中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2010年
1期
39-44
,共6页
何志江%曹洁%蔡方成%冯成功%陈恒胜
何誌江%曹潔%蔡方成%馮成功%陳恆勝
하지강%조길%채방성%풍성공%진항성
癫(癎)%肌阵挛性%脑干%疾病模型%动物
癲(癎)%肌陣攣性%腦榦%疾病模型%動物
전(간)%기진련성%뇌간%질병모형%동물
Epilepsies%myoclonic%Brain stem%Disease models%animal
目的 探讨建立在发生机制、发作行为、神经电生理学及药效学特征等方面均与临床脑干起源肌阵挛更为一致的实验动物模型.方法 以5-羟色胺(5-HT)的前体L-5-羟色胺酸(L-5-HTP)在健康幼年豚鼠脑桥背侧定点微量注射诱发肌阵挛(同步记录的脑电图暴发活动≤400 ms),观察肌阵挛发作潜伏期、达峰时间、最大发作频率、高峰持续时间和总持续时间等行为学特征.以多导电生理同步记录肌阵挛发作期脑电图、肌电图以及抽动逆向锁定的脑电叠加分析(JLA),论证及认定肌阵挛为脑干起源,并选择对控制肌阵挛具不同效力的丙戊酸(VPA)、氯硝西泮(CZP)和卡马西平(CBZ)等抗癫(癎)药物(AEDs),按达到半数有效浓度(EC_(50))的剂量预处理实验动物后,根据各自药效学特征择时诱导肌阵挛发作,观测抗癫(癎)药物预处理后肌阵挛发作行为及电生理学的变化特征.结果 (1)在一定剂量范围内,L-5-HTP在8只豚鼠脑桥背侧部微量注射,全部一次性诱发单纯性肌阵挛发作,诱发成功率100%.(2)行为学特征:脑桥起源肌阵挛发作表现为两侧或全身性肌阵挛,并显示对触摸、声音刺激的敏感性.(3)脑桥起源肌阵挛肌电暴发时程较长,达(208.75±81.42)ms,同步脑电图中,有散在不规则棘、尖波发放,但与肌电图活动不存在锁时关联.(4)脑桥起源肌阵挛发作中脑电图虽然时有散在棘、尖波发放,但经JLA叠加后无锁时关联的脑电图叠加波.(5)在一次性给药后达到EC_(50)浓度下,VPA和CZP使脑桥起源肌阵挛的最大发作频率[VPA组(28.13±3.79)次/min;CZP组(37.17±4.67)次/min]较对照组[(56.25±6.96)次/min]减少、高峰持续时间[VPA组(55.00±14.14)min;CZP组(50.00±11.73)min]和总持续时间[VPA组(124.17±40.04)min;CZP组(156.88±30.71)min]较对照组[高蜂持续时间(80.00±16.01)min;总持续时间(218.75±17.63)min]显著缩短(F=23.41～35.44,P＜0.01或P＜0.05),对肌阵挛的肌电图时程未有显著影响;CBZ使肌阵挛发作的高峰持续时间[(98.75±13.86)min]和总持续时间[(257.50±14.79)min]较对照组明显延长(P＜0.05、0.01).结论 本模型不仅保证了脑干肌阵挛纯起源,同时因使模型肌阵挛暴发时程显著缩短至400 ms以内,对触觉敏感和对CZP治疗显著有效等优点,较既往L-5-HTP全身注射造模更接近临床实际,从而成功获得在发作行为、神经电生理及药效学特性均与临床更趋一致的脑干起源肌阵挛模型.
