咪多吡對(duì)帕金森病模型大鼠黑質(zhì)紋狀體膠質(zhì)細(xì)胞增生及活化的影響

呂超男,劉 斌,馬原源,苗玉超,劉 穎,張晉霞,毛文靜,孫 靜,成曉華

(河北聯(lián)合大學(xué)附屬醫(yī)院神經(jīng)內(nèi)一科,河北唐山 063000)

咪多吡對(duì)帕金森病模型大鼠黑質(zhì)紋狀體膠質(zhì)細(xì)胞增生及活化的影響

呂超男,劉 斌,馬原源,苗玉超,劉 穎,張晉霞,毛文靜,孫 靜,成曉華

(河北聯(lián)合大學(xué)附屬醫(yī)院神經(jīng)內(nèi)一科,河北唐山 063000)

目的:探討咪多吡對(duì)帕金森病(PD)模型大鼠黑質(zhì)紋狀體膠質(zhì)纖維酸性蛋白(GFAP)和整合素αM (cd11b)表達(dá)的影響,闡明咪多吡對(duì)膠質(zhì)細(xì)胞的調(diào)控作用。方法:72只健康SD大鼠隨機(jī)分為對(duì)照組、PD模型組(模型組)和咪多吡處理組(咪多吡組),每組24只。PD模型組和咪多吡組大鼠采用魚(yú)藤酮制備PD模型,模型制備成功后各組大鼠隨機(jī)分為4 d和8 d 2個(gè)亞組,每個(gè)亞組12只大鼠。免疫組織化學(xué)法和Western blotting法檢測(cè)大鼠黑質(zhì)紋狀體GFAP和cd11b陽(yáng)性細(xì)胞數(shù)和蛋白表達(dá)水平。結(jié)果:對(duì)照組大鼠GFAP和cd11b陽(yáng)性細(xì)胞均處于靜息狀態(tài),GFAP陽(yáng)性細(xì)胞胞體細(xì)長(zhǎng)不規(guī)則,有細(xì)長(zhǎng)的突起,cd11b陽(yáng)性細(xì)胞胞體較小,呈分支狀,突起較細(xì)長(zhǎng);模型組大鼠GFAP和cd11b陽(yáng)性細(xì)胞呈激活狀態(tài),GFAP陽(yáng)性細(xì)胞胞體肥大,突起增多增粗,cd11b陽(yáng)性細(xì)胞胞體增大,突起變粗變短,數(shù)量增多;與模型組比較,咪多吡組大鼠GFAP陽(yáng)性細(xì)胞胞體及突起較細(xì)長(zhǎng), cd11b陽(yáng)性細(xì)胞胞體較小,突起較細(xì)長(zhǎng),數(shù)量減少。對(duì)照組大鼠黑質(zhì)紋狀體可見(jiàn)少量的GFAP和cd11b陽(yáng)性細(xì)胞表達(dá),8 d組與4 d組比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05);模型組大鼠GFAP和cd11b陽(yáng)性細(xì)胞數(shù)和蛋白表達(dá)水平均明顯增加,與對(duì)照組比較差異均有統(tǒng)計(jì)學(xué)意義(P＜0.01),且8 d組多于4 d組,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05);咪多吡組大鼠GFAP和cd11b陽(yáng)性細(xì)胞數(shù)和蛋白表達(dá)水平均明顯降低,與模型組比較差異均有統(tǒng)計(jì)學(xué)意義(P＜0.01),且8 d組少于4 d組,2組比較差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)。結(jié)論:PD模型大鼠黑質(zhì)紋狀體膠質(zhì)細(xì)胞明顯增生和活化,咪多吡能夠抑制膠質(zhì)細(xì)胞的增生和活化。

