国际麻醉学与复苏杂志
國際痳醉學與複囌雜誌
국제마취학여복소잡지
INTERNATIONAL JOURNAL OF ANESTHESIOLOGY AND RESUSCITATION
2013年
4期
316-319,338
,共5页
杨雪%董文芳%彭捷%熊家祥%陆建华
楊雪%董文芳%彭捷%熊傢祥%陸建華
양설%동문방%팽첩%웅가상%륙건화
水溶性脂聚体%N-甲基-D-天冬氨酸受体受体2B%小干涉RNA%炎性疼痛
水溶性脂聚體%N-甲基-D-天鼕氨痠受體受體2B%小榦涉RNA%炎性疼痛
수용성지취체%N-갑기-D-천동안산수체수체2B%소간섭RNA%염성동통
Water soluble lipopolymer%N-Methyl-D-aspartic acid receptor 2B%Small interfering RNA%Chronic inflammatory pain
目的 慢性炎性疼痛的基因治疗缺乏安全且高效的载体,本实验探讨了低分子量聚乙烯亚胺(polyethylenimine,PEI)和胆固醇组成的水溶性脂聚体(water-soluble lipopolymer,WSLP)运载N-甲基-D-天冬氨酸受体2B亚基(N-Methyl-D-aspartic acid receptor 2B,NR2B)小干涉RNA(small interfering RNA,siRNA)治疗大鼠慢性炎性疼痛的可行性. 方法 将WSLP直接与靶向NR2B的siRNA连接形成WSLP/siRNA复合物,乱序siRNA作为对照(WSLP/scRNA)；然后检测鞘内注射WSLP/siRNA对完全弗氏佐剂(freund's adjuvant complete,CFA)炎性痛模型大鼠脊髓背角(spinal cord doral horn,SCDH)NR2B蛋白表达的影响,并且测定鞘内注射此复合物后炎性痛大鼠机械缩足反射阈值(mechanical withdrawal threshold,MWT)及热缩足反射持续时间(thermal withdrawal duration,TWD)的变化. 结果 WSLP/siRNA注射后3d,炎性疼痛大鼠SCDH NR2B蛋白水平的表达与CFA组比较显著下降58％(P＜0.01),WSLP/scRNA及PEI/siRNA注射后NR2B的表达无明显变化(P＞0.05).WSLP/siRNA鞘内注射后3、4d及5d炎性痛大鼠的MWT [(3 d:(9.2±1.3)；4 d:(9.8±1.2)；5 d:(9.5±1.2)]较CFA组[(3 d:(4.6±1.2)；4 d:(4.9±1.8)；5 d:(5.0±1.8)]显著增高(P＜0.01),TWD[(3 d:(3.27±0.32)s；4 d:(3.83±0.49)s；5 d:(3.57±0.33)s]较CFA组[(3 d:(6.71±0.45)s；4 d:(6.97±0.54)s；5 d:(6.63±0.38)s]显著缩短(P＜0.01),WSLP/scRNA及PEI/siRNA均无此治疗作用(P＞0.05). 结论 WSLP可有效运载siRNA抑制CFA所致慢性炎性痛大鼠NR2B的过度表达,从而对炎性疼痛大鼠具有治疗作用.
目的 慢性炎性疼痛的基因治療缺乏安全且高效的載體,本實驗探討瞭低分子量聚乙烯亞胺(polyethylenimine,PEI)和膽固醇組成的水溶性脂聚體(water-soluble lipopolymer,WSLP)運載N-甲基-D-天鼕氨痠受體2B亞基(N-Methyl-D-aspartic acid receptor 2B,NR2B)小榦涉RNA(small interfering RNA,siRNA)治療大鼠慢性炎性疼痛的可行性. 方法 將WSLP直接與靶嚮NR2B的siRNA連接形成WSLP/siRNA複閤物,亂序siRNA作為對照(WSLP/scRNA)；然後檢測鞘內註射WSLP/siRNA對完全弗氏佐劑(freund's adjuvant complete,CFA)炎性痛模型大鼠脊髓揹角(spinal cord doral horn,SCDH)NR2B蛋白錶達的影響,併且測定鞘內註射此複閤物後炎性痛大鼠機械縮足反射閾值(mechanical withdrawal threshold,MWT)及熱縮足反射持續時間(thermal withdrawal duration,TWD)的變化. 結果 WSLP/siRNA註射後3d,炎性疼痛大鼠SCDH NR2B蛋白水平的錶達與CFA組比較顯著下降58％(P＜0.01),WSLP/scRNA及PEI/siRNA註射後NR2B的錶達無明顯變化(P＞0.05).WSLP/siRNA鞘內註射後3、4d及5d炎性痛大鼠的MWT [(3 d:(9.2±1.3)；4 d:(9.8±1.2)；5 d:(9.5±1.2)]較CFA組[(3 d:(4.6±1.2)；4 d:(4.9±1.8)；5 d:(5.0±1.8)]顯著增高(P＜0.01),TWD[(3 d:(3.27±0.32)s；4 d:(3.83±0.49)s；5 d:(3.57±0.33)s]較CFA組[(3 d:(6.71±0.45)s；4 d:(6.97±0.54)s；5 d:(6.63±0.38)s]顯著縮短(P＜0.01),WSLP/scRNA及PEI/siRNA均無此治療作用(P＞0.05). 結論 WSLP可有效運載siRNA抑製CFA所緻慢性炎性痛大鼠NR2B的過度錶達,從而對炎性疼痛大鼠具有治療作用.
