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
2011年
6期
683-686,694
,共5页
吗啡依赖%NR2B反义寡核苷酸%μ受体%κ受体
嗎啡依賴%NR2B反義寡覈苷痠%μ受體%κ受體
마배의뢰%NR2B반의과핵감산%μ수체%κ수체
Morphine dependence%NR2B antisense oligonucleotide%μ opiate receptor%κ opiate receptor
目的 观察NR2B反义寡核苷酸(NR2B antisense oligonucleotide,ANR2B)对吗啡依赖大鼠脊髓和脑干μ受体与κ受体表达的影响,探讨N-甲基-D-天冬氨酸(N-methyl-D-aspartic acid,NMDA)受体的NR2B亚基与阿片μ受体与κ受体在介导吗啡依赖和耐受过程中的相互作用.方法 雄性SD大鼠,体重230 g～270 g.鞘内埋管成功大鼠40只,按随机数字表法分为5组,每组8只.吗啡皮下注射首日10 mg/kg,每日2次,隔日每次增加10 mg/kg,至第6天末次注射50 mg/kg,建立吗啡依赖模型,以纳洛酮4 mg/kg单次腹腔注射建立吗啡依赖催促戒断模型,观察纳洛酮催促戒断症状并评价造模是否成功.分为吗啡依赖组(A组)；相应时间点皮下注射等量生理盐水为空白对照组(B组)；末次注射吗啡后4h腹腔注射盐酸纳洛酮4 mg/kg激发戒断症状为吗啡戒断组(C组)；在皮下注射吗啡的同时鞘内注射15 nmol ANR2B为吗啡+ANR2B组(D组)；在纳洛酮激发前30 min鞘内注射15 nmol ANR2B为戒断+ANR2B组(E组).应用RT-PCR检测吗啡依赖与戒断大鼠脊髓和脑干μ受体与κ受体mRNA表达.结果 ①吗啡依赖大鼠脊髓和脑干NR2B mRNA表达(0.965±0.012；0.958±0.020)较对照组(0.763±0.031)明显上调,NR2B反义寡核苷酸使其表达明显下调(0.786±0.011).②吗啡戒断症状主要表现为湿狗样摇体、激惹、齿颤、眼睑下垂、腹泻等.鞘内注射NR2B反义寡核苷酸则明显减轻戒断症状的总评分(32±2,22±3).③吗啡依赖大鼠脊髓和脑干中μ受体mRNA表达(0.955±0.021,0.946±0.013)较对照组升高有统计学意义.鞘内注射ANR2B则明显降低吗啡依赖大鼠脊髓和脑干中μ受体mRNA的表达(0.801±0.013;0.824±0.021).纳洛酮激发戒断症状后1h,脊髓和脑干中μ受体mRNA的表达较吗啡依赖组明显下降.激发前30 min鞘内注射ANR2B,脊髓和脑干中μ受体mRNA表达与戒断组比较变化无统计学意义.④吗啡依赖大鼠脊髓和脑干中κ受体mRNA表达(0.735±0.025；0.757±0.010)较对照组(0.974±0.016；0.982±0.013)降低有统计学意义；同时鞘内注射ANR2B则显著增加脊髓和脑干中κ受体mRNA的表达(0.875±0.012; 0.854±0.016)；纳洛酮激发戒断症状后1h,脊髓和脑干中κ受体mRNA的表达较吗啡依赖组明显增高；激发前30min鞘内注射ANR2B,脊髓和脑干中κ受体mRNA表达与戒断组比较变化无统计学意义.各组中β -actin表达量差异无统计学意义.结论 脊髓和脑干中μ受体与κ受体表达的变化在吗啡依赖与戒断反应中发挥了重要作用.鞘内注射NR2B反义寡核苷酸通过有效逆转吗啡依赖与戒断大鼠脊髓和脑干中μ受体与κ受体表达的变化趋势,从而发挥其抑制吗啡依赖与减轻戒断症状的作用.
