国际脑血管病杂志
國際腦血管病雜誌
국제뇌혈관병잡지
INTERNATIONAL JOURNAL OF CEREBROVASCULAR DISEASES
2012年
4期
257-262
,共6页
孔朝红%刘煜敏%朱江%周绍霞
孔朝紅%劉煜敏%硃江%週紹霞
공조홍%류욱민%주강%주소하
脑缺血%干细胞%内皮,血管%细胞移植%新生血管化,生理性%血管内皮生长因子%细胞凋亡%疾病模型,动物%大鼠
腦缺血%榦細胞%內皮,血管%細胞移植%新生血管化,生理性%血管內皮生長因子%細胞凋亡%疾病模型,動物%大鼠
뇌결혈%간세포%내피,혈관%세포이식%신생혈관화,생이성%혈관내피생장인자%세포조망%질병모형,동물%대서
Brain ischemia%Stem Cells%Endothelium,Vascular%Cell Transplantation%Neovascularization,Physiologic%Vascular Endothelial Growth Factors%Apoptosis%Disease Models,Animal%Rats
目的 探讨目体骨髓米源内皮祖细肥(endothehalprogenitor cell,EPC)移植对脑缺血大鼠神经功能转归的影响及其可能机制.方法 体外分离培养自体骨髓来源EPC并用5-溴脱氧尿嘧啶核苷(5 -bromodeoxyuridine,BrdU)标记.线栓法制作大鼠大脑中动脉闭塞(middle cerebral arteryocclusion,MCAO)模型.EPC组大鼠经颈外静脉移植自体EPC[ 106/ml·kg)],对照组注射磷酸盐缓冲液(1 ml/kg),假手术组不进行任何处理(n=15).改良神经功能缺损严重程度评分(modifiedneurological severity score,mNSS)观察大鼠神经功能变化情况.BrdU免疫组化染色评价EPC增殖和分化.三维共聚焦图像分析检测脑缺血区血管结构和密度.TUNEL染色检测缺血脑组织凋亡细胞.酶联免疫吸附法检测血浆血管内皮生长因子(vascular endothelial growth factor,VEGF)浓度.结果 EPC组mNSS评分显著低于对照组[第8天时；(6.43±0.69)分对(8.86±0.95)分；q=2.673,P=0.035；第14天时:(4.55±0.89)分对(6.73±1.06)分；q=5.360,P=0.035].EPC组BrdU阳性细胞数量显著多于对照组[(42.2±5.76)对(25.67±5.49)；q=4.020,P=0.030].EPC组毛细血管直径显著小于对照组[(4.51±0.21)μm对(6.34±0.24) μm;q=3.980,P =0.003]；血管密度[(212.64±8.02)/0.002 mm3对(153.60±7.21 )/0.002 mn3；q =9.670,P=0.001]和微血管总表面积[(92 013±5 132)μm3/0.002 mm3对(71 366±4 538) μm2/0.002 mm3；q=4.180,P=0.014]显著高于和大于对照组；EPC组凋亡细胞数量显著少于对照组[(36.26±6.91)对(78.34±7.21)；t=-4.834,P=0.003]；EPC组血浆VEGF浓度显著高于对照组[(54.91±5.71)pg/ml对(13.81±4.25)pg/ml;q=12.300,P=0.002].结论 自体EPC移植对大鼠缺血脑组织具有保护作用,可能与VEGF相关联的血管再生和神经保护有关,其在治疗缺血性脑血管病中具有重要的应用前景.
目的 探討目體骨髓米源內皮祖細肥(endothehalprogenitor cell,EPC)移植對腦缺血大鼠神經功能轉歸的影響及其可能機製.方法 體外分離培養自體骨髓來源EPC併用5-溴脫氧尿嘧啶覈苷(5 -bromodeoxyuridine,BrdU)標記.線栓法製作大鼠大腦中動脈閉塞(middle cerebral arteryocclusion,MCAO)模型.EPC組大鼠經頸外靜脈移植自體EPC[ 106/ml·kg)],對照組註射燐痠鹽緩遲液(1 ml/kg),假手術組不進行任何處理(n=15).改良神經功能缺損嚴重程度評分(modifiedneurological severity score,mNSS)觀察大鼠神經功能變化情況.BrdU免疫組化染色評價EPC增殖和分化.三維共聚焦圖像分析檢測腦缺血區血管結構和密度.TUNEL染色檢測缺血腦組織凋亡細胞.酶聯免疫吸附法檢測血漿血管內皮生長因子(vascular endothelial growth factor,VEGF)濃度.結果 EPC組mNSS評分顯著低于對照組[第8天時；(6.43±0.69)分對(8.86±0.95)分；q=2.673,P=0.035；第14天時:(4.55±0.89)分對(6.73±1.06)分；q=5.360,P=0.035].EPC組BrdU暘性細胞數量顯著多于對照組[(42.2±5.76)對(25.67±5.49)；q=4.020,P=0.030].EPC組毛細血管直徑顯著小于對照組[(4.51±0.21)μm對(6.34±0.24) μm;q=3.980,P =0.003]；血管密度[(212.64±8.02)/0.002 mm3對(153.60±7.21 )/0.002 mn3；q =9.670,P=0.001]和微血管總錶麵積[(92 013±5 132)μm3/0.002 mm3對(71 366±4 538) μm2/0.002 mm3；q=4.180,P=0.014]顯著高于和大于對照組；EPC組凋亡細胞數量顯著少于對照組[(36.26±6.91)對(78.34±7.21)；t=-4.834,P=0.003]；EPC組血漿VEGF濃度顯著高于對照組[(54.91±5.71)pg/ml對(13.81±4.25)pg/ml;q=12.300,P=0.002].結論 自體EPC移植對大鼠缺血腦組織具有保護作用,可能與VEGF相關聯的血管再生和神經保護有關,其在治療缺血性腦血管病中具有重要的應用前景.
