中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
Chinese Journal of Clinicians(Electronic Edition)
2015年
18期
3404-3408
,共5页
再灌注损伤%血脑屏障%集落刺激因子,重组%水通道蛋白4%胶质纤维酸性蛋
再灌註損傷%血腦屏障%集落刺激因子,重組%水通道蛋白4%膠質纖維痠性蛋
재관주손상%혈뇌병장%집락자격인자,중조%수통도단백4%효질섬유산성단
Reperfusion injury%Blood-brain barrier%Colony-stimulating factors%recombinant%Aquaporin 4%Glail fibrillary acidic protein
目的 探讨重组人粒细胞集落刺激因子(rhG-CSF,瑞白,150 μg,山东齐鲁药业)对SD大鼠脑缺血再灌注后血脑屏障的保护机制.方法 将雄性SD大鼠48只随机分为:假手术组、模型组和治疗组,每组16只,利用改良Longa线栓法制作大鼠中动脉缺血模型,治疗组于缺血2 h后实施再灌注并给予rhG-CSF(50 μg/kg)腹部皮下注射,假手术组及模型组给予同等剂量生理盐水.采用Longa 5分制评分标准进行神经功能评分,利用分光光度仪计算缺血大脑半球脑组织内中伊文思蓝含量,免疫组化法检测各组大鼠脑组织中水通道蛋白-4(AQP4)、胶质纤维酸性蛋白(GFAP)含量变化和电镜下观察血脑屏障的超微结构.结果 治疗组神经功能评分(1.36±0.63)低于模型组(2.50±0.65)(U=16,P<0.01);与假手术组右侧脑组织内伊文思蓝含量(7.38±2.71)相比较,模型组右侧伊文思蓝的含量(37.15±2.30)明显增多(t=30.60,P<0.01),治疗组伊文思蓝的含量(22.75±4.61)较模型组降低(t=-16.73,P<0.05);治疗组AQP4、GFAP表达的灰度值(180.67±7.72、160.64±5.07)均较模型组增高(t=24.16,P<0.01;t=17.98,P<0.01);假手术组大鼠脑组织AQP4、GFAP 表达的灰度值(202.08±5.80、173.73±4.40)高于模型组(t=46.07,P<0.01;t=31.07,P<0.01),电镜下观察到假手术组内皮细胞紧密连接完整,血脑屏障周围组织完好,而模型组和治疗组脑血管内皮细胞连接间隙增加,且连接内皮细胞的基质和基膜完整性缺失.结论 rhG-CSF对脑缺血再灌注损伤具有保护作用,推测其可能机制为rhG-CSF通过抑制星形胶质细胞过度活化,下调GFAP、AQP4的表达,减轻脑水肿,从而保护血脑屏障完整性.
目的 探討重組人粒細胞集落刺激因子(rhG-CSF,瑞白,150 μg,山東齊魯藥業)對SD大鼠腦缺血再灌註後血腦屏障的保護機製.方法 將雄性SD大鼠48隻隨機分為:假手術組、模型組和治療組,每組16隻,利用改良Longa線栓法製作大鼠中動脈缺血模型,治療組于缺血2 h後實施再灌註併給予rhG-CSF(50 μg/kg)腹部皮下註射,假手術組及模型組給予同等劑量生理鹽水.採用Longa 5分製評分標準進行神經功能評分,利用分光光度儀計算缺血大腦半毬腦組織內中伊文思藍含量,免疫組化法檢測各組大鼠腦組織中水通道蛋白-4(AQP4)、膠質纖維痠性蛋白(GFAP)含量變化和電鏡下觀察血腦屏障的超微結構.結果 治療組神經功能評分(1.36±0.63)低于模型組(2.50±0.65)(U=16,P<0.01);與假手術組右側腦組織內伊文思藍含量(7.38±2.71)相比較,模型組右側伊文思藍的含量(37.15±2.30)明顯增多(t=30.60,P<0.01),治療組伊文思藍的含量(22.75±4.61)較模型組降低(t=-16.73,P<0.05);治療組AQP4、GFAP錶達的灰度值(180.67±7.72、160.64±5.07)均較模型組增高(t=24.16,P<0.01;t=17.98,P<0.01);假手術組大鼠腦組織AQP4、GFAP 錶達的灰度值(202.08±5.80、173.73±4.40)高于模型組(t=46.07,P<0.01;t=31.07,P<0.01),電鏡下觀察到假手術組內皮細胞緊密連接完整,血腦屏障週圍組織完好,而模型組和治療組腦血管內皮細胞連接間隙增加,且連接內皮細胞的基質和基膜完整性缺失.結論 rhG-CSF對腦缺血再灌註損傷具有保護作用,推測其可能機製為rhG-CSF通過抑製星形膠質細胞過度活化,下調GFAP、AQP4的錶達,減輕腦水腫,從而保護血腦屏障完整性.
