中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2015年
1期
22-27
,共6页
朱军%李茂琴%方向韶%黄子通
硃軍%李茂琴%方嚮韶%黃子通
주군%리무금%방향소%황자통
心搏骤停%心肺复苏%干细胞动员%大鼠%粒细胞集落刺激因子%神经功能缺损评分
心搏驟停%心肺複囌%榦細胞動員%大鼠%粒細胞集落刺激因子%神經功能缺損評分
심박취정%심폐복소%간세포동원%대서%립세포집락자격인자%신경공능결손평분
Cardiac arrest%Cardiopulmonary resuscitation%Stem cell mobilization%Rats%Granulocyte colony-stimulating factor%Neurological deficit score
目的 观察粒细胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)与AMD3100动员自身骨髓干细胞治疗复苏后脑缺血损伤的效果,并探讨其作用机制.方法 在中山大学心肺脑复苏研究所建立窒息法心肺复苏大鼠动物模型.56只SD大鼠随机(随机数字法)分4组:G-CSF单独动员组、G-CSF+ AMD3100联合动员组、单纯复苏组和假手术组.通过NDS评分、纸带移除实验、头颅MR扫描影像分析等方法评价干细胞自体动员对复苏后神经功能的影响;采用ELISA法检测脑组织中血管内皮细胞生长因子(VEGF)含量、TUNEL法检测脑组织中神经细胞凋亡、免疫荧光法检测脑组织毛细血管密度变化.结果 复苏后3d时,G-CSF+ AMD3100组NDS评分(61.4±10.7)显著高于单纯复苏组(49.9±10.4) (P＜0.05),纸带移除时间为(85.5±28.9)s,显著短于单纯复苏组(148.1±23.8)s与G-CSF组(118.5 ±30.4)s (P＜0.05);脑MRI显示的脑损伤严重程度两个干细胞动员组低于单纯复苏组;G-CSF+ AMD3100组的神经细胞凋亡率(0.23±0.06)显著低于G-CSF组(0.34 ±0.08) (P＜0.05),而两者均显著低于单纯复苏组(0.44±0.09)(P＜0.05).在复苏后3d与6d,G-CSF+ AMD3100组脑组织中VEGF质量浓度(pg/mL)分别为(106.2±23.3)与(79.9±18.4),G-CSF组脑组织VEGF质量浓度(pg/mL)分别为(50.6±13.7)与(73.9±16.6),均显著高于单纯复苏组(23.1±10.2)与(36.2±12.8)(P＜0.05).G-CSF+ AMD3100组在复苏后3d时的脑毛细血管密度(351.8 ±67.9)个/高倍视野,显著高于G-CSF组(301.4 ±77.3)个/高倍视野与单纯复苏组(250.4 ±48.0)个/高倍视野(P＜0.05).在复苏后6d时,G-CSF组的脑毛细血管密度较3d时明显升高,为(348.4 ±76.7)个/高倍视野(P＜0.05),G-CSF+ AMD3100组为(344.1±65.7)个/高倍视野,与3d时比较无明显变化.结论 干细胞动员显著改善了复苏后大鼠神经功能状况,联合动员后神经功能恢复更加显著且恢复时间早于单独动员.干细胞动员对脑损伤的修复作用机制可能与抑制神经细胞凋亡、促进VEGF分泌及损伤区新生血管生成有关.