목적 탐토목체골수미원내피조세비(endothehalprogenitor cell,EPC)이식대뇌결혈대서신경공능전귀적영향급기가능궤제.방법 체외분리배양자체골수래원EPC병용5-추탈양뇨밀정핵감(5 -bromodeoxyuridine,BrdU)표기.선전법제작대서대뇌중동맥폐새(middle cerebral arteryocclusion,MCAO)모형.EPC조대서경경외정맥이식자체EPC[ 106/ml·kg)],대조조주사린산염완충액(1 ml/kg),가수술조불진행임하처리(n=15).개량신경공능결손엄중정도평분(modifiedneurological severity score,mNSS)관찰대서신경공능변화정황.BrdU면역조화염색평개EPC증식화분화.삼유공취초도상분석검측뇌결혈구혈관결구화밀도.TUNEL염색검측결혈뇌조직조망세포.매련면역흡부법검측혈장혈관내피생장인자(vascular endothelial growth factor,VEGF)농도.결과 EPC조mNSS평분현저저우대조조[제8천시；(6.43±0.69)분대(8.86±0.95)분；q=2.673,P=0.035；제14천시:(4.55±0.89)분대(6.73±1.06)분；q=5.360,P=0.035].EPC조BrdU양성세포수량현저다우대조조[(42.2±5.76)대(25.67±5.49)；q=4.020,P=0.030].EPC조모세혈관직경현저소우대조조[(4.51±0.21)μm대(6.34±0.24) μm;q=3.980,P =0.003]；혈관밀도[(212.64±8.02)/0.002 mm3대(153.60±7.21 )/0.002 mn3；q =9.670,P=0.001]화미혈관총표면적[(92 013±5 132)μm3/0.002 mm3대(71 366±4 538) μm2/0.002 mm3；q=4.180,P=0.014]현저고우화대우대조조；EPC조조망세포수량현저소우대조조[(36.26±6.91)대(78.34±7.21)；t=-4.834,P=0.003]；EPC조혈장VEGF농도현저고우대조조[(54.91±5.71)pg/ml대(13.81±4.25)pg/ml;q=12.300,P=0.002].결론 자체EPC이식대대서결혈뇌조직구유보호작용,가능여VEGF상관련적혈관재생화신경보호유관,기재치료결혈성뇌혈관병중구유중요적응용전경.
Objective To investigate the effect of autologous bone marrow-derived endothelial progenitor cell (EPC) transplantation on neurological outcomes in cerebral ischernia in rats and its poss le mechanisms.Methods Autologous bone marrow-derived EPC was cultured in vitro and it was labeled with 5-bromodeoxyuridine (BrdU).A middle cerebral artery occlusion (MCAO) model was induced by the intraluminal suture method.The rats in a EPC group transplanted autologous EPC (106/ml/kg) via external jugular veins,those in a control group were injected with phosphate buffered saline (1 ml/kg),and those in a sham operation group (n =15)were not treated.The modified neurological severity score (mNSS) was used to observe the neurological changes of the rats.BrdU immunohistochemical staining was used to evaluate EPC proliferation and differentiation.Three-dimensional confocal image analysis was used to detect the vascular structure and density in cerebral ischemic areas.TUNEL staining was used to detect the apoptotio cells in ischernic brain tissue.Enzyme-linked immunosorbent assay was used to detect the concentration of plasma vascular endothelial growth factor VEGF).Results The mNSS in the EPC group was siginficantly lower than that in the control group (at day 8:6.43 ±0.69 vs.8.86 ±0.95,q =2.673,P=0.035; at day 14:4.55 ±0.89 vs.6.73 ± 1.06,q =5.360,P =0.035).The number of BrdU positive cells in the EPC group was significantly higher than that in the control group (42.2±5.76 vs.25.67±5.49,q=4.020,P=0.030).The capiilary diameter in the EPC group was significantly smaller than that in the control group (4.51 ± 0.21 μm vs.6.34 ± 0.24 μm,q =3.980,P =0.003); the density of blood vessels (212.64 ± 8.02/0.002 mm3 vs.153.60 ± 7.21/0.002 mm3; q =9.670,P =0.001 ) and the total surface area of microvessel (92 013 ± 5 132 μm2/0.002 mm3 vs. 71 366 ±4 538 μm2/0.002 mm3; q=4.180,P=0.014) were significantly higher or more than those in the control group.The number of apoptotic cells in the EPC group was significantly less than that in the control group (36.26 ± 6.91 vs.78.34 ± 7.21; t =-4.834,P =0.003).The plasma VEGF concentration in the EPC group was significantly higher than that in the control group (54.91 ± 5.71 pg/ml vs.13.81 ± 4.25 pg/ml,q =12.300,P=0.002).Conclusions Autologous EPC transplantation has a protective effect on ischemic brain tissue in rats.It may be associated with VEGF related angiogenesis and neuroprotection.It has an important application prospect in the treatment of ischemic cerebrovascular disease.
