中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (10): 1759-1770.doi: 10.4103/1673-5374.238617

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

亚低温联合脂肪干细胞移植治疗缺血性脑损伤

  

  • 收稿日期:2018-07-20 出版日期:2018-10-15 发布日期:2018-10-15
  • 基金资助:

    国家自然科学基金（81371301）

Combination of mild therapeutic hypothermia and adipose-derived stem cells for ischemic brain injury

Kai Zhao1, Rui Li2, Sheng Bi3, Yu Li1, Long Liu1, Yu-Long Jia1, Peng Han1, Chang-Cong Gu4, Xi-Ze Guo4, Wan-Ping Zhang4, Chun Wang4, Chun-Ying Pei4, Lin-Lu Tian4, Li-Xian Li1   

  1. 1 Department of Neurosurgery, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, China
    2 Department of Neurology, University of Pennsylvania, Philadelphia, PA, USA
    3 Department of Neurology, First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, China
    4 Department of Immunology, Harbin Medical University, Harbin, Heilongjiang Province, China
  • Received:2018-07-20 Online:2018-10-15 Published:2018-10-15
  • Contact: Li-Xian Li, MD,lilixian@hrbmu.edu.cn.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81371301.

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摘要：

亚低温治疗可减轻脑缺血，降低脑水肿，改善脑缺血患者的预后。脂肪来源间充质干细胞移植治疗在脑缺血时也可减少梗塞区域神经元死亡及炎细胞的浸润，从而发挥神经保护作用。实验设想亚低温和脂肪干细胞移植联合应用治疗缺血性脑损伤可取得更好的神经保护效果，通过线栓法构建短暂性大脑中动脉闭塞模型，在脑缺血2 h期间给予亚低温（33℃）治疗，于再灌注时通过股静脉给予脂肪干细胞移植治疗。通过mNSS评分对模型大鼠神经功能缺陷进行评估，通过TTC染色检测梗死区域的变化，通过TUNEL染色检测脑组织凋亡细胞的变化，通过免疫荧光染色检测脑组织微血管的再生以及胶质瘢痕的变化，通过对缺血部位的组织学染色来检测脑缺血后的炎细胞浸润情况，通过RT-PCR来检测缺血脑组织炎性细胞因子mRNA的表达。结果发现，与单一的亚低温治疗组和脂肪干细胞移植治疗组相比，两者联合治疗能更有效的改善神经功能缺陷，减少梗死区域，可显著降低体内TUNEL阳性细胞数量，减少星形胶质细胞标志物GFAP阳性细胞数量，提高微血管的再生标志物VEGF免疫反应，有效减少炎性细胞浸润，下调促炎因子IL-1β、TNFa和IL-6 mRNA的表达。我们的数据表明，亚低温联合脂肪干细胞移植治疗缺血性脑损伤，效果优于两种方法单独治疗。

orcid:0000-0003-4353-2229(Li-Xian Li)

 

关键词: 脑损伤, 脑卒中, 大鼠, 短暂性大脑中动脉闭塞, 脑复苏, 亚低温治疗, 脂肪干细胞, 联合治疗, 神经保护, 神经细胞死亡, 神经再生

Abstract:

Mild therapeutic hypothermia has been shown to mitigate cerebral ischemia, reduce cerebral edema, and improve the prognosis of patients with cerebral ischemia. Adipose-derived stem cell-based therapy can decrease neuronal death and infiltration of inflammatory cells,exerting a neuroprotective effect. We hypothesized that the combination of mild therapeutic hypothermia and adipose-derived stem cells would be neuroprotective for treatment of stroke. A rat model of transient middle cerebral artery occlusion was established using the nylon monofilament method. Mild therapeutic hypothermia (33°C) was induced after 2 hours of ischemia. Adipose-derived stem cells were administered through the femoral vein during reperfusion. The severity of neurological dysfunction was measured by a modified Neurological Severity Score Scaling System. The area of the infarct lesion was determined by 2,3,5-triphenyltetrazolium chloride staining.Apoptotic neurons were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining.The regeneration of microvessels and changes in the glial scar were detected by immunofluorescence staining. The inflammatory responses after ischemic brain injury were evaluated by in situ staining using markers of inflammatory cells. The expression of inflammatory cytokines was measured by reverse transcription-polymerase chain reaction. Compared with mild therapeutic hypothermia or adipose-derived stem cell treatment alone, their combination substantially improved neurological deficits and decreased infarct size. They synergistically reduced the number of TUNEL-positive cells and glial fibrillary acidic protein expression, increased vascular endothelial growth factor levels, effectively reduced inflammatory cell infiltration and down-regulated the mRNA expression of the proinflammatory cytokines interleukin-1β, tumor necrosis factor-α and interleukin-6. Our findings indicate that combined treatment is a better approach for treating stroke compared with mild therapeutic hypothermia or adipose-derived stem cells alone.

Key words: nerve regeneration, brain injury, stroke, rats, transient middle cerebrum artery occlusion, cerebral resuscitation, mild therapeutic hypothermia, adipose-derived stem cells, combination therapy, neuroprotection, neuronal cell death, neural regeneratio