목적 탐토중조인립세포집락자격인자(rhG-CSF,서백,150 μg,산동제로약업)대SD대서뇌결혈재관주후혈뇌병장적보호궤제.방법 장웅성SD대서48지수궤분위:가수술조、모형조화치료조,매조16지,이용개량Longa선전법제작대서중동맥결혈모형,치료조우결혈2 h후실시재관주병급여rhG-CSF(50 μg/kg)복부피하주사,가수술조급모형조급여동등제량생리염수.채용Longa 5분제평분표준진행신경공능평분,이용분광광도의계산결혈대뇌반구뇌조직내중이문사람함량,면역조화법검측각조대서뇌조직중수통도단백-4(AQP4)、효질섬유산성단백(GFAP)함량변화화전경하관찰혈뇌병장적초미결구.결과 치료조신경공능평분(1.36±0.63)저우모형조(2.50±0.65)(U=16,P<0.01);여가수술조우측뇌조직내이문사람함량(7.38±2.71)상비교,모형조우측이문사람적함량(37.15±2.30)명현증다(t=30.60,P<0.01),치료조이문사람적함량(22.75±4.61)교모형조강저(t=-16.73,P<0.05);치료조AQP4、GFAP표체적회도치(180.67±7.72、160.64±5.07)균교모형조증고(t=24.16,P<0.01;t=17.98,P<0.01);가수술조대서뇌조직AQP4、GFAP 표체적회도치(202.08±5.80、173.73±4.40)고우모형조(t=46.07,P<0.01;t=31.07,P<0.01),전경하관찰도가수술조내피세포긴밀련접완정,혈뇌병장주위조직완호,이모형조화치료조뇌혈관내피세포련접간극증가,차련접내피세포적기질화기막완정성결실.결론 rhG-CSF대뇌결혈재관주손상구유보호작용,추측기가능궤제위rhG-CSF통과억제성형효질세포과도활화,하조GFAP、AQP4적표체,감경뇌수종,종이보호혈뇌병장완정성.
Objective To explore the protection mechanism of recombinant human granulocyte colony stimulating factor (rhG-CSF) on blood brain barrier (BBB) after damage following cerebral ischemia-reperfusion.Methods Forty-eight male SD rats were randomly divided into the Sham-operated group, the model group and the rhG-CSF treatment group, 16 rats were included in each group. The middle cerebral artery occlusion was established by the modified Longa suture method. In the treatment group, a single dose of 50 μg/kg rhG-CSF was injected subcutaneously after cerebral ischemia 2 hours. The other groups were given the same volume of saline. Using Longa 5-point scale to estimate the neurological function; The expression level of AQP4 and GFAP were detected by immunohisto-chemistry; The ultra structure changes of blood brain barrier after cerebral ischemic reperfusion were observed by transmission electron microscopic technology.Results The treatment group neurological function score (1.36±0.63) was lower than the control model group (2.50±0.65) (U=16,P<0.05); Comparing with the Sham-operated group (7.38±2.71), the control model group of Evans blue content (37.15±2.30) significantly increased(t=30.60,P<0.01), the treatment group of Evans blue content (22.75±4.61) compared with the control model group decreased (t=-16.73,P<0.01); The treatment group AQP4, GFAP expression of grey value, respectively 180.67±7.72, 160.64±5.07, were higher than the control model group (t=24.16,P<0.01, t=17.98,P<0.01); The Sham operated AQP4, GFAP expression of grey value, respectively 202.08±5.80, 173.73±4.40, was higher than the control model group (t=46.07, P<0.01,t=31.07,P<0.01); observed under electron microscope, the tight junction of the endothelial cells were integral, the tissue surrounding BBB was in good condition. In the middle group and the treatment group, cerebrovascular endothelial cells connected clearance, even the visible connection integrity of the matrix and basement membrane of the endothelial cells lost.Conclusions rhG-CSF has a protective effect to cerebral ischemia reperfusion injury, the possible mechanism is that it can reduce the expression of GFAP, AQP4 by inhibiting excessive activation, relief brain edema, then protect the integrity of the BBB.
