Progranulin deficiency exacerbates spinal cord injury by promoting neuroinflammation and cell apoptosis in mice
Progranulin deficiency exacerbates spinal cord injury by promoting neuroinflammation and cell apoptosis in mice
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DOI：
10.1186/s12974-019-1630-1
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发表时间：
2019-11-27
影响因子：
9.3
通讯作者：
Liu, Chuan-ju
中科院分区：
文献类型：
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作者：
Wang, Chao;Zhang, Lu;Liu, Chuan-ju
目的脊髓损伤（SCI）常导致严重的灾难性功能障碍和残疾，给社会造成巨大的经济负担。本研究旨在确定前颗粒蛋白（PGRN）是否在脊髓损伤后的进行性损伤中发挥作用，并评估PGRN衍生物作为脊髓损伤新治疗靶点的发展潜力。方法采用失重法对pgrn缺陷（Gr(-/-)）和野生型（WT）仔鼠进行脊髓损伤实验。Western blotting和免疫荧光法检测损伤后局部PGRN表达。在指定时间点进行Basso小鼠量表（BMS）、倾斜网格行走测试和斜面测试，评估神经功能恢复情况。采用组织学（苏木精和伊红（H&E）染色、尼氏染色、TUNEL法和免疫荧光法）、Western blotting（全组织蛋白检测iNOS/p-p65/Bax/Bcl-2）和离体ELISA（检测TNF α /IL-1 β /IL-6/IL-10）检测炎症和细胞凋亡。为了确定靶向PGRN的预防和治疗潜力，将PGRN衍生的小蛋白atstrin与PLGA-PEG-PLGA热敏水凝胶偶联，并在脊髓损伤前注射到鞘内间隙。记录BMS记录神经恢复情况，Western blotting检测炎症和凋亡蛋白。结果脊髓损伤后，PGRN在活化的巨噬细胞/小胶质细胞中高表达，并在损伤后第7天达到峰值。与WT对照组相比，Grn（-/-）小鼠在损伤后第21、28、35和42天表现出延迟的神经恢复。组织学、TUNEL实验、免疫荧光、Western blotting和ELISA均显示Grn（-/-）小鼠表现出不受控制和扩大的炎症和细胞凋亡。控释阿司特林可改善PGRN缺乏的神经功能恢复和促炎/促凋亡作用。结论PGRN缺乏通过促进神经炎症和细胞凋亡而加重脊髓损伤，阿司德林可减轻这一作用。总的来说，我们的数据为使用PGRN衍生物作为一种有希望的治疗方法来改善脊髓损伤患者的功能恢复提供了新的证据。
Purpose Spinal cord injury (SCI) often results in significant and catastrophic dysfunction and disability and imposes a huge economic burden on society. This study aimed to determine whether progranulin (PGRN) plays a role in the progressive damage following SCI and evaluate the potential for development of a PGRN derivative as a new therapeutic target in SCI. Methods PGRN-deficient (Gr(-/-)) and wild-type (WT) littermate mice were subjected to SCI using a weight-drop technique. Local PGRN expression following injury was evaluated by Western blotting and immunofluorescence. Basso Mouse Scale (BMS), inclined grid walking test, and inclined plane test were conducted at indicated time points to assess neurological recovery. Inflammation and apoptosis were examined by histology (Hematoxylin and Eosin (H&E) staining and Nissl staining, TUNEL assays, and immunofluorescence), Western blotting (from whole tissue protein for iNOS/p-p65/Bax/Bcl-2), and ex vivo ELISA (for TNF alpha/IL-1 beta/IL-6/IL-10). To identify the prophylactic and therapeutic potential of targeting PGRN, a PGRN derived small protein, Atsttrin, was conjugated to PLGA-PEG-PLGA thermosensitive hydrogel and injected into intrathecal space prior to SCI. BMS was recorded for neurological recovery and Western blotting was applied to detect the inflammatory and apoptotic proteins. Results After SCI, PGRN was highly expressed in activated macrophage/microglia and peaked at day 7 post-injury. Grn(-/-) mice showed a delayed neurological recovery after SCI at day 21, 28, 35, and 42 post-injury relative to WT controls. Histology, TUNEL assay, immunofluorescence, Western blotting, and ELISA all indicated that Grn(-/-) mice manifested uncontrolled and expanded inflammation and apoptosis. Administration of control-released Atsttrin could improve the neurological recovery and the pro-inflammatory/pro-apoptotic effect of PGRN deficiency. Conclusion PGRN deficiency exacerbates SCI by promoting neuroinflammation and cellular apoptosis, which can be alleviated by Atsttrin. Collectively, our data provide novel evidence of using PGRN derivatives as a promising therapeutic approach to improve the functional recovery for patients with spinal cord injury.