목적 만성염성동통적기인치료결핍안전차고효적재체,본실험탐토료저분자량취을희아알(polyethylenimine,PEI)화담고순조성적수용성지취체(water-soluble lipopolymer,WSLP)운재N-갑기-D-천동안산수체2B아기(N-Methyl-D-aspartic acid receptor 2B,NR2B)소간섭RNA(small interfering RNA,siRNA)치료대서만성염성동통적가행성. 방법 장WSLP직접여파향NR2B적siRNA련접형성WSLP/siRNA복합물,란서siRNA작위대조(WSLP/scRNA)；연후검측초내주사WSLP/siRNA대완전불씨좌제(freund's adjuvant complete,CFA)염성통모형대서척수배각(spinal cord doral horn,SCDH)NR2B단백표체적영향,병차측정초내주사차복합물후염성통대서궤계축족반사역치(mechanical withdrawal threshold,MWT)급열축족반사지속시간(thermal withdrawal duration,TWD)적변화. 결과 WSLP/siRNA주사후3d,염성동통대서SCDH NR2B단백수평적표체여CFA조비교현저하강58％(P＜0.01),WSLP/scRNA급PEI/siRNA주사후NR2B적표체무명현변화(P＞0.05).WSLP/siRNA초내주사후3、4d급5d염성통대서적MWT [(3 d:(9.2±1.3)；4 d:(9.8±1.2)；5 d:(9.5±1.2)]교CFA조[(3 d:(4.6±1.2)；4 d:(4.9±1.8)；5 d:(5.0±1.8)]현저증고(P＜0.01),TWD[(3 d:(3.27±0.32)s；4 d:(3.83±0.49)s；5 d:(3.57±0.33)s]교CFA조[(3 d:(6.71±0.45)s；4 d:(6.97±0.54)s；5 d:(6.63±0.38)s]현저축단(P＜0.01),WSLP/scRNA급PEI/siRNA균무차치료작용(P＞0.05). 결론 WSLP가유효운재siRNA억제CFA소치만성염성통대서NR2B적과도표체,종이대염성동통대서구유치료작용.
Objective To examine the potential application of a non-viral gene carrier,water soluble lipopolymer (WSLP)for delivering siRNA targeting NMDA receptor 2B (NR2B) in vivo and to determin whether WSLP/siRNA complexes can be a new method for chronic inflammatory pain treatment.Methods WSLP was complexed with siRNA (designed to inhibit NR2B expression)or scrambled siRNA (as a control).Changes of NR2B expression were detected using western-blot in SCDH of chronic inflammatory pain rats following intrathecal injection of WSLP/siRNA.Pain control efficacy was evaluated by changes of mechanical withdrawal threshold (MWT) and thermal withdrawal duration (TWD) in these rats.Results NR2B protein expression was efficiently inhibited by intrathecal injection of WSLP/siRNA complexes,and the changes of protein level was reduce by 58％ compared to chronic inflammatory pain rats without treatment (P＜0.01),while complexes injection of WSLP with scrambled siRNA or PEI with siRNA did not show this inhibitory effect (P＞0.05).Moreover,injection of WSLP/siRNA complexes significantly reduced MWT [(3 d:(9.2±1.3),4 d:(9.8±1.2),5 d:(9.5±1.2)] and increased TWD [(3 d:(3.27±0.32) s,4 d:(3.83±0.49) s,5 d:(3.57±0.33) s] of chronic inflammatory pain rats accordingly compared to chronic inflammatory pain rats without treatment [(MWT:3 d:(4.6±1.2); 4 d:(4.9±1.8),5 d:(5.0±1.8); TWD:3 d:(6.71±0.45) s,4 d:(6.97±0.54) s,5 d:(6.63±0.38) s] (P＜0.01).Conclusions These results demonstrate that WSLP can efficiently deliver siRNA targeting NR2B in vivo and relieve chronic inflammatory pain.
