中国神经免疫学和神经病学杂志
中國神經免疫學和神經病學雜誌
중국신경면역학화신경병학잡지
CHINESE JOURNAL OF NEUROIMMUNOLOGY AND
2014年
6期
414-418
,共5页
张广慧%吴方荣%何明利%郭振委%秦新月
張廣慧%吳方榮%何明利%郭振委%秦新月
장엄혜%오방영%하명리%곽진위%진신월
电刺激%缺血再灌注%神经发生,大鼠
電刺激%缺血再灌註%神經髮生,大鼠
전자격%결혈재관주%신경발생,대서
electrical stimulation%ischemia/reperfusion%neurogenesis,rats
目的：探讨嗅球电刺激对脑缺血再灌注大鼠脑内神经发生的影响。方法将健康雌性SD大鼠随机分为假手术组、脑缺血再灌注组、电刺激组、假刺激组，其中脑缺血再灌注组、电刺激组，以线栓法建立大鼠右侧大脑中动脉缺血（middle cerebral artery occlusion ，MCAO）/再灌注模型，假手术组仅分离出颈内动脉；电刺激组在右侧嗅球内埋置双极电极，于MCAO再灌注后对嗅球进行电刺激，假刺激组只埋置电极，不进行电刺激；以5－溴脱氧尿嘧啶（Brdu）标记内源性神经干细胞（NSC ）。假手术组于术后48 h、1周，脑缺血再灌注组于缺血再灌注后48 h、1周，电刺激组及假刺激组于刺激后48 h、1周分别行Brdu免疫组化染色，观察各组脑室下区（SVZ）区NSC增殖情况；假手术组、缺血再灌注组、电刺激组、假刺激组4组大鼠在电刺激组行电刺激后腹腔注射Brdu 50 mg/kg ，2次/d ，连续2 d ，4周后处死，分别取脑切片，进行免疫组化和免疫荧光双标染色，观察各组NSC迁移和分化情况。结果缺血再灌注组（37.67±1.97）、假刺激组（36.5±2.35）和电刺激组（43.67±1.63）大鼠48 h后SVZ区Brdu阳性NSC细胞数较假手术组（15.5±1.52）明显增多（F＝57.21，P＜0.05），缺血再灌注组（41.17±2.94）、假刺激组（41.83±2.14）和电刺激组（47.67±2.34）1周后Brdu阳性细胞数较假手术组（15.5±1.52）增多（F＝73.62，P＜0.01），电刺激组在各时间点均较其他组Brdu阳性 NSC细胞数增多（P＜0.01）。缺血并注射Brdu 4周后缺血侧皮质区电刺激组Brdu阳性NSC细胞数（57.17±2.4）较缺血再灌注组（46.83±2.48）和假刺激组（45.83±2.14）增多（F＝161.50，P＜0.01）；免疫荧光双标染色示电刺激组Brdu阳性NSC细胞中Brdu/神经元核抗原（Neun ）双阳性细胞比例约为（74.67±3.61）％，Brdu/胶质纤维酸性蛋白（GFAP）双阳性细胞比例约为（38.83±3.36）％，与缺血再灌注组和假刺激组比较差异均无统计学意义（均 P＞0.05）。结论电刺激嗅球可能促进脑缺血大鼠SVZ区NSC增殖，并可能促进新生NSC向缺血皮质区迁移，增加缺血皮质新生神经元数量，但SVZ区新生NSC的分化不受影响。
目的：探討嗅毬電刺激對腦缺血再灌註大鼠腦內神經髮生的影響。方法將健康雌性SD大鼠隨機分為假手術組、腦缺血再灌註組、電刺激組、假刺激組，其中腦缺血再灌註組、電刺激組，以線栓法建立大鼠右側大腦中動脈缺血（middle cerebral artery occlusion ，MCAO）/再灌註模型，假手術組僅分離齣頸內動脈；電刺激組在右側嗅毬內埋置雙極電極，于MCAO再灌註後對嗅毬進行電刺激，假刺激組隻埋置電極，不進行電刺激；以5－溴脫氧尿嘧啶（Brdu）標記內源性神經榦細胞（NSC ）。假手術組于術後48 h、1週，腦缺血再灌註組于缺血再灌註後48 h、1週，電刺激組及假刺激組于刺激後48 h、1週分彆行Brdu免疫組化染色，觀察各組腦室下區（SVZ）區NSC增殖情況；假手術組、缺血再灌註組、電刺激組、假刺激組4組大鼠在電刺激組行電刺激後腹腔註射Brdu 50 mg/kg ，2次/d ，連續2 d ，4週後處死，分彆取腦切片，進行免疫組化和免疫熒光雙標染色，觀察各組NSC遷移和分化情況。結果缺血再灌註組（37.67±1.97）、假刺激組（36.5±2.35）和電刺激組（43.67±1.63）大鼠48 h後SVZ區Brdu暘性NSC細胞數較假手術組（15.5±1.52）明顯增多（F＝57.21，P＜0.05），缺血再灌註組（41.17±2.94）、假刺激組（41.83±2.14）和電刺激組（47.67±2.34）1週後Brdu暘性細胞數較假手術組（15.5±1.52）增多（F＝73.62，P＜0.01），電刺激組在各時間點均較其他組Brdu暘性 NSC細胞數增多（P＜0.01）。缺血併註射Brdu 4週後缺血側皮質區電刺激組Brdu暘性NSC細胞數（57.17±2.4）較缺血再灌註組（46.83±2.48）和假刺激組（45.83±2.14）增多（F＝161.50，P＜0.01）；免疫熒光雙標染色示電刺激組Brdu暘性NSC細胞中Brdu/神經元覈抗原（Neun ）雙暘性細胞比例約為（74.67±3.61）％，Brdu/膠質纖維痠性蛋白（GFAP）雙暘性細胞比例約為（38.83±3.36）％，與缺血再灌註組和假刺激組比較差異均無統計學意義（均 P＞0.05）。結論電刺激嗅毬可能促進腦缺血大鼠SVZ區NSC增殖，併可能促進新生NSC嚮缺血皮質區遷移，增加缺血皮質新生神經元數量，但SVZ區新生NSC的分化不受影響。
목적：탐토후구전자격대뇌결혈재관주대서뇌내신경발생적영향。방법장건강자성SD대서수궤분위가수술조、뇌결혈재관주조、전자격조、가자격조，기중뇌결혈재관주조、전자격조，이선전법건립대서우측대뇌중동맥결혈（middle cerebral artery occlusion ，MCAO）/재관주모형，가수술조부분리출경내동맥；전자격조재우측후구내매치쌍겁전겁，우MCAO재관주후대후구진행전자격，가자격조지매치전겁，불진행전자격；이5－추탈양뇨밀정（Brdu）표기내원성신경간세포（NSC ）。가수술조우술후48 h、1주，뇌결혈재관주조우결혈재관주후48 h、1주，전자격조급가자격조우자격후48 h、1주분별행Brdu면역조화염색，관찰각조뇌실하구（SVZ）구NSC증식정황；가수술조、결혈재관주조、전자격조、가자격조4조대서재전자격조행전자격후복강주사Brdu 50 mg/kg ，2차/d ，련속2 d ，4주후처사，분별취뇌절편，진행면역조화화면역형광쌍표염색，관찰각조NSC천이화분화정황。