2B siRNA)carried by water-soluble lipopolymer(WSLP)for treatment of neuropathic pain in rats.Methods One hundred healthy male SD rats weighing 180-200 g were randomly divided into 5 groups(n=20 each):normal control group (group C),sham operation group(group S),neuropathic pain group(group NP),group WSLP-NR2B siRNA (group W)and group WSLP-negative NR2B siRNA(group WN).Neuropathic pain was induced by partial ligation of sciatic nenre.WSLP-NR2B siRNA complex was formed by binding WSLP and NR2B siRNA.Normal saline.WSLP-NR2B siRNA complex and WSLP-negative NR2B siRNA 20μl were injected intrathecally after operation in NP,W and WN groups respectively.Mechanical withdrawal threshold(MWT)and thermal withdrawal duration (TWD)were measured before(baseline)and at 3,7,14 and 21 days after operation.Ten animals in each group were sacrificed on the 3rd day after operation and the lumbar segment(L4-6)of the dorsal root ganglia was removed for determination of the expression of NR2B mRNA and protein using RT-PCR and Western blot analysis.Results Sciatic nerve ligation significantly decreased MWT and prolonged TWD and increased NR2B mRNA and protein expression in group NP as compared with group C.WSLP-NR2B siRNA complex significantly reduced sciatic nerve ligation-induced hyperalgesia and decreased NR2B mRNA and protein expression in group W as compared with group NP.Conclusion WSLP not only mediates NR2B siRNA successfully and inhibits the expression of NR2B,but also reduces neuropathic pain in rats.