目的 觀察NR2B反義寡覈苷痠(NR2B antisense oligonucleotide,ANR2B)對嗎啡依賴大鼠脊髓和腦榦μ受體與κ受體錶達的影響,探討N-甲基-D-天鼕氨痠(N-methyl-D-aspartic acid,NMDA)受體的NR2B亞基與阿片μ受體與κ受體在介導嗎啡依賴和耐受過程中的相互作用.方法 雄性SD大鼠,體重230 g～270 g.鞘內埋管成功大鼠40隻,按隨機數字錶法分為5組,每組8隻.嗎啡皮下註射首日10 mg/kg,每日2次,隔日每次增加10 mg/kg,至第6天末次註射50 mg/kg,建立嗎啡依賴模型,以納洛酮4 mg/kg單次腹腔註射建立嗎啡依賴催促戒斷模型,觀察納洛酮催促戒斷癥狀併評價造模是否成功.分為嗎啡依賴組(A組)；相應時間點皮下註射等量生理鹽水為空白對照組(B組)；末次註射嗎啡後4h腹腔註射鹽痠納洛酮4 mg/kg激髮戒斷癥狀為嗎啡戒斷組(C組)；在皮下註射嗎啡的同時鞘內註射15 nmol ANR2B為嗎啡+ANR2B組(D組)；在納洛酮激髮前30 min鞘內註射15 nmol ANR2B為戒斷+ANR2B組(E組).應用RT-PCR檢測嗎啡依賴與戒斷大鼠脊髓和腦榦μ受體與κ受體mRNA錶達.結果 ①嗎啡依賴大鼠脊髓和腦榦NR2B mRNA錶達(0.965±0.012；0.958±0.020)較對照組(0.763±0.031)明顯上調,NR2B反義寡覈苷痠使其錶達明顯下調(0.786±0.011).②嗎啡戒斷癥狀主要錶現為濕狗樣搖體、激惹、齒顫、眼瞼下垂、腹瀉等.鞘內註射NR2B反義寡覈苷痠則明顯減輕戒斷癥狀的總評分(32±2,22±3).③嗎啡依賴大鼠脊髓和腦榦中μ受體mRNA錶達(0.955±0.021,0.946±0.013)較對照組升高有統計學意義.鞘內註射ANR2B則明顯降低嗎啡依賴大鼠脊髓和腦榦中μ受體mRNA的錶達(0.801±0.013;0.824±0.021).納洛酮激髮戒斷癥狀後1h,脊髓和腦榦中μ受體mRNA的錶達較嗎啡依賴組明顯下降.激髮前30 min鞘內註射ANR2B,脊髓和腦榦中μ受體mRNA錶達與戒斷組比較變化無統計學意義.④嗎啡依賴大鼠脊髓和腦榦中κ受體mRNA錶達(0.735±0.025；0.757±0.010)較對照組(0.974±0.016；0.982±0.013)降低有統計學意義；同時鞘內註射ANR2B則顯著增加脊髓和腦榦中κ受體mRNA的錶達(0.875±0.012; 0.854±0.016)；納洛酮激髮戒斷癥狀後1h,脊髓和腦榦中κ受體mRNA的錶達較嗎啡依賴組明顯增高；激髮前30min鞘內註射ANR2B,脊髓和腦榦中κ受體mRNA錶達與戒斷組比較變化無統計學意義.各組中β -actin錶達量差異無統計學意義.結論 脊髓和腦榦中μ受體與κ受體錶達的變化在嗎啡依賴與戒斷反應中髮揮瞭重要作用.鞘內註射NR2B反義寡覈苷痠通過有效逆轉嗎啡依賴與戒斷大鼠脊髓和腦榦中μ受體與κ受體錶達的變化趨勢,從而髮揮其抑製嗎啡依賴與減輕戒斷癥狀的作用.
