CAJ | 학술논문

目的评价鞘内注射NR2B反义寡核苷酸对吗啡依赖大鼠认知功能的影响.方法健康雄性SD大鼠,体重230～270 g,腹腔注射60mg/kg戊巴比妥钠麻醉下,于L3,4间隙穿刺置管.取鞘内置管成功的大鼠30只,随机分为3组(n=10):对照组(C组)、吗啡依赖组(MD组)和NR2B反义寡核苷酸组(aNR2B组).MD组和aNR2B组皮下注射吗啡10 mg/kg,每天2次,隔天增加10mg/kg,至第6天时末次注射50mg/kg,建立吗啡依赖模型;C组皮下注射等容量生理盐水.造模完成后,MD组和aNR2B组每天上午8:00皮下注射吗啡30 mg/kg,连续注射4周;aNR2B组每天皮下注射吗啡前30 min时鞘内注射15 nmol NR2B反义寡核苷酸.于造模后给予吗啡即刻、1和3周时,采用Morris水迷宫进行认知功能测试,记录逃避潜伏期和跨越平台次数;然后处死大鼠,取海马,测定胆碱乙酰基转移酶(ChAT)的表达.结果与C组比较,造模后给予吗啡1、3周时MD组逃避潜伏期延长,跨越平台次数减少,海马ChAT表达下调,aNR2B组逃避潜伏期缩短,跨越平台次数增加,海马ChAT表达上调(P＜0.05);与MD组比较,造模后给予吗啡1、3周时aNR2B组逃避潜伏期缩短,跨越平台次数增加,海马ChAT表达上调(P＜0.05).结论鞘内注射NR2B反义寡核苷酸可减轻吗啡依赖大鼠认知功能障碍,其机制与上调海马ChAT的表达有关.
목적 평개초내주사NR2B반의과핵감산대마배의뢰대서인지공능적영향.방법 건강웅성SD대서,체중230～270 g,복강주사60mg/kg무파비타납마취하,우L3,4간극천자치관.취초내치관성공적대서30지,수궤분위3조(n=10):대조조(C조)、마배의뢰조(MD조)화NR2B반의과핵감산조(aNR2B조).MD조화aNR2B조피하주사마배10 mg/kg,매천2차,격천증가10mg/kg,지제6천시말차주사50mg/kg,건립마배의뢰모형;C조피하주사등용량생리염수.조모완성후,MD조화aNR2B조매천상오8:00피하주사마배30 mg/kg,련속주사4주;aNR2B조매천피하주사마배전30 min시초내주사15 nmol NR2B반의과핵감산.우조모후급여마배즉각、1화3주시,채용Morris수미궁진행인지공능측시,기록도피잠복기화과월평태차수;연후처사대서,취해마,측정담감을선기전이매(ChAT)적표체.결과 여C조비교,조모후급여마배1、3주시MD조도피잠복기연장,과월평태차수감소,해마ChAT표체하조,aNR2B조도피잠복기축단,과월평태차수증가,해마ChAT표체상조(P＜0.05);여MD조비교,조모후급여마배1、3주시aNR2B조도피잠복기축단,과월평태차수증가,해마ChAT표체상조(P＜0.05).결론 초내주사NR2B반의과핵감산가감경마배의뢰대서인지공능장애,기궤제여상조해마ChAT적표체유관.
Objective To evaluate the effect of intrathecal (IT) NR2B antisense oligonucleotide (aNR2B) on cognitive function in morphine-dependent rats.Methods Male SD rats weighing 230-270 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital 60 mg/kg.IT catheter was placed at L3-4 interspace according to the technique described by Yang. Thirty rats in which IT catheter was successfully placed without any complication were randomly divided into 3 groups(n=10 each):control group (group C), morphine dependence group (group MD) and group aNR2B.Morphine dependence was induced in group MD and aNR2B by increasing doses of morphine for 6 days. The initial dose of morphine was 10mg/kg injected subcutaneously (SC) twice a day and was increased by 10 mg/kg.every other day.The final dose was 50mg/kg. Then morphine 30 mg/kg was administered SC once a day for 4 weeks. aNR2B 15 nmol was administered IT at 30 min before SC morphine every day in group aNR2B.In control group normal saline was administered instead of morphine. Morris water maze was used to assess the cognitive function at 0 (T0, baseline),1 and 3 weeks of morphine administration (T1,T2).The escape latency and the number of times the animals crossing the plateform were recorded. The animals were killed after the test and the hippocampus was isolated for determination of choline acetytransferase(ChAT)expression.Results There was no significant difference in the baseline escape latency and the baseline number of times the animals crossing the plateform at T0 among the 3 groups. The escape latency was significantly prolonged and the number of times the animals crossing the plateform decreased at T1 and T2 as compared with the baseline at T0 in group MD.The ChAT expression was significantly down-regulated in group MD as compared with control group. IT aNR2B significantly ameliorated cognitive dysfunction at T1 and T2 and increased ChAT expression in group aNR2B compared with group MD.Conclusion IT NR2B antisense oligonucleotide can attenuate cognitive dysfunction through up-regulation of ChAT expression in hippocampus in morphine-dependent rats.