大鼠骨髓间充质干细胞条件培养基对2型糖尿病大鼠脑缺血／再灌注损伤后血管新生的影响

Effects of bone mesenchymal stem cell conditioned medium on angiogenesis in type 2 diabetic rats after ischemia/reperfusion

目的：探讨大鼠骨髓间充质干细胞条件培养基（ BMSCs-CM）对2型糖尿病大鼠脑缺血/再灌注损伤后缺血半暗带区血管新生的影响。方法取雄性Wistar大鼠股骨骨髓作为供体来源制备BMSCs-CM。链脲佐菌素-烟酰胺诱导2型糖尿病大鼠模型，成功后用线栓法制备大脑中动脉缺血/再灌注损伤模型，缺血2h后血管再通。随机分为生理盐水对照组（ NS组）、单纯培养基对照组（ DMEM组）、骨髓间充质干细胞条件培养基组（CM组）。术后2、24、48 h，CM组大鼠经尾静脉给予BMSCs-CM（10 ml/kg），DMEM组给予DMEM（10 ml/kg）， NS组给予生理盐水（10 ml/kg）。术后14天，分别比较3组大鼠脑缺血半暗带区血管性血友病因子（ vWF）和血管生成素-1（ Ang1）的变化情况。结果术后14天，CM组与其他2组相比，脑缺血半暗带区vWF阳性血管密度及阳性血管周长均明显增加，差异有统计学意义（P＜0．05），而NS组、DMEM组相比，差异无统计学意义（P＞0．05）；CM组与其他2组相比，大鼠脑缺血半暗带区Ang1表达量显著增加，差异有统计学意义（P＜0．01），而NS组、DMEM组相比，差异无统计学意义（P＞0．05）。结论大鼠BMSCs-CM可促进2型糖尿病大鼠脑缺血/再灌注损伤后缺血半暗带区血管新生。

Objective To investigate the effects of bone mesenchymal stem cell conditioned medium （ BMSC-CM） on the angiogenesis of type 2 diabetic （T2DM） rats after ischemia/reperfusion.Methods BMSC-CM was prepared using the bone marrow from the fermur of male wistar rats .A model of type 2 diabetic rats was established by injection of streptozotocin-nicotinamide.Then the obtained T2DM rats were subjected to middle cerebral artery occlusion （MCAo） using a nylon monofilament suture followed by reperfusion 2 hours later.After surgery, the T2DM ischemic rats were randomly assigned to three groups according to intravenous injection of 10 ml/kg normal saline （ NS） , 10 ml/kg DMEM culture medium or 10 ml/kg BMSC-CM through their tail vein 2, 24 and 48 hours after surgery.On day 14, the changes of von Willebrand factor （vWF） and angiopoietin 1 （Ang1） in the ischemic penumbra were detected by immunostaining . Results Compared to the NS and DMEM groups , the CM group presented significant increases in vWF positive vascular density and vascular perimeter （P〈0.05）.However, there was no significant difference between the NS and DMEM groups （P〉0.05）.Compared to the NS and DMEM groups, remarkably increased amounts of Ang1 were detected in the CM group （P〈0.01）.However, there was no significant difference between the NS and DMEM groups as to Ang 1 expression （P〉0.05）.Conclusion Bone mesenchymal stem cell conditioned medium can stimulate the angiogenesis within the ischemic penumbra of T2DM rats after ischemia/reperfusion.