目的 探討建立在髮生機製、髮作行為、神經電生理學及藥效學特徵等方麵均與臨床腦榦起源肌陣攣更為一緻的實驗動物模型.方法 以5-羥色胺(5-HT)的前體L-5-羥色胺痠(L-5-HTP)在健康幼年豚鼠腦橋揹側定點微量註射誘髮肌陣攣(同步記錄的腦電圖暴髮活動≤400 ms),觀察肌陣攣髮作潛伏期、達峰時間、最大髮作頻率、高峰持續時間和總持續時間等行為學特徵.以多導電生理同步記錄肌陣攣髮作期腦電圖、肌電圖以及抽動逆嚮鎖定的腦電疊加分析(JLA),論證及認定肌陣攣為腦榦起源,併選擇對控製肌陣攣具不同效力的丙戊痠(VPA)、氯硝西泮(CZP)和卡馬西平(CBZ)等抗癲(癎)藥物(AEDs),按達到半數有效濃度(EC_(50))的劑量預處理實驗動物後,根據各自藥效學特徵擇時誘導肌陣攣髮作,觀測抗癲(癎)藥物預處理後肌陣攣髮作行為及電生理學的變化特徵.結果 (1)在一定劑量範圍內,L-5-HTP在8隻豚鼠腦橋揹側部微量註射,全部一次性誘髮單純性肌陣攣髮作,誘髮成功率100%.(2)行為學特徵:腦橋起源肌陣攣髮作錶現為兩側或全身性肌陣攣,併顯示對觸摸、聲音刺激的敏感性.(3)腦橋起源肌陣攣肌電暴髮時程較長,達(208.75±81.42)ms,同步腦電圖中,有散在不規則棘、尖波髮放,但與肌電圖活動不存在鎖時關聯.(4)腦橋起源肌陣攣髮作中腦電圖雖然時有散在棘、尖波髮放,但經JLA疊加後無鎖時關聯的腦電圖疊加波.(5)在一次性給藥後達到EC_(50)濃度下,VPA和CZP使腦橋起源肌陣攣的最大髮作頻率[VPA組(28.13±3.79)次/min;CZP組(37.17±4.67)次/min]較對照組[(56.25±6.96)次/min]減少、高峰持續時間[VPA組(55.00±14.14)min;CZP組(50.00±11.73)min]和總持續時間[VPA組(124.17±40.04)min;CZP組(156.88±30.71)min]較對照組[高蜂持續時間(80.00±16.01)min;總持續時間(218.75±17.63)min]顯著縮短(F=23.41～35.44,P＜0.01或P＜0.05),對肌陣攣的肌電圖時程未有顯著影響;CBZ使肌陣攣髮作的高峰持續時間[(98.75±13.86)min]和總持續時間[(257.50±14.79)min]較對照組明顯延長(P＜0.05、0.01).結論 本模型不僅保證瞭腦榦肌陣攣純起源,同時因使模型肌陣攣暴髮時程顯著縮短至400 ms以內,對觸覺敏感和對CZP治療顯著有效等優點,較既往L-5-HTP全身註射造模更接近臨床實際,從而成功穫得在髮作行為、神經電生理及藥效學特性均與臨床更趨一緻的腦榦起源肌陣攣模型.
목적 탐토건립재발생궤제、발작행위、신경전생이학급약효학특정등방면균여림상뇌간기원기진련경위일치적실험동물모형.방법 이5-간색알(5-HT)적전체L-5-간색알산(L-5-HTP)재건강유년돈서뇌교배측정점미량주사유발기진련(동보기록적뇌전도폭발활동≤400 ms),관찰기진련발작잠복기、체봉시간、최대발작빈솔、고봉지속시간화총지속시간등행위학특정.이다도전생리동보기록기진련발작기뇌전도、기전도이급추동역향쇄정적뇌전첩가분석(JLA),론증급인정기진련위뇌간기원,병선택대공제기진련구불동효력적병무산(VPA)、록초서반(CZP)화잡마서평(CBZ)등항전(간)약물(AEDs),안체도반수유효농도(EC_(50))적제량예처리실험동물후,근거각자약효학특정택시유도기진련발작,관측항전(간)약물예처리후기진련발작행위급전생이학적변화특정.결과 (1)재일정제량범위내,L-5-HTP재8지돈서뇌교배측부미량주사,전부일차성유발단순성기진련발작,유발성공솔100%.(2)행위학특정:뇌교기원기진련발작표현위량측혹전신성기진련,병현시대촉모、성음자격적민감성.(3)뇌교기원기진련기전폭발시정교장,체(208.75±81.42)ms,동보뇌전도중,유산재불규칙극、첨파발방,단여기전도활동불존재쇄시관련.(4)뇌교기원기진련발작중뇌전도수연시유산재극、첨파발방,단경JLA첩가후무쇄시관련적뇌전도첩가파.(5)재일차성급약후체도EC_(50)농도하,VPA화CZP사뇌교기원기진련적최대발작빈솔[VPA조(28.13±3.79)차/min;CZP조(37.17±4.67)차/min]교대조조[(56.25±6.96)차/min]감소、고봉지속시간[VPA조(55.00±14.14)min;CZP조(50.00±11.73)min]화총지속시간[VPA조(124.17±40.04)min;CZP조(156.88±30.71)min]교대조조[고봉지속시간(80.00±16.01)min;총지속시간(218.75±17.63)min]현저축단(F=23.41～35.44,P＜0.01혹P＜0.05),대기진련적기전도시정미유현저영향;CBZ사기진련발작적고봉지속시간[(98.75±13.86)min]화총지속시간[(257.50±14.79)min]교대조조명현연장(P＜0.05、0.01).결론 본모형불부보증료뇌간기진련순기원,동시인사모형기진련폭발시정현저축단지400 ms이내,대촉각민감화대CZP치료현저유효등우점,교기왕L-5-HTP전신주사조모경접근림상실제,종이성공획득재발작행위、신경전생리급약효학특성균여림상경추일치적뇌간기원기진련모형.
