摘要
目的:观察骨髓间充质干细胞(BMSCs)移植联用褪黑素(MT)对大鼠脊髓损伤修复的影响,并对其作用机制进行探讨。方法:首先采用密度梯度离心法分离培养大鼠BMSCs,以Brdu标记细胞核。然后将48只SD大鼠用改良Allen法制备T10脊髓损伤模型,并随机平均分为MT+BMSCs组、BMSCs组、MT组、模型对照组,另取12只作为空白对照组。分别于造模后1d、3d、7d、14d、21d行神经功能BBB评分及斜板试验,免疫组化检测带标记的BMSCs迁移情况,显微镜下观察移植后脊髓形态结构变化。结果:术后神经功能BBB评分及斜板试验结果MT+BMSCs组优于BMSCs组(P<0.05),BMSCs组优于MT及模型对照组(P<0.05);MT+BMSCs组及BMSCs组损伤阶段脊髓内可见Brdu标记阳性BMSCs存活,且阳性数目前者多于后者,灰质分布较白质多,以注射部位向周围损伤组织迁移;同时显微镜下可观察到MT+BMSCs组脊髓损伤区无明显空腔,空泡小而少,并可见较多神经纤维及细胞。结论:BMSCs联合MT移植促进大鼠损伤脊髓结构和功能恢复的效果明显优于单纯细胞移植组,两者联用具有协同效应。
Objective To observe the influence of BMSCs transplantation combined with melatonin on repair of spinal cord injury in rats, and to explore its possible mechanism. Methods BMSCs were isolated and cultured with density gradient centrifugation from rats and the nucleus of BMSCs were labelled with Brud. The T10 spinal cord injury model was prepared with modified Allen method in 48 rats, which were randomly divided into MT ＋ BMSCs group, BMSCs group, MT group and model group ,and another 12 healthy rats were selected as control group. The BBB scores of neurologic function and inclined plane test were detected at 1 d,3 d,7 d,14 d,21 d after operation,the migration of labelled BMSCs were determined with immunohistochemical and the change of morphology after transplantation was observed under microscope. Results The re- suits of BBB scores and inclined plane test showed that MT ＋ BMSCs group were significantly better than those of BMSCs group （ P 〈 0.05 ） , BMSCs group were significantly better than those of MT group and model group （ P 〈 0.05 ）. The survival postively labelled BMSCs were observed in injuried spinal cord in MT ＋ BMSCs group and BMSCs group, and the number of survival cells in former group was than the latter, in gray matter was more than that of white matter. The survival BMSCs was migrated from injected location to injuried area. Nerve fibers and cells were observed under microscope in spinal cord inju- ried area of MT ＋ BMSCs group, while no obvious empty cavity was observed. Conclusion BMSCs transplantation combined MT could promote structure and functional recovery of spinal cord injury with a synergistic effect, and the effect was better than single transplantation.
出处
《郧阳医学院学报》
2010年第1期5-8,F0003,共5页
Journal of Yunyang Medical College
关键词
脊髓损伤
骨髓间充质干细胞
褪黑素
移植
Spinal cord injury
Bone mesenchymal stem ceU
Melatonin
Transplantation
