中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Trauma
2015年
11期
1014-1019
,共6页
颅脑损伤%间质干细胞%移植%治疗
顱腦損傷%間質榦細胞%移植%治療
로뇌손상%간질간세포%이식%치료
Craniocerebral trauma%Mesenchymal stem cells%Transplantation%Therapy
目的 探讨大鼠创伤性脑损伤(TBI)后,采用骨髓间充质干细胞(BMSCs)诱导产生血管内皮祖细胞(EPCs)移植治疗大鼠TBI的作用,以及EPCs移植相对于BMSCs移植的优越性.方法 选取3个月龄、体重约150 g雄性SD大鼠获取BMSCs,诱导培养EPCs,通过CD133、CD34及FLK-1的表达对EPCs进行鉴定.选取成年雌性SD大鼠120只,按随机概率表法分为EPCs组、BMSCs组和成纤维细胞(3T3)组,每组40只.液压冲击TBI装置制备大鼠中、重度TBI模型.伤后1d各组分别经尾静脉移植BrdU标记的EPCs、BMSCs或3T3细胞.移植后2,7,14,28 d采用改良神经严重度评分(mNSS)评估神经功能恢复程度,取损伤脑组织行免疫组化检测各组CD34、脑源性生长因子(BDNF)阳性细胞数,评估神经和血管再生情况.利用荧光免疫原位杂交(FISH)技术检测Sry基因阳性细胞数. 结果 CD133、CD34及FLK-1阳性率分别为52％、33％和38％,表明BMSCs是具有EPCs表型的细胞.移植7,14,28 d EPCs组mNSS为(10.2±1.5)分、(8.7±1.0)分、(4.9±1.0)分,BMSCs组为(10.8±1.7)分、(10.1±1.7)分、(7.2±1.3)分,均明显低于3T3组的(12.4±1.5)分、(11.7±1.8)分、(10.3±1.5)分(P＜0.05).BrdU标记法结果显示,移植14,28 d EPCs组CD34、BDNF阳性细胞表达数EPCs组均明显高于BMSCs组(P＜0.05),3T3组均未见CD34、BDNF阳性细胞表达.移植细胞示踪的Sry探针法比BrdU标记法显示的荧光阳性细胞数更多. 结论 BMSCs和EPCs经尾静脉移植后均可归巢至大鼠TBI处,EPCs治疗效果更好.
目的 探討大鼠創傷性腦損傷(TBI)後,採用骨髓間充質榦細胞(BMSCs)誘導產生血管內皮祖細胞(EPCs)移植治療大鼠TBI的作用,以及EPCs移植相對于BMSCs移植的優越性.方法 選取3箇月齡、體重約150 g雄性SD大鼠穫取BMSCs,誘導培養EPCs,通過CD133、CD34及FLK-1的錶達對EPCs進行鑒定.選取成年雌性SD大鼠120隻,按隨機概率錶法分為EPCs組、BMSCs組和成纖維細胞(3T3)組,每組40隻.液壓遲擊TBI裝置製備大鼠中、重度TBI模型.傷後1d各組分彆經尾靜脈移植BrdU標記的EPCs、BMSCs或3T3細胞.移植後2,7,14,28 d採用改良神經嚴重度評分(mNSS)評估神經功能恢複程度,取損傷腦組織行免疫組化檢測各組CD34、腦源性生長因子(BDNF)暘性細胞數,評估神經和血管再生情況.利用熒光免疫原位雜交(FISH)技術檢測Sry基因暘性細胞數. 結果 CD133、CD34及FLK-1暘性率分彆為52％、33％和38％,錶明BMSCs是具有EPCs錶型的細胞.移植7,14,28 d EPCs組mNSS為(10.2±1.5)分、(8.7±1.0)分、(4.9±1.0)分,BMSCs組為(10.8±1.7)分、(10.1±1.7)分、(7.2±1.3)分,均明顯低于3T3組的(12.4±1.5)分、(11.7±1.8)分、(10.3±1.5)分(P＜0.05).BrdU標記法結果顯示,移植14,28 d EPCs組CD34、BDNF暘性細胞錶達數EPCs組均明顯高于BMSCs組(P＜0.05),3T3組均未見CD34、BDNF暘性細胞錶達.移植細胞示蹤的Sry探針法比BrdU標記法顯示的熒光暘性細胞數更多. 結論 BMSCs和EPCs經尾靜脈移植後均可歸巢至大鼠TBI處,EPCs治療效果更好.
