Fasudil attenuates glial cell-mediated neuroinflammation via ERK1/2 and AKT signaling pathways after optic nerve crush
Fasudil attenuates glial cell-mediated neuroinflammation via ERK1/2 and AKT signaling pathways after optic nerve crush
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Fasudil 通过 ERK1/2 和 AKT 信号通路减轻视神经挤压后神经胶质细胞介导的神经炎症
DOI：
10.1007/s11033-020-05953-y
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发表时间：
2020-11-08
影响因子：
2.8
通讯作者：
Cui, Ling
中科院分区：
文献类型：
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作者：
Huang, Wei;Lan, Qianqian;Cui, Ling
研究法舒地尔在视神经夹伤（ONC）中的作用，并进一步探索其可能的分子机制。ONC损伤后，大鼠每天腹腔注射一次法舒地尔或生理盐水，直至处死。通过荧光金逆行标记评估视网膜神经节细胞（RGCs）的存活情况。通过免疫荧光检测视网膜神经胶质细胞的活化和数量变化（胶质纤维酸性蛋白GFAP、离子钙结合衔接分子1 iba - 1）。通过蛋白质印迹法检测裂解的半胱天冬酶3和9、磷酸化细胞外调节蛋白激酶1/2（p - ERK1/2）和磷酸化蛋白激酶B（p - AKT）的表达。采用实时聚合酶链反应测定促炎细胞因子的水平。法舒地尔治疗抑制了RGCs凋亡，减少了RGCs损失，这分别通过ONC后凋亡相关蛋白表达降低以及RGCs的荧光金标记增加得以证明。此外，与ONC组相比，ONC +法舒地尔组的GFAP和iba1表达显著降低。ONC +法舒地尔组的促炎细胞因子水平明显低于ONC组。此外，法舒地尔治疗显著提高了ERK1/2和AKT的磷酸化水平（p - ERK1/2和p - AKT）。我们的研究表明，在大鼠ONC模型中法舒地尔通过上调ERK1/2和AKT信号通路减轻神经胶质细胞介导的神经炎症。我们得出结论，法舒地尔可能是创伤性视神经病变的一种新型治疗方法。
To investigate the functional role of fasudil in optic nerve crush (ONC), and further explore its possible molecular mechanism. After ONC injury, the rats were injected intraperitoneally either with fasudil or normal saline once a day until euthanized. RGCs survival was assessed by retrograde labeling with FluoroGold. Retinal glial cells activation and population changes (GFAP, iba-1) were measured by immunofluorescence. The expressions of cleaved caspase 3 and 9, p-ERK1/2 and p-AKT were detected by western blot. The levels of the pro-inflammatory cytokines were determined using real-time polymerase chain reaction. Fasudil treatment inhibited RGCs apoptosis and reduced RGCs loss demonstrated by the decreased apoptosis-associated proteins expression and the increased fluorogold labeling of RGCs after ONC, respectively. In addition, the ONC + fasudil group compared had a significantly lower expression of GFAP and iba1 compared with the ONC group. The levels of pro-inflammatory cytokines were significantly reduced in the ONC + fasudil group than in the ONC group. Furthermore, the phosphorylation levels of ERK1/2 and AKT (p-ERK1/2 and p-AKT) were obviously elevated by the fasudil treatment. Our study demonstrated that fasudil attenuated glial cell-mediated neuroinflammation by up-regulating the ERK1/2 and AKT signaling pathways in rats ONC models. We conclude that fasudil may be a novel treatment for traumatic optic neuropathy.