天津医药

天津医药 ›› 2018, Vol. 46 ›› Issue (4): 350-356.doi: 10.11958/20171090

• 实验研究 • 上一篇    下一篇

脂肪间充质干细胞分泌组对颅脑创伤后 大鼠脑水肿的影响

徐超 1，李晓红 1，史洪建 2，王晶 1，王丽娜 1，符锋 1，赵万勇 1，朱旭 1，孙洪涛 1，涂悦 1，栾佐 3，张赛 1△   

  1. 基金项目：国家自然科学基金项目资助（81471275，81771352，81671222，81771350）；天津市自然科学基金项目资助（16JCYBJC27600） 作者单位：1中国人民武装警察部队后勤学院附属医院颅脑创伤与神经修复研究所（邮编300162）；2中国人民武装警察部队后勤学院附 属医院核磁共振科；3中国人民解放军海军总医院儿科 作者简介：徐超（1991），男，硕士在读，主要从事颅脑创伤相关研究 △通讯作者 E-mail：zhangsai718@vip.126.com
  • 收稿日期:2017-10-16 修回日期:2018-02-25 出版日期:2018-04-15 发布日期:2018-04-15
  • 通讯作者: 张赛 E-mail:zhangsai718@vip.126.com
  • 基金资助:
    国家自然科学基金;天津市自然科学基金

The effect of the secretome of adipose derived from mesenchymal stem cells on cerebral edema of rats after traumatic brain injury

XU Chao1, LI Xiao-hong1, SHI Hong-jian2, WANG Jing1, WANG Li-na1, FU Feng1, ZHAO Wan-yong1, ZHU Xu1, SUN Hong-tao1, TU Yue1, LUAN Zuo3, ZHANG Sai1△   

  1. 1 Institution of Brain Trauma and Neurology Disease of the Affiliated Hospital of Logistics University of PAP, Tianjin 300162, China; 2 Department of MRI of the Affiliated Hospital of Logistics University of PAP; 3 Department of Pediatrics of General Hospital of Navy, PLA △Corresponding Author E-mail：zhangsai718@vip.126.com
  • Received:2017-10-16 Revised:2018-02-25 Published:2018-04-15 Online:2018-04-15

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摘要： 摘要：目的 探讨脂肪间充质干细胞分泌组（ASC-ST）对大鼠颅脑创伤（TBI）后继发脑水肿的影响及其潜在机 制。方法 将 70 只 SD 大鼠随机分为 Sham 组（n=18）、TBI 组（n=26）和 TBI+ST 组（n=26）。应用电子颅脑损伤仪 （eCCI）建立TBI大鼠模型，Sham组只开骨窗；TBI组经尾静脉注射0.3 mL生理盐水，TBI+ST组经尾静脉注射0.2 mL ASC-ST 和 0.1 mL 生理盐水，连续注射 7 d。在 TBI 后 3、7、14 和 21 d 对 TBI 和 TBI+ST 组 8 只大鼠行神经功能评分 （mNSS）；每组选取6只大鼠在TBI后3、7和14 d行头颅核磁（MRI）测量表观弥散系数（ADC）；采用干湿比重法测量脑 水含量并提取损伤周围脑组织总RNA，采用实时定量多聚酶链反应（qRT-PCR）检测肿瘤坏死因子（TNF）-α、白细胞 介素（IL）-1β和IL-6的mRNA 表达水平。结果 TBI 后14、21 d时，TBI+ST 组mNSS 评分均低于TBI 组（P＜0.05）。 TBI后3 d，TBI组和TBI+ST组损伤周围皮层（IC）和损伤对侧皮层（CC）的ADC值均高于Sham组（P＜0.05）；TBI后7 d，TBI+ST组IC和损伤侧海马（IH）的ADC值均低于TBI组（P＜0.05）；TBI后14 d，TBI+ST组IC和CC的ADC值均低于 TBI组（P＜0.05）。TBI后3 d和7 d，TBI+ST组大鼠脑组织水含量与TBI组差异无统计学意义（P＞0.05）；TBI后14 d， TBI+ST组大鼠脑组织水含量低于TBI组（P＜0.05）。TBI后3 d，Sham组大鼠损伤周围脑组织中IL-6表达显著低于 TBI组和TBI+ST组，TBI+ST组TNF-α表达显著低于Sham组和TBI组，TBI组高于Sham组；TBI+ST组与TBI组IL-1β 表达均低于Sham组（均P＜0.05）。TBI后7 d，TBI+ST组与Sham组IL-6表达均低于TBI组，TBI+ST组低于Sham组； TBI组TNF-α表达高于Sham组；TBI组IL-1β表达高于其余两组，Sham组高于TBI+ST组（均P＜0.05）。TBI后14 d， TBI+ST组与Sham组IL-6表达均低于TBI组；TBI+ST组TNF-α表达仍低于其他组；TBI组IL-1β表达仍高于其余两组 （均P＜0.05）。结论 ASC-ST可通过减少TBI后炎症反应和炎性因子表达，显著改善大鼠TBI后脑水肿状况和神经 功能预后，是一种具有较高临床推广价值的生物治疗药物。

