CAJ | 학술논문

目的对比正常与梗死后大鼠来源的骨髓单个核细胞(Nor-BMM NCs；Isch-BMMNCs)移植治疗大脑中动脉栓塞(MCAO)大鼠的疗效及对血管新生效应的影响.方法线栓法制作大鼠MCAO模型；梯度离心法分别提取Nor-BMMNCs及Isch-BMMNCs;用酶联免疫吸附法(ELISA)检测常氧及低氧条件下体外培养BMMNCs的上清液中VEGF、bFGF、Ang-1的表达水平；MCAOld后行股静脉1 × 107BMMNCs或PBS输注并行mNSS；细胞移植后14 d行TTC染色及免疫荧光检测.结果低氧条件下培养1 d后,Isch-BMMNCs组分泌促血管因子的量[VEGF:(114.2±10.6) pg/ml,bFGF:(133.6 ± 19.7) pg/ml,Ang-1:(131.3±21.9) pg/ml]明显高于Nor-BMMNCs组[VEGF:(96.0±12.6) pg/ml,bFGF:(107.1±19.6) pg/ml,Ang-1:(103.0±17.0) pg/ml],差异有统计学意义(P＜0.05)；mNSS评分显示,在细胞移植后10d,Isch-BMMNCs组神经缺损症状明显改善,与Nor-BMMNCs组及PBS组比较,差异有统计学意义(P＜0.05)；细胞移植14 d后,Isch-BMMNCs组微血管数目及脑梗死体积[(372.5±40.1)支/mm2,(18.7±4.5)％]与Nor-BMMNCs组[(314.0 ±40.1)支/mm2,(26.5±7.2)％]及PBS组[(222.3 ±56.7)个/mm2,(34.5±6.3)％]比较,差异有统计学意义(P＜0.05).结论 Isch-BMMNCs治疗缺血性脑卒中的价值明显高于Nor-BMMNCs,可能与其分泌促血管因子能力增强有关.
목적 대비정상여경사후대서래원적골수단개핵세포(Nor-BMM NCs；Isch-BMMNCs)이식치료대뇌중동맥전새(MCAO)대서적료효급대혈관신생효응적영향.방법 선전법제작대서MCAO모형；제도리심법분별제취Nor-BMMNCs급Isch-BMMNCs;용매련면역흡부법(ELISA)검측상양급저양조건하체외배양BMMNCs적상청액중VEGF、bFGF、Ang-1적표체수평；MCAOld후행고정맥1 × 107BMMNCs혹PBS수주병행mNSS；세포이식후14 d행TTC염색급면역형광검측.결과 저양조건하배양1 d후,Isch-BMMNCs조분비촉혈관인자적량[VEGF:(114.2±10.6) pg/ml,bFGF:(133.6 ± 19.7) pg/ml,Ang-1:(131.3±21.9) pg/ml]명현고우Nor-BMMNCs조[VEGF:(96.0±12.6) pg/ml,bFGF:(107.1±19.6) pg/ml,Ang-1:(103.0±17.0) pg/ml],차이유통계학의의(P＜0.05)；mNSS평분현시,재세포이식후10d,Isch-BMMNCs조신경결손증상명현개선,여Nor-BMMNCs조급PBS조비교,차이유통계학의의(P＜0.05)；세포이식14 d후,Isch-BMMNCs조미혈관수목급뇌경사체적[(372.5±40.1)지/mm2,(18.7±4.5)％]여Nor-BMMNCs조[(314.0 ±40.1)지/mm2,(26.5±7.2)％]급PBS조[(222.3 ±56.7)개/mm2,(34.5±6.3)％]비교,차이유통계학의의(P＜0.05).결론 Isch-BMMNCs치료결혈성뇌졸중적개치명현고우Nor-BMMNCs,가능여기분비촉혈관인자능력증강유관.
Objective To compare the therapeutic effect and angiogenic capability of bone marrow mononuclear cells derived from normal rats and ischemic rats for stroke.Methods The MCAO model was established with suture emboli method ; bone marrow mononuclear cells (BMMNCs) were isolated with density gradient centrifugation method.The level of vascular endothelial growth factor(VEGF),basic fibroblast growth factor(bFGF),angiopoietin-1 (Aug-1) were tested by ELISA under normoxic and hypoxic conditions.The rats were injected with PBS or 1 × l07 BMMNCs via femoral vein at day 1 after stroke.hnmunofluorescence assay was performed at day 14 after cell transplantation.Results When compared with Nor-BMMNCs (VEGF:(96.0 ± 12.6) pg/ml,bFGF:(107.1 ± 19.6)pg/ml,Ang-1:(103.0 ± 17.0)pg/ml),hypoxic stimulation significantly increased the secretion (VEGF:(114.2 ± 10.6)pg/ml,bFGF:(133.6 ± 19.7)pg/mL,Ang-I:(131.3 ± 21.9) pg/ml)at day 1 after cell culture(P＜ 0.05).The neurological deficits in rats of Isch-BMMNCs group were obviously improved compared with those in rats of Nor-BMMNCs group and PBS group at day 10 after transplantation (P ＜ 0.05).In addition,IschBMMNCs also significantly increased the number of microvessels and decreased the inffract volume ((372.5 ±40.1)/mm2,(18.7 ±.4.5) ％) when compared with Nor-BMMNCs group ((314.0 ± 40.1)/mm2,(26.5 ±7.2) ％) and PBS group ((222.3 ± 56.7)/mm2,(34.5 ± 6.3) ％) at day 14 in vivo (P ＜ 0.05).Conclusion Isch-BMMNCs exert more neuroprotective effects than Nor-BMMNCs,which may relate with the difference in the secretion capability of angiogenic factors of this two original cells.