목적 관찰NR2B반의과핵감산(NR2B antisense oligonucleotide,ANR2B)대마배의뢰대서척수화뇌간μ수체여κ수체표체적영향,탐토N-갑기-D-천동안산(N-methyl-D-aspartic acid,NMDA)수체적NR2B아기여아편μ수체여κ수체재개도마배의뢰화내수과정중적상호작용.방법 웅성SD대서,체중230 g～270 g.초내매관성공대서40지,안수궤수자표법분위5조,매조8지.마배피하주사수일10 mg/kg,매일2차,격일매차증가10 mg/kg,지제6천말차주사50 mg/kg,건립마배의뢰모형,이납락동4 mg/kg단차복강주사건립마배의뢰최촉계단모형,관찰납락동최촉계단증상병평개조모시부성공.분위마배의뢰조(A조)；상응시간점피하주사등량생리염수위공백대조조(B조)；말차주사마배후4h복강주사염산납락동4 mg/kg격발계단증상위마배계단조(C조)；재피하주사마배적동시초내주사15 nmol ANR2B위마배+ANR2B조(D조)；재납락동격발전30 min초내주사15 nmol ANR2B위계단+ANR2B조(E조).응용RT-PCR검측마배의뢰여계단대서척수화뇌간μ수체여κ수체mRNA표체.결과 ①마배의뢰대서척수화뇌간NR2B mRNA표체(0.965±0.012；0.958±0.020)교대조조(0.763±0.031)명현상조,NR2B반의과핵감산사기표체명현하조(0.786±0.011).②마배계단증상주요표현위습구양요체、격야、치전、안검하수、복사등.초내주사NR2B반의과핵감산칙명현감경계단증상적총평분(32±2,22±3).③마배의뢰대서척수화뇌간중μ수체mRNA표체(0.955±0.021,0.946±0.013)교대조조승고유통계학의의.초내주사ANR2B칙명현강저마배의뢰대서척수화뇌간중μ수체mRNA적표체(0.801±0.013;0.824±0.021).납락동격발계단증상후1h,척수화뇌간중μ수체mRNA적표체교마배의뢰조명현하강.격발전30 min초내주사ANR2B,척수화뇌간중μ수체mRNA표체여계단조비교변화무통계학의의.④마배의뢰대서척수화뇌간중κ수체mRNA표체(0.735±0.025；0.757±0.010)교대조조(0.974±0.016；0.982±0.013)강저유통계학의의；동시초내주사ANR2B칙현저증가척수화뇌간중κ수체mRNA적표체(0.875±0.012; 0.854±0.016)；납락동격발계단증상후1h,척수화뇌간중κ수체mRNA적표체교마배의뢰조명현증고；격발전30min초내주사ANR2B,척수화뇌간중κ수체mRNA표체여계단조비교변화무통계학의의.각조중β -actin표체량차이무통계학의의.결론 척수화뇌간중μ수체여κ수체표체적변화재마배의뢰여계단반응중발휘료중요작용.초내주사NR2B반의과핵감산통과유효역전마배의뢰여계단대서척수화뇌간중μ수체여κ수체표체적변화추세,종이발휘기억제마배의뢰여감경계단증상적작용.
Objective To observe the effects of NR2B antisense oligonucleotide (ANR2B) on gene expression of μ and κ opiate receptor in spinal cord and brainstem during morphine dependence and withdrawal in rats.To explore the interaction between NR2B subunits of N -methyl-D-aspartic acid (NMDA) receptors,and μ,κ＜ opioid receptors,in the process of mediating morphine dependence and tolerance.Methods Animals were male Sprague Dawley rats weighing 230 g-270 g.Animal models of morphine dependence were established by repeated subcutaneous injection of morphine with progressive doses.Animal models of urged withdrawal in morphine dependence were established by single intraperitoneal injection of naloxone,to morphine dependence rat.After making sure of the successful animal models,40 rats with an intrathecal tube were randomly divided into 5 groups,8 in each group.Group A:the morphine dependence group.Group B:control group.Group C:the morphine withdrawal group.Group D:ANR2B intrathecally injected at the same time of subcutaneous injection of morphine.Group E:intrathecal injection of ANR2B 30mi n before injection of naloxone.The mRNA levels of μ and κ opiate receptor in spinal cord and brainstem were assayed by reverse transcription polymerase chain reaction with the beta-actin mRNA as an internal control.Results Increase in NR2B mRNA expression in the spinal cord and brainstem induced by morphine were significantly attenuated by ANR2B(0.965±0.012,0.958±0.020).The total score of morphine-withdrawal symptoms were significantly decreased by spinal administration of NR2B antisense oligonucleotide (32±2,22±3).The mRNA levels of μ opiate receptor increased significantly in spinal cord and brainstem (0.955±0.021,0.946±0.013)during morphine dependence.But μ opiate receptors mRNA levels decreased after injection of ANR2B (0.801±0.013;0.824±0.021)during morphine withdrawal in rats.The mRNA levels of κ opiate receptor changed conversely compared with the μ opiate receptor during morphine dependence and withdrawal.Spinal administration of NR2B antisense oligonucleotide at the same time of injection of morphine reversed the change tendency of μ and κ opiate receptor in spinal cord and brainstem during morphine dependence and withdrawal in rats.Changes had no statistic significance in spinal administration of NR2B antisense oligonucleotide 30min before injection of naloxone,compared with the withdrawal group.Conclusion The expression of μ and κ opiate receptor in spinal cord and brainstem plays an important role in mediating the development of morphine dependence and withdrawal.NR2B antisense oligonucleotide reversed the expression of μ and κ opiate receptor in spinal cord and brainstem during morphine dependence and withdrawal in rats.It suggests that NR2B antisense oligonucleotide can inhibit the development of morphine dependence and attenuate morphine-withdrawal symptoms by mediating the expression of μ and κ opiate receptors.