关键词: 间质干细胞, 脑损伤, 脑水肿, 干细胞分泌组, 颅脑创伤, 炎症反应

Abstract: Abstract：Objective To investigate the effect of adipose-derived mesenchymal stem cells (ASC-ST) on the secondary brain edema of rats after traumatic brain injury (TBI) and its potential mechanisms. Methods Seventy SD rats were randomly divided into three groups: Sham group (n=18), TBI group (n=26) and TBI + ST group (n=26). TBI models were established by electric cortical contusion impactor (eCCI). Rats from sham group only received the craniectomy without impact. Rats from TBI + ST group were injected with 0.2 mL ASC-ST and 0.1 mL normal saline, while rats from TBI group were injected with 0.3 mL normal saline through tail veins after TBI. The injection was persisted for 7 days. Modified neurological function scores (mNSS) were performed on 8 rats from TBI and TBI+ST groups at 3, 7 and 14 days after TBI. Six rats of each groups accepted magnetic resonance imaging (MRI) at 3, 7 and 14 days after TBI. The water content of brain was measured, and total RNA was exacted from brain tissue of the injured lesions. The mRNA expression levels of TNF-α, IL- 1β and IL-6 were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Results At 14 and 21 days after TBI, rats from TBI + ST group got lower mNSS scores than those from TBI group (P＜0.05). At 3 days after TBI, compared with those in Sham group, the ADC values of ipsilateral cortex (IC) and contralateral cortex (CC) were higher in both TBI and TBI+ST groups (P＜0.05). At 7 days after TBI, the ADC values of IC and ipsilateral hippocampus (IH) were lower in TBI + ST group than those in TBI group (P＜0.05). At 14 days after TBI, ADC values of IC and CC were lower in TBI + ST group than those in TBI group (P＜0.05). At 3 and 7 days after TBI, the water content of brain tissue of TBI group was similar with TBI+ST group (P＞0.05). At 14 days after TBI, the water content of brain tissue was lower in TBI+ST group than that of TBI group (P＜0.05). At 3 days after TBI, the expression of IL-6 was extremely lower in Sham group than that in TBI and TBI+ST groups, while expression of TNF-α was higher in TBI+ST group than that in TBI and Sham groups, and expression of TNF-α was higher in TBI group than that of Sham group (P＜0.05). At 7 days after TBI, expressed levels of IL- 6 were lower in TBI+ST and Sham groups than those of TBI group, and the expression was lower in TBI+ST group than that in Sham group; The more mRNA expression of TNF-α was found in TBI group than Sham group, and more IL-1β than other groups; The more IL-1β expression was found in Sham group than TBI+ST group (all P＜0.05). At 14 days after TBI, both TBI+ST and Sham groups were found less mRNA of IL-6 than TBI group, TBI+ST group was found less TNF-α expression than other two groups, but the expression of IL-1β in TBI group was still higher than that in other groups (all P＜0.05). Conclusion ASC-ST can significantly reduce the brain edema of rats after TBI and improve the prognosis of neurological function by mitigating the inflammatory responses. It is a promising kind of biotherapeutic drug with high clinical value.

Key words: mesenchymal stem cells, brain injuries, brain edema, secretome of stem cells, traumatic brain injury, inflammation