Objective To develop experimental animal model of the brain stem myoclonus,which more closely replicate clinic features of mechanism, behavior, neuroelectrophysiology and pharmacodynamics.Methods L-5-HTP (the precursor of L-5-HT)was microinjected into the dorsal pons of young guinea pig to induce myoclonus (electromyogram burst of myoclonus≤400 ms by synchronous recording).Some animals were pretreated with anticonvulsant VPA,CZP or CBZ at effective dose 50 (EC_(50)).Myoclonus was induced when the drug level was within their effective anticonvulsion concentration.The neuroelectrophysiological characteristics of myoclonus including latency,time of reaching its peak,duration of seizure peak,the maximum seizure frequency and total duration were detected.EMG and ictal electroencephalogram(EEG)were recorded synchronously.The origin of myoclonus and its correlation with epileptic discharges were further confirmed by jerk-locked back averaging(JLA).Results (1)L-5-HTP induced pure myoclonus from the dorsal pons of guinea pig permanently(8/every site,the rate of producing myoclonus is 100%).(2)The myoclonus presented bilaterally or as general myoclonus,which was sensitive to tactile and sound sensation.(3)The EMG duration of the myoclonus wag longer((208.75 ± 81.42)ms),and ictal EEG showed scattered and irregular spikes and sharp waves without time-locked correlation with EMG activities.(4)The synchronous ictal EEG of the myoclonus showed spike and sharp waves,but there was no time-locked EEG activity in JLA.(5)In the animals treated with anticonvulsant at EC_(50) concentrations:VPA and CZP decreased the maximum seizure frequency(there are 28.13±3.79 per minutes in VPA group and 37.17±4.67 perminutes in CZP group)and shortened the duration of peak time ((55.00±14.14)minutes in VPA group and(50.00±11.73)minutes in CZP group respectively)and total time(VPA group was(124.17±40.04)minutes and CZP group was(156.88±30.71)minutes)of myoclonus(F value were between 23.41 and 35.44,P＜0.01 or P＜0.05).CBZ increased duration of peak time((98.75±13.86)minutes)and total time((257.50±14.79)minutes)of myoclonus(P＜0.05 and 0.01).Conclusions The new model generates pure myoclonus originating from brain stem and also has a shorter duration of muscle construction(≤400 ms)and more sensitivity to tactile and sound sensation.Therefore,the model presents characteristics closer to the brain stem myoclonus in the clinic phenotype in respect of seizure behavior,pharmacodynamics and neuroelectrophysiology.
