中华神经科杂志
中華神經科雜誌
중화신경과잡지
Chinese Journal of Neurology
2011年
12期
820-825
,共6页
杨新新%任甜甜%吴娜%宋璐%袁伟恩%刘振国
楊新新%任甜甜%吳娜%宋璐%袁偉恩%劉振國
양신신%임첨첨%오나%송로%원위은%류진국
帕金森病%运动失调%左旋多巴%苄丝肼%微球体
帕金森病%運動失調%左鏇多巴%芐絲肼%微毬體
파금삼병%운동실조%좌선다파%변사정%미구체
Parkinson disease%Dyskinesias%Levodopa%Benserazide%Microspheres
目的 观察聚乳酸-羟基乙酸共聚物( PLGA)包裹的可释放左旋多巴和苄丝肼的微球对帕金森病(PD)大鼠运动症状及异动症发生的影响并探讨其机制.方法 PLGA包裹左旋多巴及苄丝肼制作微球,高效液相法测定微球在体内释放出的左旋多巴和苄丝肼的浓度,6-羟基多巴胺(6-OHDA)注射制作PD大鼠模型,制模成功的PD大鼠随机分成PD组、左旋多巴组、微球组(每组12只),另设溶剂注射假手术组(n=12).左旋多巴组和微球组大鼠分别接受左旋多巴和苄丝肼(左旋多巴12 mg/kg,苄丝肼15 mg/kg)或等剂量微球皮下注射,在治疗的第1、4、7、10、14天行大鼠前肢功能测定,治疗2周后行大鼠异常不自主运动( AIM)评分,免疫组织化学及Western blot法检测纹状体区磷酸化的多巴胺和环磷腺苷调节的磷酸化蛋白-32(DARPP-32)(Thr34)和△FosB水平.结果 体内释放实验表明第7天时左旋多巴和苄丝肼释放量分别达76.2％和83.6％.微球处理组大鼠在治疗的第10天和第14天时前肢跨步数分别为5.8±1.6和5.2±1.5,比左旋多巴组(2.4±1.1、1.2±0.5)明显增加(t =4.12,5.43,均P＜0.01).微球处理组大鼠第14天AIM评分[(16.0±2.1)分]较左旋多巴处理组[(26.0±3.2)分]显著下降,差异有统计学意义(t =6.59,P＜0.01).免疫组织化学显示微球处理组大鼠纹状体磷酸化DARPP-32水平[(3.7±1.3)×104]较左旋多巴处理组[(7.9±2.2)×104]明显降低(t=2.95,P＜0.05).Western blot结果显示微球处理组大鼠磷酸化DARPP-32和△FosB水平分别为119.4％±11.3％和149.3％±12.3％,较左旋多巴组(184.8％±13.7％和300.4％±14.2％)显著下降(t =4.12、2.91,均P＜0.05).结论 微球皮下注射可以改善PD大鼠的运动症状,同时可以减少PD大鼠异动症的发生,这与微球释放的左旋多巴持续性刺激PD大鼠从而减少磷酸化DARPP-32和△FosB的水平有关.
目的 觀察聚乳痠-羥基乙痠共聚物( PLGA)包裹的可釋放左鏇多巴和芐絲肼的微毬對帕金森病(PD)大鼠運動癥狀及異動癥髮生的影響併探討其機製.方法 PLGA包裹左鏇多巴及芐絲肼製作微毬,高效液相法測定微毬在體內釋放齣的左鏇多巴和芐絲肼的濃度,6-羥基多巴胺(6-OHDA)註射製作PD大鼠模型,製模成功的PD大鼠隨機分成PD組、左鏇多巴組、微毬組(每組12隻),另設溶劑註射假手術組(n=12).左鏇多巴組和微毬組大鼠分彆接受左鏇多巴和芐絲肼(左鏇多巴12 mg/kg,芐絲肼15 mg/kg)或等劑量微毬皮下註射,在治療的第1、4、7、10、14天行大鼠前肢功能測定,治療2週後行大鼠異常不自主運動( AIM)評分,免疫組織化學及Western blot法檢測紋狀體區燐痠化的多巴胺和環燐腺苷調節的燐痠化蛋白-32(DARPP-32)(Thr34)和△FosB水平.結果 體內釋放實驗錶明第7天時左鏇多巴和芐絲肼釋放量分彆達76.2％和83.6％.微毬處理組大鼠在治療的第10天和第14天時前肢跨步數分彆為5.8±1.6和5.2±1.5,比左鏇多巴組(2.4±1.1、1.2±0.5)明顯增加(t =4.12,5.43,均P＜0.01).微毬處理組大鼠第14天AIM評分[(16.0±2.1)分]較左鏇多巴處理組[(26.0±3.2)分]顯著下降,差異有統計學意義(t =6.59,P＜0.01).免疫組織化學顯示微毬處理組大鼠紋狀體燐痠化DARPP-32水平[(3.7±1.3)×104]較左鏇多巴處理組[(7.9±2.2)×104]明顯降低(t=2.95,P＜0.05).Western blot結果顯示微毬處理組大鼠燐痠化DARPP-32和△FosB水平分彆為119.4％±11.3％和149.3％±12.3％,較左鏇多巴組(184.8％±13.7％和300.4％±14.2％)顯著下降(t =4.12、2.91,均P＜0.05).結論 微毬皮下註射可以改善PD大鼠的運動癥狀,同時可以減少PD大鼠異動癥的髮生,這與微毬釋放的左鏇多巴持續性刺激PD大鼠從而減少燐痠化DARPP-32和△FosB的水平有關.