帕金森病;膠質(zhì)細(xì)胞;膠質(zhì)纖維酸性蛋白;整合素超家族成員Ⅰ型跨膜蛋白;咪多吡

帕金森病(Parkinson’s disease,PD)是一種多因素綜合作用的中樞神經(jīng)系統(tǒng)變性疾病,在遺傳背景、環(huán)境暴露和老齡化的共同作用下,氧化應(yīng)激、線粒體功能衰竭、鈣穩(wěn)態(tài)失衡、興奮性氨基酸毒性和細(xì)胞凋亡等機(jī)制導(dǎo)致黑質(zhì)紋狀體多巴胺能神經(jīng)元大量變性缺失[1]。研究[2]表明:炎癥反應(yīng)在PD的多巴胺(dapamine,DA)能神經(jīng)元死亡的過(guò)程中起著重要作用。膠質(zhì)細(xì)胞是炎癥反應(yīng)的重要參與者,其活化后可產(chǎn)生多種促炎因子,如白細(xì)胞介素1 (IL-1)、白細(xì)胞介素6(IL-6)、巨噬細(xì)胞炎癥蛋白、單核細(xì)胞趨化蛋白1和腫瘤壞死因子α(TNF-α) 等,多種促炎因子損傷了DA能神經(jīng)元,在PD的發(fā)病機(jī)制中起著重要作用[3]。因此,抑制膠質(zhì)細(xì)胞的增生和活化可能成為阻止PD發(fā)展的一條途徑[4]。膠質(zhì)纖維酸性蛋白(glial fibrillary acidic protein,GFAP)的表達(dá)情況可反映星形膠質(zhì)細(xì)胞的增生與活化,是星形膠質(zhì)細(xì)胞的特異性標(biāo)記物[5]。整合素αM(cd11b)對(duì)炎癥細(xì)胞遷移和功能具有重要作用,CD11b的陽(yáng)性表達(dá)能反映小膠質(zhì)細(xì)胞的增生與活化[6],是小膠質(zhì)細(xì)胞的特異性蛋白標(biāo)記物[7]。咪多吡(eldepryl)是一種常用的單胺氧化酶B抑制劑,能保護(hù)黑質(zhì)細(xì)胞免于各種神經(jīng)毒素的侵害而阻止PD進(jìn)展,具有神經(jīng)保護(hù)作用[8],但具體作用機(jī)制尚不明確。本研究采用頸背部皮下注射魚(yú)藤酮制備大鼠PD模型,探討PD模型大鼠黑質(zhì)紋狀體GFAP和cd11b的表達(dá)情況及咪多吡對(duì)其表達(dá)的影響。

1 材料與方法

1.1 實(shí)驗(yàn)動(dòng)物、主要試劑和儀器清潔級(jí)健康雄性SD大鼠,體質(zhì)量250～300 g,購(gòu)于北京華阜康生物科技股份有限公司,合格證號(hào)SCXK(京) 2012-0013,在河北聯(lián)合大學(xué)屏障環(huán)境動(dòng)物實(shí)驗(yàn)室自由進(jìn)食喂養(yǎng),室溫控制在(23±2)℃,自然光照,實(shí)驗(yàn)前適應(yīng)喂養(yǎng)2周。咪多吡(成分:鹽酸司來(lái)吉蘭),批號(hào)H20040400,規(guī)格5 mg/片,芬蘭Orion Corporation Espoo公司產(chǎn)品。魚(yú)藤酮、葵花油、兔抗大鼠GFAP抗體、兔抗大鼠cd11b抗體和免疫組織化學(xué)試劑盒(SP-0023)均購(gòu)自北京博奧森生物工程有限公司,DAB顯色液購(gòu)自北京中杉生物有限公司,SDS-聚丙烯酰胺凝膠電泳低分子量標(biāo)準(zhǔn)蛋白購(gòu)自華美生物公司。低溫離心機(jī)購(gòu)自美國(guó)Sigma公司。

1.2 動(dòng)物分組72只健康SD大鼠隨機(jī)分為對(duì)照組、PD模型組(模型組)和咪多吡處理組(咪多吡組)。每組又分為模型制備成功后4 d和8 d 2個(gè)亞組,每個(gè)亞組各12只大鼠(其中6只用于免疫組織化學(xué)法檢測(cè),6只用于Western blotting法檢測(cè))。