目的 觀察粒細胞集落刺激因子(granulocyte colony-stimulating factor,G-CSF)與AMD3100動員自身骨髓榦細胞治療複囌後腦缺血損傷的效果,併探討其作用機製.方法 在中山大學心肺腦複囌研究所建立窒息法心肺複囌大鼠動物模型.56隻SD大鼠隨機(隨機數字法)分4組:G-CSF單獨動員組、G-CSF+ AMD3100聯閤動員組、單純複囌組和假手術組.通過NDS評分、紙帶移除實驗、頭顱MR掃描影像分析等方法評價榦細胞自體動員對複囌後神經功能的影響;採用ELISA法檢測腦組織中血管內皮細胞生長因子(VEGF)含量、TUNEL法檢測腦組織中神經細胞凋亡、免疫熒光法檢測腦組織毛細血管密度變化.結果 複囌後3d時,G-CSF+ AMD3100組NDS評分(61.4±10.7)顯著高于單純複囌組(49.9±10.4) (P＜0.05),紙帶移除時間為(85.5±28.9)s,顯著短于單純複囌組(148.1±23.8)s與G-CSF組(118.5 ±30.4)s (P＜0.05);腦MRI顯示的腦損傷嚴重程度兩箇榦細胞動員組低于單純複囌組;G-CSF+ AMD3100組的神經細胞凋亡率(0.23±0.06)顯著低于G-CSF組(0.34 ±0.08) (P＜0.05),而兩者均顯著低于單純複囌組(0.44±0.09)(P＜0.05).在複囌後3d與6d,G-CSF+ AMD3100組腦組織中VEGF質量濃度(pg/mL)分彆為(106.2±23.3)與(79.9±18.4),G-CSF組腦組織VEGF質量濃度(pg/mL)分彆為(50.6±13.7)與(73.9±16.6),均顯著高于單純複囌組(23.1±10.2)與(36.2±12.8)(P＜0.05).G-CSF+ AMD3100組在複囌後3d時的腦毛細血管密度(351.8 ±67.9)箇/高倍視野,顯著高于G-CSF組(301.4 ±77.3)箇/高倍視野與單純複囌組(250.4 ±48.0)箇/高倍視野(P＜0.05).在複囌後6d時,G-CSF組的腦毛細血管密度較3d時明顯升高,為(348.4 ±76.7)箇/高倍視野(P＜0.05),G-CSF+ AMD3100組為(344.1±65.7)箇/高倍視野,與3d時比較無明顯變化.結論 榦細胞動員顯著改善瞭複囌後大鼠神經功能狀況,聯閤動員後神經功能恢複更加顯著且恢複時間早于單獨動員.榦細胞動員對腦損傷的脩複作用機製可能與抑製神經細胞凋亡、促進VEGF分泌及損傷區新生血管生成有關.
목적 관찰립세포집락자격인자(granulocyte colony-stimulating factor,G-CSF)여AMD3100동원자신골수간세포치료복소후뇌결혈손상적효과,병탐토기작용궤제.방법 재중산대학심폐뇌복소연구소건립질식법심폐복소대서동물모형.56지SD대서수궤(수궤수자법)분4조:G-CSF단독동원조、G-CSF+ AMD3100연합동원조、단순복소조화가수술조.통과NDS평분、지대이제실험、두로MR소묘영상분석등방법평개간세포자체동원대복소후신경공능적영향;채용ELISA법검측뇌조직중혈관내피세포생장인자(VEGF)함량、TUNEL법검측뇌조직중신경세포조망、면역형광법검측뇌조직모세혈관밀도변화.결과 복소후3d시,G-CSF+ AMD3100조NDS평분(61.4±10.7)현저고우단순복소조(49.9±10.4) (P＜0.05),지대이제시간위(85.5±28.9)s,현저단우단순복소조(148.1±23.8)s여G-CSF조(118.5 ±30.4)s (P＜0.05);뇌MRI현시적뇌손상엄중정도량개간세포동원조저우단순복소조;G-CSF+ AMD3100조적신경세포조망솔(0.23±0.06)현저저우G-CSF조(0.34 ±0.08) (P＜0.05),이량자균현저저우단순복소조(0.44±0.09)(P＜0.05).재복소후3d여6d,G-CSF+ AMD3100조뇌조직중VEGF질량농도(pg/mL)분별위(106.2±23.3)여(79.9±18.4),G-CSF조뇌조직VEGF질량농도(pg/mL)분별위(50.6±13.7)여(73.9±16.6),균현저고우단순복소조(23.1±10.2)여(36.2±12.8)(P＜0.05).G-CSF+ AMD3100조재복소후3d시적뇌모세혈관밀도(351.8 ±67.9)개/고배시야,현저고우G-CSF조(301.4 ±77.3)개/고배시야여단순복소조(250.4 ±48.0)개/고배시야(P＜0.05).재복소후6d시,G-CSF조적뇌모세혈관밀도교3d시명현승고,위(348.4 ±76.7)개/고배시야(P＜0.05),G-CSF+ AMD3100조위(344.1±65.7)개/고배시야,여3d시비교무명현변화.결론 간세포동원현저개선료복소후대서신경공능상황,연합동원후신경공능회복경가현저차회복시간조우단독동원.간세포동원대뇌손상적수복작용궤제가능여억제신경세포조망、촉진VEGF분비급손상구신생혈관생성유관.