목적 관찰취유산-간기을산공취물( PLGA)포과적가석방좌선다파화변사정적미구대파금삼병(PD)대서운동증상급이동증발생적영향병탐토기궤제.방법 PLGA포과좌선다파급변사정제작미구,고효액상법측정미구재체내석방출적좌선다파화변사정적농도,6-간기다파알(6-OHDA)주사제작PD대서모형,제모성공적PD대서수궤분성PD조、좌선다파조、미구조(매조12지),령설용제주사가수술조(n=12).좌선다파조화미구조대서분별접수좌선다파화변사정(좌선다파12 mg/kg,변사정15 mg/kg)혹등제량미구피하주사,재치료적제1、4、7、10、14천행대서전지공능측정,치료2주후행대서이상불자주운동( AIM)평분,면역조직화학급Western blot법검측문상체구린산화적다파알화배린선감조절적린산화단백-32(DARPP-32)(Thr34)화△FosB수평.결과 체내석방실험표명제7천시좌선다파화변사정석방량분별체76.2％화83.6％.미구처리조대서재치료적제10천화제14천시전지과보수분별위5.8±1.6화5.2±1.5,비좌선다파조(2.4±1.1、1.2±0.5)명현증가(t =4.12,5.43,균P＜0.01).미구처리조대서제14천AIM평분[(16.0±2.1)분]교좌선다파처리조[(26.0±3.2)분]현저하강,차이유통계학의의(t =6.59,P＜0.01).면역조직화학현시미구처리조대서문상체린산화DARPP-32수평[(3.7±1.3)×104]교좌선다파처리조[(7.9±2.2)×104]명현강저(t=2.95,P＜0.05).Western blot결과현시미구처리조대서린산화DARPP-32화△FosB수평분별위119.4％±11.3％화149.3％±12.3％,교좌선다파조(184.8％±13.7％화300.4％±14.2％)현저하강(t =4.12、2.91,균P＜0.05).결론 미구피하주사가이개선PD대서적운동증상,동시가이감소PD대서이동증적발생,저여미구석방적좌선다파지속성자격PD대서종이감소린산화DARPP-32화△FosB적수평유관.
Objective To investigate the effects of levodopa/benserazide-loaded poly-lactide-coglycolide (PLGA) microspheres on motor deficits and levodopa-induced dyskinesia in a rat model of Parkinson' s disease (PD) and to explore the mechanisms underlying this effects.Methods The content of levodopa/benserazide released from the microspheres was determined by high-performance liquid chromatography.The rat model of PD was induced by 6-OHDA injections.Then the valid PD rats were treated with levodopa ( 12 mg/kg,s.c.)/benserazide ( 15 mg/kg,s.c.) or microspheres.Forepaw adjusting steps were measured on 1,4,7,10 and 14 days after treatment.After 2 weeks of treatment,the abnormal involuntary movements (AIM) were measured.Phosphorylated dopamine,cAMP-regulated phosphoprotein of 32 kDa (DARPP-32) at threonine 34 levels were determined by immunohistochemistry and Western blot respectively.In addition,the levels of △FosB were measured by Western blot.Results In vivo release test showed that 76.2％ of levodopa and 83.6％ benserazide were released from the microspheres on day 7.Forepaw adjusting steps showed that the scores of forepaw adjusting steps in microspheres-treated PD rats were ( 5.8 ± 1.6 ) and ( 5.2 ± 1.5 ) respectively on 10 and 14 days of treatment,which were increased compared to levodopa-treaded PD rats (2.4 ± 1.1 and 1.2 ± 0.5 ; t =4.12,5.43,all P ＜0.01 ).The AIM scores of microspheres-treated rats ( 16.0 ±2.1 ) were decreased significantly compared to levodopa-treated rats ( 26.0 ± 3.2 ) on day 14 ( t =6.59,P ＜ 0.01 ).Immunohistochemistry indicated that the phosphorylated levels of DARPP-32 in microspheres-treated rats ( (3.7 ± 1.3 ) × 104 )were decreased significantly compared to levodopa-treated rats ( (7.9 ± 2.2) × 104 ; t =2.95,P ＜ 0.05 ).In addition,Western blot showed that the levels of phosphorylated DARPP-32 and △FosB were 119.4％ ± 11.3％ and 149.3％ ± 12.3％ respectively,which were decreased significantly compared to levodopatreated rats ( 184.8％ ± 13.7％ and 300.4％ ± 14.2％ ; t =4.12,2.91,all P ＜ 0.05 ).Conclusions Microspheres can be used to improve the motor deficits and reduce the expression of dyskinesia in PD rats.This may be due to the continuous release of levodopa/beserazide from the microspheres,which leads to continuous stimulation of PD rats and reduces the levels of phosphorylated DARPP-32 and △FosB in the striata of these rats.
