皮肤源性前体细胞经不同方式移植后在大鼠损伤脊髓中存活情况及对功能恢复的影响

Survival of skin-derived precursors in rat's injured spinal cord and its effect on functional recovery after transplantation in different ways

目的:观察皮肤源性前体细胞(SKPs)经不同方式移植在大鼠的损伤脊髓中的存活情况及对功能恢复的作用。方法:分离雄性大鼠SKPs并检测其迁移能力。Allen法制备大鼠脊髓损伤模型后,将DAPI标记的SKPs经局部注射(A组)、尾静脉(B组)、脑脊液(C组)移植到雌性大鼠体内。伤后第3天和第7天取材WesternBlot检测损伤局部趋化因子的表达,荧光显微镜观察和实时定量PCR法检测脊髓中SKPs含量。BBB法评价大鼠功能恢复情况。结果:SKPs具有很强的迁移能力。损伤局部趋化因子的表达上调,移植后第7天B组和C组脊髓SKPs数量较第3天时增加,但均低于A组;而伤后第7天B组和C组大鼠的功能恢复好于A组。结论:经静脉和脑脊液移植的SKPs可在损伤脊髓局部上调表达趋化因子的作用下迁移到损伤局部并存活,功能恢复优于局部注射法。

Objective： To observe the survival characterization of skin-derived precursors （SKPs） in rat＇s injured spinal cord and its effect on functional recovery after transplantation in different ways. Method： SKPs were isolated from male rats and its migration ability was assessed in vitro. After spinal cord injury model was introduced by Allen＇s method, DAPI-labeled SKPs were transplanted by local injection, tail vein injection and via cerebral spinal fluid to re.male rats. Three and seven days after injury, Western Blot was used to examine the expression of chemotactic factors in injured sites, and observation under fluorescent microscope and with real-time PCR were employed to examine the amount of SKPs in injured spinal cord. BBB method was used to assess the functional recovery of rats. Result： SKPs showed potent migration ability. The expression of chemotactic factors was up-regulated in injured sites. At the 7th d after injury, the amount of SKPs in injured spinal cord of group B and C rats increased when cmnpared with the 3th d, which was both lower than that in group A. However, the functional recoveries in group B and C were better than that in group A. Conclusion： SKPs transplanted via vein and cerebral spinal fluid could migrate to injured sites under the effect of up-regulated chemotactic factors and survive, with better functional recovery than local injection.