1.3 動(dòng)物模型制備采用頸背部皮下注射魚(yú)藤酮制備大鼠PD模型[9]。具體操作方法:魚(yú)藤酮以葵花油配制成乳液,充分震蕩混勻后避光保存。模型組和咪多吡組大鼠稱體質(zhì)量后以魚(yú)藤酮2 mg·kg－1計(jì)算魚(yú)藤酮葵花油乳液用量。捏起大鼠頸背部皮膚,用1 m L注射器皮下注射魚(yú)藤酮葵花油乳液。對(duì)照組大鼠皮下注射葵花油。將大鼠行為變化分6個(gè)等級(jí)記分[10]:1分,大鼠出現(xiàn)拒捕行為減弱、豎毛、毛色變黃變臟、弓背和主動(dòng)活動(dòng)減少;2分,有1分的表現(xiàn),且主動(dòng)活動(dòng)減少明顯、動(dòng)作遲緩,并有震顫,或有步態(tài)不穩(wěn);4分,有2分的表現(xiàn),且步態(tài)不穩(wěn),或不能直線行走、或行步時(shí)向一側(cè)旋轉(zhuǎn);6分,向單側(cè)斜臥,單側(cè)前肢和(或)后肢癱瘓,行走困難、進(jìn)食困難;8分,單側(cè)前肢和(或)后肢完全癱瘓,四肢拘攣,質(zhì)量大幅度減輕,不能進(jìn)食;10分,瀕死狀態(tài)或死亡。本實(shí)驗(yàn)選取2～6分大鼠入選PD模型。未符合入選標(biāo)準(zhǔn)大鼠隨時(shí)被替換,并重新制備模型大鼠到相應(yīng)組別。本實(shí)驗(yàn)造模后大鼠出現(xiàn)震顫、僵直、運(yùn)動(dòng)減少、活動(dòng)遲緩和行步時(shí)向一側(cè)旋轉(zhuǎn)等行為學(xué)改變,說(shuō)明造模成功。

1.4 給藥方法對(duì)照組:大鼠連續(xù)頸背部皮下注射葵花油2 mg·kg－1,待模型制備成功后停止皮下注射,并灌胃給予生理鹽水。模型組:大鼠連續(xù)頸背部皮下注射魚(yú)藤酮葵花油乳液2 mg·kg－1,待模型制備成功后停止皮下注射,并灌胃給予生理鹽水。咪多吡組:模型制備成功后,每日灌胃給予咪多吡0.5 mg·kg－1,分別連續(xù)給藥4和8 d。

1.5 標(biāo)本制備和檢測(cè)方法免疫組織化學(xué)法:用10%水合氯醛(4 m L·kg－1)腹腔麻醉動(dòng)物,于劍突下剪一約2～3 cm橫切口,沿膈肌與胸廓交界處剪開(kāi)膈肌,向上剪斷3根肋骨,暴露心臟,剪開(kāi)右心耳,經(jīng)心臟快速灌注生理鹽水100～200 m L (4℃)至肝臟完全變白,之后灌入含4%多聚甲醛的0.1 mol·L－1PBS(p H 7.4,4℃)200～400 m L,至大鼠肝臟變硬、肢體僵直,即固定完成。然后完整取出鼠腦置于含4%多聚甲醛的0.1 mol·L－1PBS(p H 7.4,4℃)中過(guò)夜固定保存。參照《大鼠腦立體定位圖譜》[11]在大鼠黑質(zhì)紋狀體取材,進(jìn)行常規(guī)脫水、石蠟包埋。選擇4μm厚度連續(xù)切片,撈片后置于60℃烤箱中烘干0.5 h。取各組制備好的黑質(zhì)紋狀體組織切片,加入正常羊血清封閉液中,分別滴加兔抗大鼠GFAP一抗(1∶200)和兔抗大鼠cd11b一抗(1∶200)4℃過(guò)夜,滴加二抗和適量辣根酶標(biāo)記鏈霉卵白素工作液,進(jìn)行DAB顯色。按試劑盒說(shuō)明書(shū)操作要求檢測(cè)各組大鼠黑質(zhì)紋狀體中GFAP和cd11b蛋白表達(dá)。光鏡下觀察,胞漿呈棕黃色、核呈淺藍(lán)色或紫藍(lán)色為陽(yáng)性細(xì)胞。高倍鏡下觀察陽(yáng)性細(xì)胞形態(tài)變化,并隨機(jī)分別觀察各組大鼠黑質(zhì)紋狀體不重疊的6視野,進(jìn)行GFAP和cd11b陽(yáng)性細(xì)胞計(jì)數(shù)。Western blotting法:用10%水合氯醛(0.3 m L·100 g－1)對(duì)大鼠進(jìn)行深度麻醉。斷頭后立即分離出新鮮黑質(zhì)紋狀體,制取蛋白樣品。取等量的樣品進(jìn)行電泳、轉(zhuǎn)膜、封閉后, 在GFAP蛋白(48 000)和cd11b蛋白(125 000)相對(duì)分子質(zhì)量相應(yīng)位置處剪取PVDF膜,分別加入稀釋好的兔抗大鼠GFAP一抗(1∶1 000)和兔抗大鼠cd11b一抗(1∶250),4℃孵育過(guò)夜,PBS洗膜,

