红花黄素对骨髓间充质干细胞移植修复大鼠脊髓损伤的影响

The Influence of Safflower Yellow in Repairing Spinal Cord Injury with BMSCs Transplantation in Rats

目的:通过研究红花黄素对骨髓间充质干细胞(BMSCs)移植修复大鼠脊髓损伤(SCI)的影响,探讨其可能的作用机制。方法:骨髓细胞贴壁法分离、培养、扩增BMSCs,并进行腺病毒介导的绿色荧光蛋白(GFP)标记。45只SD大鼠行改良Allen法制备SCI模型后,随机分为红花黄素组、单纯BMSCs组和PBS组,红花黄素组于损伤脊髓局部注入GFP-BMSCs,腹腔注射红花黄素40 mg/kg;单纯BMSCs组脊髓注入GFP-BMSCs,PBS组注入同体积0.9%氯化钠溶液。术后1、2、3、4、7 d检测损伤脊髓组织超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量;分别于7、14、21、28 d检测损伤脊髓组织神经元特异性核蛋白(NeuN)和神经生长因子(NGF)表达,BBB评分法进行后肢运动功能评分。结果:红花黄素组BBB评分、脊髓损伤局部SOD活性及NeuN和NGF的表达均明显高于单纯BMSCs组和PBS组(P<0.05),而MDA含量较低(P<0.05)。结论:红花黄素通过抑制脊髓自由基生成、减轻脂质过氧化、上调NeuN和NGF表达而增强骨髓间充质干细胞移植修复大鼠脊髓损伤的效果。

Objective To investigate the influence of safflower yellow(SY) in repairing the spinal cord injury(SCI) by bone marrow mesenchymal stem cells(BMSCs) transplantation in rats and to explore its possible mechanism.Methods BMSCs were isolated,cultured and generated in vitro,then marked with green fluorescin protein mediated with adenovirus vector(Adv-GFP).The SCI model were prepared in 45 SD rats with modified Allen method and then which were randomly divided into SY group,BMSCs group and PBS group.The BMSCs marked with Adv-GFP were injected into spinal locally and SY were intraperitoneally injected with 40 mg/kg in SY group.The GFP marked BMSCs were injected into spinal in BMSCs group,only the same volume of 0.9% sodium chloride solution were given in PBS group.The contents of SOD and MDA in injured spinal cord were determined at 1st,2nd,3rd,4th,7th day postoperation,the expression of neuron specific nuclear protein(NeuN) and nerve growth factor(NGF) were detected at 7th,14th,21st,28th day,the motor neurological function of hind limb were assessed with BBB scores.Results The BBB scores,the local activity of SOD,the expression of NeuN and NGF were all significantly higher,while the content of MDA was significantly lower in SY group than those of BMSCs group and PBS group(allP<0.05).Conclusion Safflower yellow could enhance the repairing effect of BMSCs transplantation in SCI rats by inhibiting the free radicals and adip-peroxidation,upregulating the expression of NeuN and NGF.