Objective To explore the therapeutic potential and mechanism of stem cells mobilized by granulocyte colony-stimulating factor (G-CSF) and AMD3100 to repair global cerebral ischemia injuries in a rat model of cardiac arrest (CA) and cardiopulmonary resuscitation (CPR).Methods Cardiac arrest was induced by asphyxia.Fifty-six SD rats were randomly assigned into four groups:G-CSF group,G-CSF + AMD3100 group,CPR control group and sham operated group.The animals were sacrificed at 3d and 6d after CPR respectively.The neurological status and morphological changes of damaged cerebrum,the apoptosis of nerve cells and vascular endothelial growth factor (VEGF) expressed in brain tissue and capillary density in hippocampus and temporal lobe cortex were measured and analyzed by means of neurological deficit score (NDS),adhesive tape removal test (TRT),ELISA,MRI and immunofluorescence.Results NDS in G-CSF + AMD3100 group (61.4 ± 10.7) was significantly higher than that in CPR control group (49.9 ± 10.4) at 3 d after CPR (P ＜0.05).And less time consumption for TRT found in G-CSF + AMD3100 group (85.5 ±28.9) s rather than was in CPR control group (148.1 ± 23.8) s and G-CSF group (118.5 ± 30.4) s (P ＜ 0.05).The severity of cerebral injury assessed by MRI was significantly milder at both 3 d and 6 d in the two stem cell mobilization groups.The apoptosis rate of nerve cells in G-CSF + AMD3100 group (0.23 ± 0.06) was significantly lower than that in G-CSF group (0.34 ±0.08) at 3 d after CPR,and that in both stem cell mobilization groups was lower than that in CPR control group (0.44 ± 0.09) (P ＜ 0.05).At 3 d and 6 d after CPR,the levels of VEGF in brain tissue were (106.2 ±23.3) pg/mL and (79.9 ± 18.4) pg/mL in G-CSF + AMD3100 group,and were (50.6 ± 13.7) pg/mL and (73.9 ± 16.6) pg/mL in G-CSF group,which were both significantly higher than that in CPR control group (23.1 ± 10.2) pg/mL and (36.2 ± 12.8) pg/mL (P ＜0.05).At 3 d after CPR,the cerebral capillary density (351.8 ±67.9) branches in every high power field (A/HPF) was significantly higher in G-CSF + AMD3100 group than that (301.4 ± 77.3) A/HPF in G-CSF group and (250.4 ± 48.0) A/HPF in CPR control group (P ＜ 0.05).The cerebral capillary density in G-CSF group elevated to (348.4 ±76.7) A/HPF at 6 d after CPR which was significantly higher than that at 3 d (P ＜0.05),and there was no difference between that at 3 d and 6 d in G-CSF + AMD3100 group.Conclusions The mobilization stem cells improve the impaired neurological function.The increased expression of VEGF in brain tissue,the neo-vascularization promoted by the mobilized stem cells and the inhibition of nerve cell apoptosis may be associated with the protective effects of the stem cell mobilization.
