正常与脑梗死后大鼠来源的骨髓单个核细胞治疗缺血性脑卒中的效果对比

Comparison of bone marrow mononuclear cells derived from stroke and normal rats for stroke treat-ment

目的对比正常与梗死后大鼠来源的骨髓单个核细胞（Nor—BMMNCs；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；MCAO1d后行股静脉1×10^7BMMNCs或PBS输注并行mNSS；细胞移植后14d行TTC染色及免疫荧光检测。结果低氧条件下培养1d后，Isch—BMMNCs组分泌促血管因子的量[VEGF：（114．24±10．6）pg／ml，bFGF：（133．6±19．7）pg／ml，Ang-1：（131．3±21．9）pg／m1]明显高于Nor—BMMNCs组[VEGF：（96．04±12．6）pg／ml，bFGF：（107．1±19．6）pg／ml，Ang-1：（103．04±17．0）pg／m1]，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14d后，Isch—BMMNCs组微血管数目及脑梗死体积[（372．5±40．1）支／mm^2，（18．7±4．5）％]与Nor—BMMNCs组[（314．0±40．1）支／mm^2，（26．5±7．2）％]及PBS组[（222．3±56．7）个／mm^2，（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。

Objective To compare the therapeutic effect and angiogenic capability of bone marrow mono- nuclear ceils 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 centrif-ugation method. The level of vascular endothelial growth factor（ VEGF）, basic fibroblast growth factor（ bFGF）, an-giopoietin-1 （ Ang-1 ） were tested by ELISA under normoxic and hypoxic conditions. The rats were injected with PBS or 1 × 10^7 BMMNCs via femoral vein at day 1 after stroke. Immunofluorescence 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-1 ： （ 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, Isch-BMMNCs also significantly increased the number of microvessels and decreased the infract volume （（ 372.5 ± 40. 1 ）/mm^2, （ 18.7 ± 4.5 ） % ） when compared with Nor-BMMNCs group （ （ 314.0 ＋ 40. 1 ）/mm^2, （ 26.5 ± 7.2） % ） and PBS group（ （222.3 ± 56.7 ）/mm^2, （ 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 ceils.