姜黄素诱导内源性神经干细胞促进大鼠脊髓损伤后功能修复  

Curcumin induces endogenous neural stem cells to promote functional recovery in rats after spinal cord injury

作　　者：向鑫[1] 袁继超[1] 陈飞[1] 朱海涛[1] 储卫华[1] 林江凯[1] 

机构地区：[1]第三军医大学西南医院神经外科,全军神经外科研究所,全军神经创伤防治重点实验室,重庆400038

出　　处：《第三军医大学学报》2014年第9期883-887,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(81070979;81000531)~~

摘　　要：目的 探讨姜黄素对脊髓损伤后内源性神经干细胞分化的影响及其对损伤后功能修复的作用。 方法 雄性SD大鼠72只按随机数字表法分为3组：假手术组（n=12）、单纯损伤组（n=30）、姜黄素处理组（n=30）。假手术组仅行椎板切除术，单纯损伤组、姜黄素处理组以70 g力的动脉瘤夹钳夹T9-T10节段脊髓1 min造模。姜黄素处理组于脊髓损伤后30 min按200 mg/kg给予腹腔注射姜黄素，单纯损伤组给予等量生理盐水。分别于损伤后1 d，1、2、4、8周行BBB运动功能评分及斜板实验评价大鼠后肢功能，后处死大鼠并取材，行免疫荧光染色分别观察Nestin、Tuj1、GFAP/CSPG的表达，Western blot检测Wnt3a蛋白及β-catenin表达情况。 结果 BBB运动功能评分及斜板实验结果显示损伤后自1周开始，各时间点姜黄素处理组大鼠后肢功能恢复情况均优于单纯损伤组（P〈0.05）。免疫荧光染色显示脊髓损伤后1周可见损伤区附近有Nestin阳性细胞，2周时达高峰，4周时明显减少，单纯损伤组、姜黄素处理组组间差异无统计学意义；姜黄素处理组Tuj1阳性神经元数量较单纯损伤组明显增多；4周时脊髓损伤中心囊腔周边形成胶质瘢痕，姜黄素处理组GFAP及CSPG表达明显弱于单纯损伤组。Western blot检测Wnt3a及β-catenin表达显示假手术组几乎无表达，姜黄素处理组表达量较单纯损伤组明显增强（P〈0.05）。 结论 姜黄素可通过激活Wnt/β-catenin信号通路诱导脊髓损伤后被激活的内源性神经干细胞更多地分化为神经元，减轻胶质瘢痕，从而改善大鼠后肢功能的恢复。Objective To determine the effect of curcumin on the differentiation of endogenous neural stem cells （ENSCs） and functional recovery after spinal cord injury （SCI） in rats. Methods Seventy-two male Sprague-Dawley （SD） rats were randomly divided into 3 groups, that is, sham-operation group （group A, n=12）, simple injury group （group B, n=30）, and curcumin treatment group （group C, n=30）. The rats of group A received only laminectomy. While, those of groups B and C were introduced 70 g closing force aneurysm clip injury at T9-T10 segment of spinal cord for 1 min, and the rats of group C was given 200 mg/kg curcumin intraperitoneally in 30 min after SCI but those of group B were given equivalent amount of normal saline. The Basso, Beattie and Bresnahan （BBB） locomotor rating scale and inclined plane test were used to evaluate rat behavioral consequences in 1 d, and 1, 2, 4 and 8 weeks after injury. Then the rats were sacrificed and the spinal cord tissue samples were harvested to observe the expression of Nestin, Tuj1, and GFAP/CSPG by immunofluorescence assay. The protein expression of Wnt3a and β-catenin was determined by Western blotting. Results BBB locomotor rating scale and inclined plane test showed that the functional recovery was significantly better in group C than group B at each time point from 1week after SCI （P〈0.05）. Immunofluorescence assay showed that Nestin positive cells were seen at the vicinity of the lesion site in 1 week after SCI, reached peak at 2 weeks, and obviously decreased at 4 weeks, but there was no significant difference between groups B and C. There were significant more Tuj1-labeled neurons in group C than in group B at each time point. The formation of glia scar surrounding the cavity at the center of lesion site was defined at 4 week after SCI, while the expression of GFAP and CSPG was significant decreased in group C than in group B. Western blotting showed that there was almost no expression of Wnt3a and β-catenin in group A, and th

关 键 词：姜黄素 脊髓损伤 内源性神经干细胞 WNT Β-CATENIN信号通路 

分 类 号：R282.71[医药卫生—中药学] R363.22[医药卫生—中医学]