결과결혈재관주조（37.67±1.97）、가자격조（36.5±2.35）화전자격조（43.67±1.63）대서48 h후SVZ구Brdu양성NSC세포수교가수술조（15.5±1.52）명현증다（F＝57.21，P＜0.05），결혈재관주조（41.17±2.94）、가자격조（41.83±2.14）화전자격조（47.67±2.34）1주후Brdu양성세포수교가수술조（15.5±1.52）증다（F＝73.62，P＜0.01），전자격조재각시간점균교기타조Brdu양성 NSC세포수증다（P＜0.01）。결혈병주사Brdu 4주후결혈측피질구전자격조Brdu양성NSC세포수（57.17±2.4）교결혈재관주조（46.83±2.48）화가자격조（45.83±2.14）증다（F＝161.50，P＜0.01）；면역형광쌍표염색시전자격조Brdu양성NSC세포중Brdu/신경원핵항원（Neun ）쌍양성세포비례약위（74.67±3.61）％，Brdu/효질섬유산성단백（GFAP）쌍양성세포비례약위（38.83±3.36）％，여결혈재관주조화가자격조비교차이균무통계학의의（균 P＞0.05）。결론전자격후구가능촉진뇌결혈대서SVZ구NSC증식，병가능촉진신생NSC향결혈피질구천이，증가결혈피질신생신경원수량，단SVZ구신생NSC적분화불수영향。
Objective To investigate the effect of olfactory bulb (OB ) electrical stimulation on neurogenesis of rats suffered from ischemia/reperfusion injury .Methods Healthy adult female Sprague‐Dawley (SD) rats were randomly divided into the sham‐operation group ,the ischemia/reperfusion (I/R) group ,the stimulation group ,and the sham‐stimulation group .Cerebral ischemia/reperfusion in the I/R group and the stimulation/sham‐stimulation group was induced by intraluminal right middle cerebral artery occlusion (MCAO) . Arteries in the sham‐operation group were dissected but not occluded .Electrical stimulation was performed via a bipolar electrode implanted in the right OB of rats , and rats in the sham‐stimulation group were implanted electrodes but not stimulated .Bromodeoxyuridine (Brdu) was injected intraperitoneally to label endogenous neural stem cells (NSC ) . The animals in all groups were sacrificed at 48 h , 1 week after corresponding intervention .Immunohistochemistry staining was used to detect the proliferation of NSC in subventricular zone (SVZ) .After injection with Brdu ,the rats were sacrificed 4 weeks after stimulation .Fluorescent double staining for Brdu/Neuronal nuclei antigen (Neun) and Brdu/Glial fibrillary acidic protein (GFAP) was performed in the brain slices finally .Results In SVZ of rats ,Brdu‐positive cells began to increase 48 h after ischemia in the I/R group and the stimulation group .There were more Brdu positive cells in the stimulation group than the I/R group at 48 h (43.67 ± 1.63 , 37.67 ± 1.97; P< 0.05 ) and 7 d (47.67 ± 2.34 , 41.17 ± 2.94 ;P< 0.01 ) af ter ischemia/reperfusion in rats .Four weeks after stimulation ,the Brdu‐positive cells were significantly increased in cortex of the stimulation group (57.17 ± 2.4) than I/R group (46.83 ± 2.48) and sham‐operation group (45.83 ± 2.14 ) ( P < 0.01 ) . Fluorescence double staining showed that stimulation of OB had no effect on the differentiation of NSC into neurons or gliocytes (P>0.05) .Conclusions These observations demonstrated that electrical stimulation of OB might enhance proliferation of NSC in SVZ and promote migration of NSC to ischemic cortex ,but OB stimulation has no effect on differentiation of NSC .