1.6 統(tǒng)計(jì)學(xué)分析采用SPSS 13.0統(tǒng)計(jì)學(xué)軟件進(jìn)行數(shù)據(jù)分析。各組大鼠黑質(zhì)紋狀體中GFAP和CD11b陽(yáng)性細(xì)胞數(shù)及蛋白表達(dá)水平以±s表示,組間比較采用單因素方差分析,2組間均數(shù)比較采用t檢驗(yàn)。

2 結(jié)果

2.1 各組大鼠黑質(zhì)紋狀體中GFAP和cd11b陽(yáng)性細(xì)胞數(shù)對(duì)照組大鼠黑質(zhì)紋狀體可見(jiàn)少量的GFAP 和cd11b陽(yáng)性細(xì)胞表達(dá),GFAP和cd11b陽(yáng)性細(xì)胞均處于靜息狀態(tài),GFAP陽(yáng)性細(xì)胞胞體細(xì)長(zhǎng)不規(guī)則,有細(xì)長(zhǎng)的突起,cd11b陽(yáng)性細(xì)胞胞體較小,呈分支狀,突起較細(xì)長(zhǎng);且GFAP和cd11b陽(yáng)性細(xì)胞數(shù)8 d組與4 d組比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)。模型組大鼠黑質(zhì)紋狀體中GFAP和CD11b陽(yáng)性細(xì)胞數(shù)明顯增多,GFAP和cd11b陽(yáng)性細(xì)胞呈激活狀態(tài),GFAP陽(yáng)性細(xì)胞胞體肥大,突起增多增粗,cd11b陽(yáng)性細(xì)胞胞體增大,突起變粗變短,數(shù)量增多;GFAP和cd11b陽(yáng)性細(xì)胞數(shù)與對(duì)照組比較差異均有統(tǒng)計(jì)學(xué)意義(P＜0.01),且8 d組多于4 d組,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)。咪多吡組黑質(zhì)紋狀體中GFAP和cd11b陽(yáng)性細(xì)胞數(shù)明顯減少,與模型組比較差異均有統(tǒng)計(jì)學(xué)意義(P＜0.01);且GFAP和cd11b陽(yáng)性細(xì)胞數(shù)8 d組少于4 d組,差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)。見(jiàn)表1和圖1～2(插頁(yè)二和三)。

2.2 各組大鼠黑質(zhì)紋狀體中GFAP和cd11b蛋白表達(dá)水平對(duì)照組大鼠黑質(zhì)紋狀體中GFAP和cd11b蛋白表達(dá)水平較低,且8 d組與4 d組比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)。模型組大鼠黑質(zhì)紋狀體中GFAP和cd11b表達(dá)水平明顯增加,與對(duì)照組比較差異均有統(tǒng)計(jì)學(xué)意義(P＜0.01);雖8 d組高于4 d組,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)。咪多吡組大鼠黑質(zhì)紋狀體中GFAP和cd11b蛋白表達(dá)水平明顯降低,與模型組比較差異均有統(tǒng)計(jì)學(xué)意義(P＜0.01);且8 d組低于4 d組, 2組比較差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)。見(jiàn)表2和圖3～4。加人相應(yīng)稀釋好的二抗(羊抗兔,1∶2 000),37℃反應(yīng)1 h,洗膜,以ECL顯色,膠片曝光顯影,將曝光膠片的圖像輸入計(jì)算機(jī),用Image J圖像程序分析軟件分析目的蛋白及內(nèi)參的光密度(A)值,以兩者的比值反映目的蛋白的相對(duì)表達(dá)量。

表1 各組大鼠黑質(zhì)紋狀體中GFAP和cd11b陽(yáng)性細(xì)胞數(shù)Tab.1 Number of cells with positive expression of GFAP and cd11b in substantia nigra and striatum of rats in varions groups (n＝6,±s)

表1 各組大鼠黑質(zhì)紋狀體中GFAP和cd11b陽(yáng)性細(xì)胞數(shù)Tab.1 Number of cells with positive expression of GFAP and cd11b in substantia nigra and striatum of rats in varions groups (n＝6,±s)

?P＜0.05,??P＜0.01 compared with control group;△P＜0.01 compared with model group;＃P＜0.05 compared with 4 d group.

