Inhibition of NOX2 signaling limits pain-related behavior and improves motor function in male mice after spinal cord injury: Participation of IL-10/miR-155 pathways
Inhibition of NOX2 signaling limits pain-related behavior and improves motor function in male mice after spinal cord injury: Participation of IL-10/miR-155 pathways
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DOI：
10.1016/j.bbi.2019.02.024
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发表时间：
2019-08
期刊：
影响因子：
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通讯作者：
B. Sabirzhanov;Yun Li;Marino Coll-Miro;Jessica J. Matyas;Junyun He;Alok Kumar;N. Ward;Jingwen Yu
中科院分区：
文献类型：
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作者：
B. Sabirzhanov;Yun Li;Marino Coll-Miro;Jessica J. Matyas;Junyun He;Alok Kumar;N. Ward;Jingwen Yu
NADPH氧化酶（NOX 2）是一种诱导活性氧（ROS）的酶，并作为促炎和神经恢复性小胶质细胞/巨噬细胞表型之间的开关;这种变化在神经性疼痛和运动功能障碍中起重要作用。据报道，脊髓损伤（SCI）后NOX 2表达增加，抑制NOX 2可改善运动功能。然而，NOX 2在创伤后疼痛和运动障碍中的潜在机制仍未被探索。在本研究中，我们报告了NOX 2（NOX 2 −/−）的耗竭或使用NOX 2ds-tat抑制NOX 2显著降低了雄性小鼠在T10中度挫伤SCI后的机械/热皮肤超敏反应和运动功能障碍。Western blot（WB，3 mm病变面积）和免疫组织化学（IHC）显示，SCI主要在损伤后8周内在小胶质细胞/巨噬细胞中升高NOX 2表达。在损伤区域和腰椎增大中，通过流式细胞术检测到的损伤后24小时的NOX 2缺失显著减少了CD 11b +/CD 45 hiF 4/80+巨噬细胞浸润，并且通过IHC检测到损伤后8周的8-OHG+ROS产生。NOX 2缺乏也改变了小胶质细胞/巨噬细胞促炎和抗炎平衡对神经恢复反应。WB分析显示，NOX 2 −/−小鼠中精氨酸酶-1和YM 1蛋白的显著增加。此外，qPCR分析显示，NOX 2 −/−小鼠中抗炎细胞因子IL-10水平显著上调，与microRNA-155表达减少相关。这些发现在损伤后3天从脊髓分离的CD 11b+小胶质细胞/巨噬细胞中得到证实。总而言之，我们的数据表明IL-10/miR-155途径在调节NOX 2介导的SCI功能障碍中发挥重要作用。因此，特异性靶向NOX 2可能为治疗SCI患者的神经功能障碍提供有效的策略。
NADPH oxidase (NOX2) is an enzyme that induces reactive oxygen species (ROS) and serves as a switch between the pro-inflammatory and neurorestorative microglial/macrophage phenotypes; such changes play an important role in neuropathic pain and motor dysfunction. Increased NOX2 expression after spinal cord injury (SCI) has been reported, and inhibition of NOX2 improves motor function. However, the underlying mechanisms of NOX2 in post-traumatic pain and motor deficit remain unexplored. In the present study, we report that depletion of NOX2 (NOX2−/−) or inhibition of NOX2 using NOX2ds-tat significantly reduced mechanical/thermal cutaneous hypersensitivity and motor dysfunction after moderate contusion SCI at T10 in male mice. Western blot (WB, 3 mm lesion area) and immunohistochemistry (IHC) showed that SCI elevates NOX2 expression predominantly in microglia/macrophages up to 8 weeks post-injury. Deletion of NOX2 significantly reduced CD11b+/CD45hiF4/80+macrophage infiltration at 24 h post-injury detected by flow cytometry and 8-OHG+ROS production at 8 weeks post-injury by IHC in both lesion area and lumbar enlargement. NOX2 deficiency also altered microglial/macrophage pro-inflammatory and anti-inflammatory balance towards the neurorestorative response. WB analysis showed robust increase of Arginase-1 and YM1 proteins in NOX2−/−mice. Furthermore, qPCR analysis showed significant up-regulation of anti-inflammatory cytokine IL-10 levels in NOX2−/−mice, associated with reduced microRNA-155 expression. These findings were confirmed in CD11b+microglia/macrophages isolated from spinal cord at 3 days post-injury. Taken together, our data suggest an important role for IL-10/miR-155 pathway in regulating NOX2-mediated SCI-dysfunction. Thus, specific targeting of NOX2 may provide an effective strategy for treating neurological dysfunction in SCI patients.