목적 탐토대서창상성뇌손상(TBI)후,채용골수간충질간세포(BMSCs)유도산생혈관내피조세포(EPCs)이식치료대서TBI적작용,이급EPCs이식상대우BMSCs이식적우월성.방법 선취3개월령、체중약150 g웅성SD대서획취BMSCs,유도배양EPCs,통과CD133、CD34급FLK-1적표체대EPCs진행감정.선취성년자성SD대서120지,안수궤개솔표법분위EPCs조、BMSCs조화성섬유세포(3T3)조,매조40지.액압충격TBI장치제비대서중、중도TBI모형.상후1d각조분별경미정맥이식BrdU표기적EPCs、BMSCs혹3T3세포.이식후2,7,14,28 d채용개량신경엄중도평분(mNSS)평고신경공능회복정도,취손상뇌조직행면역조화검측각조CD34、뇌원성생장인자(BDNF)양성세포수,평고신경화혈관재생정황.이용형광면역원위잡교(FISH)기술검측Sry기인양성세포수. 결과 CD133、CD34급FLK-1양성솔분별위52％、33％화38％,표명BMSCs시구유EPCs표형적세포.이식7,14,28 d EPCs조mNSS위(10.2±1.5)분、(8.7±1.0)분、(4.9±1.0)분,BMSCs조위(10.8±1.7)분、(10.1±1.7)분、(7.2±1.3)분,균명현저우3T3조적(12.4±1.5)분、(11.7±1.8)분、(10.3±1.5)분(P＜0.05).BrdU표기법결과현시,이식14,28 d EPCs조CD34、BDNF양성세포표체수EPCs조균명현고우BMSCs조(P＜0.05),3T3조균미견CD34、BDNF양성세포표체.이식세포시종적Sry탐침법비BrdU표기법현시적형광양성세포수경다. 결론 BMSCs화EPCs경미정맥이식후균가귀소지대서TBI처,EPCs치료효과경호.
Objective To observe if endothelial progenitor cells (EPCs) and bone marrow mesenchymal stem cells (BMSCs) could home to the injured region and study the effect of BMSCs-derived EPCs on traumatic brain injury (TBI) in rats.Methods BMSCs were isolated from 3-month-old SD male rats weighing 150 g,and induced to EPCs.EPCs were identified by surface markers CD133,CD34 and FLK-1.A total of 120 female adult SD rats were divided into 3 groups (n =40 each) according to the random probability table method:EPCs group,BMSCs group and 3T3 cells group.The model of moderate to severe TBI was induced by the fluid percussion device.All the cells (i.e.EPCs,BMSCs and 3T3 cells) were injected with BrdU before transplantation to the tail vein of rats.On days 2,7,14 and 28 after transplantation,rat neurological function was evaluated using the modified neurological severity score (mNSS),and injured brain tissue was harvested to detect CD34 and brain-derived neurotrophic factor (BDNF) positive cell density using the immunohistochemistry method.Sry-positive cells were evaluated using the fluorescence in situ hybridization (FISH).Results The positive rate of CD133,CD34 and Flk-1 was 52％,33％ and 38％,indicating BMSCs differentiation towards an EPCs phenotype.On days 7,14 and 28 after transplantation,the mNSS in EPCs group [(10.2 ± 1.5),(8.7 ± 1.0) and (4.9 ± 1.0) points] and in BMSCs group [(10.8 ± 1.7),(10.1 ± 1.7),and (7.2 ± 1.3) points] were lower than that in 3T3 cells group [(12.4 ± 1.5),(1 1.7 ± 1.8),(10.3 ± 1.5) points] (P ＜ 0.05).On 14 and 28 days after transplantation,BrdU-positive CD34 and BDNF in EPCs group were significantly more than those in BMSCs group (P ＜ 0.05).Instead,there were no positive cells in 3T3 cells group.The method of Sry probe to trace the transplanted cells could detect more positive cells compared to the BrdU labeling method.Conclusion Both BMSCs and EPCs can home to the injured region,but EPCs have much better therapeutic effect.