Group GFAP cd11b (t/d) 4 848 Control 13.67±2.94 13.83±2.86 29.50±3.83 30.17±1.94 Model 30.83±3.06??31.67±3.20??62.33±3.07??65.67±4.58??Eldepryl 21.00±2.53??△17.50±2.07?△＃46.50±3.15??△42.00±2.37??△＃

表2 各組大鼠黑質(zhì)紋狀體中GFAP和cd11b蛋白表達(dá)水平Tab.2 Expression levels of GFAP and cd11b in substantia nigra and striatum of rats in varions groups(n＝6,±s)

表2 各組大鼠黑質(zhì)紋狀體中GFAP和cd11b蛋白表達(dá)水平Tab.2 Expression levels of GFAP and cd11b in substantia nigra and striatum of rats in varions groups(n＝6,±s)

?P＜0.01 compared with control group;△P＜0.01 compared with model group;＃P＜0.05 compared with 4 d group.

Group GFAP cd11b (t/d) 4 848 Control 16 755.17±484.60 16 852.67±407.63 15 303.00±637.22 15 948.50±410.87 Model 28 842.83±730.72?29 614.17±1026.23?26 512.67±459.31?27 159.33±649.06?Eldepryl 26 113.17±765.60?△25 304.83±472.87?△＃20 996.50±565.03?△20 323.00±574.94?△＃

圖3 Western blotting法檢測(cè)各組大鼠黑質(zhì)紋狀體中GFAP蛋白表達(dá)電泳圖Fig.3 Electrophoregram of expressins of GFAP in substantia nigra and striatum of rats in varions groups detected by Western blotting methodA:Control group;B:Model group;C:Eldepryl group.Lane 1:4 d;Lane 2:8 d.

圖4 Western blotting法檢測(cè)各組大鼠黑質(zhì)紋狀體中cd11b蛋白表達(dá)電泳圖Fig.4 Electrophoregram of expressions of cd11b in substantia nigra and striatum of rats in varions groups detected by Western blotting methodA:Control group;B:Model group;C:Eldepryl group.Lane 1:4 d;Lane 2:8 d.

3 討論

目前已經(jīng)建立的PD動(dòng)物模型種類較多,采用低劑量頸背部皮下注射魚(yú)藤酮方法制備PD動(dòng)物模型能較好地模擬PD的慢性進(jìn)行性自然病程和發(fā)病特點(diǎn),是目前國(guó)內(nèi)外公認(rèn)的理想的PD動(dòng)物模型之一[12]。

PD主要的病理特征是腦黑質(zhì)DA能神經(jīng)元進(jìn)行性變性喪失,炎癥反應(yīng)在PD的DA能神經(jīng)元死亡的過(guò)程中起著重要作用[2]。膠質(zhì)細(xì)胞是炎癥反應(yīng)的重要參與者,激活的星形膠質(zhì)細(xì)胞釋放一氧化氮(NO)、合成TNF-α、合成興奮性氨基酸等發(fā)揮細(xì)胞毒性作用,造成細(xì)胞損傷,促進(jìn)炎癥反應(yīng)和脫髓鞘,破壞血腦屏障,導(dǎo)致神經(jīng)元的損傷[3]。激活的小膠質(zhì)細(xì)胞產(chǎn)生大量炎性因子和氧自由基發(fā)揮神經(jīng)毒性作用[13],DA能神經(jīng)元對(duì)這些炎癥因子和氧化物具有易感性而造成損傷[14]。死亡的DA能神經(jīng)元又可以促進(jìn)小膠質(zhì)細(xì)胞的活化,如此形成惡性循環(huán),使神經(jīng)退行性病變進(jìn)行性發(fā)展[15]。GFAP是星形膠質(zhì)細(xì)胞的特征性標(biāo)記物,是中樞神經(jīng)系統(tǒng)中星形膠質(zhì)細(xì)胞最常見(jiàn)的特征性反應(yīng)之一[4]。當(dāng)機(jī)體受到某種刺激時(shí),中樞神經(jīng)系統(tǒng)某些部位的星形膠質(zhì)細(xì)胞出現(xiàn)GFAP的陽(yáng)性表達(dá),反映出星形膠質(zhì)細(xì)胞在此時(shí)的功能活動(dòng)增強(qiáng)。cd11b對(duì)于炎癥細(xì)胞遷移和功能具有重要的作用,cd11b是小膠質(zhì)細(xì)胞的特異性蛋白標(biāo)記物[7]。咪多吡是一種選擇性的單胺氧化酶B抑制劑,能增加抗氧化酶的濃度、減輕細(xì)胞凋亡和產(chǎn)生神經(jīng)營(yíng)養(yǎng)因子,保護(hù)黑質(zhì)細(xì)胞免于各種神經(jīng)毒素的侵害[16],在臨床應(yīng)用中顯示出較好的治療效果和耐受性[17]。本研究中免疫組織化學(xué)法和Western blotting法檢測(cè)結(jié)果均顯示:模型組大鼠黑質(zhì)紋狀體中GFAP與cd11b表達(dá)均增多,細(xì)胞呈激活狀態(tài),說(shuō)明PD模型大鼠黑質(zhì)紋狀體有明顯的膠質(zhì)細(xì)胞增生和活化;咪多吡組大鼠黑質(zhì)紋狀體中GFAP與cd11b表達(dá)均減少,說(shuō)明咪多吡能夠抑制膠質(zhì)細(xì)胞的增生和活化,且隨著時(shí)間的延長(zhǎng),作用更明顯。

