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| 诱导型神经干细胞移植抑制颅脑创伤后补体活化的影响 | |
| 引用本文： | 高谋,徐如祥,王文佳,董勤,丁柏匀,姚慧,杨志军. 诱导型神经干细胞移植抑制颅脑创伤后补体活化的影响[J]. 中华神经创伤外科电子杂志, 2019, 5(1): 40-46. DOI: 10.3877/cma.j.issn.2095-9141.2019.01.009 |
| 作者姓名： | 高谋 徐如祥 王文佳 董勤 丁柏匀 姚慧 杨志军 |
| 作者单位： | 1. 100048 北京,解放军总医院第六医学中心神经外科2. 100700 北京,解放军总医院第七医学中心原附属八一脑科医院3. 572013 三亚,解放军总医院海南医院耳鼻咽喉头颈外科4. 100038 北京,首都医科大学附属复兴医院神经内科 |
| 基金项目： | 国家自然科学基金面上项目(81671189); 全军"十三五"军事医学创新工程项目(16CXZ001) |
| 摘 要： | 目的研究诱导型神经干细胞（iNSCs）移植对颅脑创伤（TBI）后补体活化的影响。 方法采用自由落体脑打击装置制备雄性成年C57BL/6小鼠TBI模型,将30只神经功能缺损评分（NSS）为4~8分的小鼠纳入TBI组,按照随机数字表法选取10只用于制备TBI小鼠血清和热灭活TBI小鼠血清,余下20只按照按照随机数字表法分为：iNSCs移植组（10只）和磷酸缓冲盐溶液（PBS）处理组（10只）。同时设置假手术（sham）组（10只）。于TBI后12 h,分别将含5×106个iNSCs单细胞悬液或等体积PBS经尾静脉注射到TBI小鼠体内,于移植后7 d处死动物,取脑组织行双重免疫荧光染色,观察iNSCs移植对TBI小鼠脑内C3d和NeuN抗体双阳性、C5b-9和NeuN抗体双阳性、C3d和Map2抗体双阳性以及C5b-9和Map2抗体双阳性的神经细胞的影响。于TBI后12 h,制备TBI小鼠血清和热灭活TBI小鼠血清,分别处理iNSCs后用流式细胞仪检测iNSCs表达补体调节分子Crry、Cd46、Cd59a和Cd55水平。 结果双重免疫荧光染色示：相比sham组,TBI小鼠脑组织中明显可见C3d和C5b-9沉积于NeuN和Map2抗体阳性的神经细胞;相比PBS处理组,iNSCs移植组TBI小鼠脑内C3d和NeuN抗体双阳性、C5b-9和NeuN抗体双阳性、C3d和Map2抗体双阳性以及C5b-9和Map2抗体双阳性的神经细胞数量均明显下降,2组差异具有统计学意义（P<0.05）。流式细胞仪检测示：相比热灭活TBI小鼠血清组,TBI小鼠血清组中iNSCs补体调节分子Crry表达水平明显上调,差异具有统计学意义（P<0.05）。 结论经静脉移植iNSCs可抑制TBI后补体活化,减轻神经损伤。 |
| 关 键 词： | 诱导型神经干细胞 颅脑创伤 补体活化 补体调节分子 移植 |
| 收稿时间： | 2018-11-11 |
Effects of induced neural stem cells on complement activation following traumatic brain injury |
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| Mou Gao,Ruxiang Xu,Wenjia Wang,Qin Dong,Boyun Ding,Hui Yao,Zhijun Yang. Effects of induced neural stem cells on complement activation following traumatic brain injury[J]. Chinese Journal of Neurotraumatic Surgery, 2019, 5(1): 40-46. DOI: 10.3877/cma.j.issn.2095-9141.2019.01.009 | |
| Authors: | Mou Gao Ruxiang Xu Wenjia Wang Qin Dong Boyun Ding Hui Yao Zhijun Yang |
| Affiliation: | 1. Department of Neurosurgery, The Sixth Medical Center, The PLA General Hospital, Beijing 100048, China2. Affiliated BaYi Brain Hospital, The Seventh Medical Center, The PLA General Hospital, Beijing 100700, China3. Department of ENT-HN, Hainan Hospital of PLA General Hospital, Sanya 572013, China4. Department of Neurology, Fuxing Hospital, Capital Medical University, Beijing 100038, China |
| Abstract: | ObjectiveTo study the effects of grafted induced neural stem cells (iNSCs) on complement activation following traumatic brain injury (TBI). MethodsHealthy adult male C57BL/6 mouse TBI models were established using a standardized weight-drop device. Neurological severity scores (NSS) of 4-8 points was included in TBI group (n=30). Ten TBI mice serum and heat-inactivated TBI mice serum were selected randomly, and the other 20 mice were randomly divided into iNSCs group (n=10) and phosphate buffer saline (PBS) group (n=10). At 12 h after TBI, mice were anaesthetized again and randomly selected to receive 200 μL of cell suspension containing 5×106 cells or PBS intravenously. On day 7 after TBI, animals were sacrificed for morphological analysis. Double-labeling experiments were utilized to determine the effects of iNSC grafts on C3d+/NeuN+, C5b-9+/NeuN+, C3d+/Map2+ and C5b-9+/Map2+ neurons in the brains of TBI mice. To identify the regulatory effects of iNSCs on complement activation, we utilized flow cytometry to determine Crry, Cd46, Cd59a and Cd55 protein expression levels in iNSCs after TBI or heat-inactivated TBI (HI-TBI) mouse serum treatment. ResultsDouble-labelling experiments showed that C3d and C5b-9 immunostaining was detected in the NeuN+ and Map2+ neurons in the injured cortex. Furthermore, the levels of C3d+/NeuN+, C5b-9+/NeuN+, C3d+/Map2+ and C5b-9+/Map2+ neurons were significantly lower in the iNSCs group than in the PBS group (P<0.05). Flow cytometry analysis revealed a dramatic increase in Crry expression in iNSCs in the TBI group compared with the HI-TBI group (P<0.05). ConclusionSystemic administration of iNSCs could efficiently attenuate the detrimental effects of complement activation on neurons following TBI. |
| Keywords: | Induced neural stem cell Traumatic brain injury Complement activation Crry Transplantation |
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