骨髓基质干细胞促进大鼠缺血皮瓣血管生成实验研究

Effects of bone mesenchymal stem cell in the formation of angiogenesis in ischemic flap in rats

目的观察骨髓基质干细胞(BMSCs)促进大鼠缺血皮瓣血管生成作用。方法从6只幼年雄性SPF级SD大鼠骨髓中分离获取BMSCs,培养扩增后,取第3代BMSCs。将48只成年SPF级SD大鼠随机分成实验组和对照组,每组24只。在两组大鼠背部皮肤设计超长随意皮瓣,制作皮瓣模型,实验组大鼠注射重悬BMSCs于皮瓣远端,对照组大鼠同法注射等容积的完全培养液,两组大鼠分别于术后7、14 d切取皮瓣远端标本行苏木精-伊红(HE)染色和免疫组织化学染色,检测皮瓣内微血管数及血管内皮生长因子(VEGF)的表达。结果实验组大鼠注射移植术后7、14 d VEGF表达计分数(4.21±1.02、3.88±1.08)分别高于对照组术后7、14 d VEGF表达计分数(2.58±0.97、2.29±1.08),两组比较差异均有统计学意义(t=5.642、5.082,P均<0.05)。实验组术后7、14 d微血管数(72.33±15.60、80.96±12.18)分别高于对照组术后7、14 d微血管数(50.58±15.67、64.00±16.16),两组比较差异均有统计学意义(t=4.820、4.111,P均<0.05)。结论 BMSCs能分化内皮细胞成分,促进大鼠皮瓣血管新生。

Objective To observe the effect of accelerating angiogenesis in ischemic flap with bone marrow-derived mesenchymal stem cells（BMSCs）. Methods BMSCs were isolated from the marrow of Sprague-Dawley（SD）rats and cultured in vitro by adhering method,and the third passage cells were used in the experiment. The model of hyper-proportion flap was obtained on SD rat＇s back,and forty-eight SD rats were randomized equally into control group and experimental group. To contrast with the experimental group and the control group,which was treated by subcutaneously injected with BMSCs,the other group with injecting culture medium of BMSCs. The microvascular density and VEGF expression in flap specimens were measured with HE staining and immunohistochemistry in 7,14 postoperative days respectively. Results The VEGF expression levels of experimental group were 4. 21 ± 1. 02、3. 88 ± 1. 08 in 7,14 postoperative days respectively,and those in control group were 2. 58 ± 0. 97、2. 29 ± 1. 08. In experimental group,the microvascular density of flap specimens were 72. 33 ± 15. 60、80. 96 ± 12. 18 in 7,14 postoperative days＆amp;nbsp;respectively,and those in control group were 50. 58 ± 15. 67、64. 00 ± 16. 16. There were significant difference between experimental group and control group both in microvascular density and VEGF expression levels of flap specimens（P 〈 0. 05）. Conclusions BMSCs can significantly enhance the angiogenesis of ischemic skin flap in rat.