綜上所述,PD模型大鼠黑質(zhì)紋狀體有明顯的膠質(zhì)細(xì)胞增生和活化,咪多吡能夠抑制膠質(zhì)細(xì)胞的增生和活化,通過(guò)抗炎作用,對(duì)DA能神經(jīng)元產(chǎn)生保護(hù)作用。藥物干預(yù)膠質(zhì)細(xì)胞激活將有助于阻止PD的進(jìn)展。

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Influence of eldepryl in proliferation and activation of gliacytes in substantia nigra and striatum in rats with Parkinson’s disease

LYU Chao-nan,LIU Bin,MA Yuan-yuan,MIAO Yu-chao,LIU Ying,ZHANG Jin-xia, MAO Wen-jing,SUN Jing,CHENG Xiao-hua
(First Department of Neurology,Affiliated Hospital,Hebei United University,Tangshan 063000,China)

ObjectiveTo discuss the influence of eldepryl on the expressions of glial fibrillary acidic protein(GFAP) and cd11b in substantia nigra and striatum in the rats with Parkinson’s disease(PD),and to clarify the regulatory role of eldepryl in the gliacytes.Methods72 SD rats were randomly divided into control group,PD model group and eldepryl group,and each group was divided randomly into 4 d and 8 d subgroups(n＝12)after the success of model preparation.The PD rat models were established by injecting rotenone in subcutaneous.The number of GFAP and cd11b positive cells and the expressions of GFAP and cd11b were detected by immunohistochemistry and Western blotting method.ResultsThe GFAP and cd11b positive cells were all in a resting state in control group, the GFAP-positive cell body was slender and irregular and had elongated protrusions;the cd11b-positive cell body was small and branch-like,and it had more slender protrusions.The GFAP and cd11b positive cells were all in a active state in model group,the GFAP-positive cell body was hypertrophy,the projections increased thickening;the cd11b-positive cell body was more bigger,the projections were shorter and thicker,and the number was increased.Compared with model group,the GFAP-positive cell body and protrusions were more slender,the CD11b-positive cell body was more smaller,the projections were more slender,and the number was decreased in eldepryl group.There were a small amount of expression of GFAP and cd11b positive cells in substantia nigra and striatum in the rats in control group,and there was no significant difference between 8 d group and 4 d group(P＞0.05).The number of GFAP and cd11b positive cells and the protein expression levels were significantly increased in model group compared with control group(P＜0.01);there was more expression in 8 d group compared with 4 d group,but there was no significant difference(P＞0.05).The number of GFAP and cd11b positive cells and the protein expression levels in eldepryl group were significantly reduced compared with model group(P＜0.01);there were less expression in 8 d group compared with 4 d group,and there was significant difference(P＜0.05).ConclusionThere are activation and proliferation of the gliacytes in substantia nigra and striatum in the rats with PD,and eldepryl can inhibit the activation and proliferation of gliacytes.

Parkinson’s disease;gliacytes;glial fibrillary acidic protein;cd11b;eldepryl

R742.5

A

2013-12-29

河北省衛(wèi)生廳醫(yī)學(xué)科學(xué)研究重點(diǎn)項(xiàng)目資助課題(20130064)

呂超男(1986－),女,河北省任丘市人,醫(yī)師,醫(yī)學(xué)碩士,主要從事神經(jīng)變性疾病的基礎(chǔ)與臨床研究。

劉 斌(Tel:0315-3725963,E-mail:liubintsh＠126.com)

1671-587Ⅹ(2014)05-0953-05

10.13481/j.1671-587x.20140509