神经干细胞移植联合PEP-1-SOD1对颅脑损伤大鼠神经功能恢复的影响
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常州市卫生局重大科技项目(ZD201212)


Effects of PEP-1-SOD1 combined with neural stem cells transplantation on the neurologic function of rats with traumatic brain injury
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    摘要:

    目的:探讨神经干细胞(neural stem cells,NSCs)移植联合PEP-1-SOD1对颅脑损伤后大鼠神经功能恢复的影响。方法:SD雄性大鼠80只,建立颅脑损伤模型后随机分为4组:对照组(损伤处注入生理盐水)-NSCs移植组(损伤处注入NSCs悬液)-SOD1组(损伤处注入PEP-1-SOD1蛋白)及NSCs+SOD1组(损伤处注入NSCs 悬液+PEP-1-SOD1蛋白)。4 d后采用定量PCR及Western blot法分别检测小鼠脑组织中AQP4基因表达及蛋白合成变化;分别于损伤后1-3 d和1-2-3-4周行Bederson评分,损伤后23~28 d行Morris水迷宫试验对大鼠运动神经功能进行评价;伤后第4周取材行病理切片BrdU 免疫组织化学染色。结果:脑损伤后4 d,对照组脑损伤周围组织AQP4及其mRNA的表达高于NSCs移植组及SOD1组,NSCs移植组及SOD1组均高于NSCs+SOD1组(P < 0.05)。分别于损伤后1-3 d及1-2-3-4周行Bederson评分显示4周时各组大鼠Bederson评分均低于24 h,并且4周时Bederson评分NSCs+SOD1组低于对照组(P < 0.05)。Morris 水迷宫试验结果显示NSCs+SOD1组神经功能改善较对照组明显(P <0.05)。免疫组化染色结果显示NSCs+SOD1组大鼠损伤灶脑组织中的BrdU 阳性细胞数高于NSCs移植组-SOD1组和对照组(P <0.05)。结论:神经干细胞移植可明显改善重型颅脑损伤后大鼠的神经学功能,联合应用PEP-1-SOD1有协同效果。

    Abstract:

    Objective:To investigate the effects of neural stem cell (NSC)transplantation combined with PEP-1-SOD1 on recovery of neurologic function of rats with traumatic brain injury. Methods:A total of 80 healthy SD rats were randomly divided into four groups including the control group (with injection of physiological saline into injury region),the NSCs transplantation group (suspension of NSCs),the SOD1 group (PEP-1-SOD1 protein),and the NSCs transplantation + SOD1 group (suspension of NSCs + PEP-1-SOD1 protein). AQP4 mRNA and AQP4 expressions were determined by quantitative polymerase chain reaction (QPCR)and Western blot assay at the 4th day after traumatic brain injury. Bederson neurobehavioral assessment was performed 24 hours,3 days,1,2,3 and 4 weeks after traumatic brain injury. Morris water maze test was performed to determine neurologic function of rats from day 23 to 28 after traumatic brain injury. Immunohistochemistry and pathology changes were determined at 4 weeks after injury. Results:The expressions of AQP4 mRNA and AQP4 in the control group were higher than those in the NSCs transplantation group and SOD1 group,and the expressions in the NSCs transplantation group and SOD1 group were higher than those in the NSCs transplantation+SOD1 group(P < 0.05). Bederson scores at 4 weeks after traumatic brain injury were lower than those at 24 hours in all groups. At 4 weeks after traumatic brain injury,the scores in the NSCs transplantation+SOD1 group were lower than those in the control group(P < 0.05). Morris water maze test showed that neurologic function in the NSCs+SOD1 group was significantly more improved than that in the control group (P < 0.05). Immunohistochemical staining showed that the number of BrdU positive cells in the NSCs transplantation+SOD1 group was higher than that in the control group,NSCs transplantation group,and SOD1 group(P < 0.05). Conclusion:PEP-1-SOD1 combined with transplantation of neural stem cells can significantly improve the neurological function in rats with traumatic brain injury.

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贾进明,陈菲菲,吴云飞,赵卫忠.神经干细胞移植联合PEP-1-SOD1对颅脑损伤大鼠神经功能恢复的影响[J].南京医科大学学报(自然科学版),2015,(11):1511-1516

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  • 收稿日期:2015-03-25
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  • 在线发布日期: 2015-11-30
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