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小鼠蛛网膜下腔出血后早期脑损伤期转运蛋白的表达

Expression of Translocator Protein in Early Brain Injury After Subarachnoid Hemorrhage in Mice

    摘要
  • 摘要:
      目的  评估小鼠蛛网膜下腔出血(subarachnoid hemorrhage，SAH)后72 h内(早期脑损伤期)脑组织中转运蛋白(translocator protein，TSPO)的表达。
      方法  将44只C57BL/6J小鼠随机分成两组，Sham组17只，SAH组27只。SAH组通过血管内穿刺法构建SAH模型，Sham组不刺破血管。造模前(0 h)，造模后6 h、24 h、48 h、72 h，两组采用改良加西亚评分对小鼠进行神经功能评分，造模后各时点处死SAH组小鼠(Sham组仅取造模后24 h处死)，以Western blot、正电子发射计算机断层显像(positron emission tomography-computed tomography，PET-CT)和免疫荧光染色评价TSPO在脑组织中的表达，免疫荧光染色评估TSPO与小胶质细胞的共定位。
      结果  SAH组小鼠神经功能评分造模后随时间先降低后增加，脑组织中TSPO表达量造模后随时间先增加后降低，两者呈负相关(r=-0.615 6，P < 0.01)。PET-CT提示SAH后小鼠脑组织示踪剂摄取量较Sham组升高，差异有统计学意义(P < 0.05)。免疫荧光染色提示SAH组出血侧顶叶皮层和基底皮层均有TSPO表达增加，且与小胶质细胞标记物Iba-1共定位。
      结论  SAH后早期脑损伤期，TSPO在脑组织中的表达部位广泛增加，表达量呈现先增加后减少的趋势，可能参与小胶质细胞的激活，并调控SAH后脑损伤的发生与发展。

     

    Abstract:
      Objective  To evaluate the expression of translocator protein (TSPO) in brain tissue within 72 h after subarachnoid hemorrhage (SAH) in mice.
      Methods  Forty-four C57BL/6J mice were randomly divided into two groups, 17 in the Sham group and 27 in the SAH group. SAH mice model was performed by endovascular perforation as previously described with slight modifications. Sham group mice were performed by the same method but without piercing the blood vessels. Before and 6 h, 24 h, 48 h, 72 h after modeling, the two groups were scored with modified Garcia score for neurological function. At 6 h, 24 h, 48 h, 72 h after modeling, the mice were sacrificed. Sham group mice were sacrificed at 24 h after modeling. The expression of TSPO in brain tissue was evaluated by Western blot, positron emission tomography-computed tomography (PET-CT) and immunofluorescence staining. Fluorescent double staining was used to assess the relationship of TSPO and microglia.
      Results  The neurological function scores of the SAH group mice decreased with time and then increased. The expression of TSPO in the brain tissue increased first and then decreased with time, and there was a negative correlation between them (r=-0.615 6, P < 0.01). PET-CT showed that the tracer intake of mouse brain tissue after SAH was higher than that of Sham group, and the difference was statistically significant (P < 0.05). Immunofluorescence staining showed that TSPO increased in the parietal cortex and basal cortex of the SAH group. And fluorescent double staining suggested that TSPO colocalized with Iba-1 which was a specific marker of microglia.
      Conclusion  In the early brain injury after SAH, the expression of TSPO in brain is widely increased, and the expression level increases first and then decreases. TSPO could participate in the activation of microglia and regulate the occurrence and development of brain injury after SAH.

     

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