Epidermal growth factor receptor inhibitor ameliorates excessive astrogliosis and improves the regeneration microenvironment and functional recovery in adult rats following spinal cord injury.
Epidermal growth factor receptor inhibitor ameliorates excessive astrogliosis and improves the regeneration microenvironment and functional recovery in adult rats following spinal cord injury.
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表皮生长因子受体抑制剂改善成年大鼠脊髓损伤后过度星形胶质细胞增生并改善再生微环境和功能恢复
DOI：
10.1186/1742-2094-11-71
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发表时间：
2014-04-05
影响因子：
9.3
通讯作者：
Zou J
中科院分区：
文献类型：
--
作者：
Li ZW;Li JJ;Wang L;Zhang JP;Wu JJ;Mao XQ;Shi GF;Wang Q;Wang F;Zou J
研究背景星形胶质细胞增生是脊髓损伤（spinal cord injury，SCI）后的常见现象。虽然这一过程对轴突再生产生积极影响，但过度的星形胶质细胞增生对神经元的修复和恢复产生负面影响。表皮生长因子受体（EGFR）通路在反应性星形胶质细胞增生的调控中起关键作用，因此是更好地控制反应性星形胶质细胞增生的潜在治疗靶点。在这份报告中，我们的目的是调查是否使用EGFR酪氨酸激酶特异性抑制剂阻断EGFR信号可以减轻反应性星形胶质细胞增生和促进功能恢复后，创伤SCI.MethodThe星形胶质细胞划痕损伤模型在vitro和失重SCI模型在vivowere作为模型系统。PD 168393用于抑制EGFR信号传导激活。免疫荧光染色和Western blot分析星形胶质细胞活化和磷酸化EGFR（pEGFR）。通过免疫荧光检测位于伤口旁边的BrdU掺入细胞来确定增殖速率。ELISA法检测不同培养条件下培养液中TNF-α、iNOS、考克斯-2和IL-1β的含量。Western blot检测EGFR/pEGFR、胶质细胞酸性蛋白（GFAP）和硫酸软骨素蛋白聚糖（CSPGs）的表达。髓鞘用Luxol固蓝染色法染色。甲酚紫伊红染色分析损伤后损伤腔体积和神经元存活。最后，功能评分和残余尿记录进行显示大鼠' recovery.ResultsEGFR磷酸化被发现平行星形胶质细胞活化，和EGFR抑制剂PD 168393 potently抑制划痕诱导的反应性星形胶质细胞增生和促炎细胞因子/介质分泌的反应性星形胶质细胞在体外。此外，局部给药PD 168393在受伤的区域抑制CSPGs的生产和胶质瘢痕形成，并导致减少脱髓鞘和神经元丢失，这与显着的后肢运动功能和膀胱改善SCI rats.ConclusionsThe特异性EGFR抑制剂PD 168393可以改善过度反应性星形胶质细胞增生，促进SCI后轴突再生更有利的环境。因此，EGFR抑制剂可能是CNS损伤的有希望的治疗干预。
BackgroundAstrogliosis is a common phenomenon after spinal cord injury (SCI). Although this process exerts positive effects on axonal regeneration, excessive astrogliosis imparts negative effects on neuronal repair and recovery. Epidermal growth factor receptor (EGFR) pathway is critical to the regulation of reactive astrogliosis, and therefore is a potential target of therapeutics to better control the response. In this report, we aim to investigate whether blocking EGFR signaling using an EGFR tyrosine kinase specific inhibitor can attenuate reactive astrogliosis and promote functional recovery after a traumatic SCI.MethodThe astrocyte scratch injury modelin vitroand the weight-drop SCI modelin vivowere used as model systems. PD168393 was used to inhibit EGFR signaling activation. Astrocytic activation and phosphorylated EGFR (pEGFR) were observed after immunofluorescence staining and Western blot analysis. The rate of proliferation was determined by immunofluorescence detection of BrdU-incorporating cells located next to the wound. The levels of TNF-α, iNOS, COX-2 and IL-1β in the culture medium under different conditions were assayed by ELISA. Western blot was performed to semi-quantify the expression of EGFR/pEGFR, glial fibrillary acid protein (GFAP) and chondroitin sulfate proteoglycans (CSPGs). Myelin was stained by Luxol Fast Blue Staining. Cresyl violet eosin staining was performed to analyze the lesion cavity volume and neuronal survival following injury. Finally, functional scoring and residual urine recording were performed to show the rats’ recovery.ResultsEGFR phosphorylation was found to parallel astrocyte activation, and EGFR inhibitor PD168393 potently inhibited scratch-induced reactive astrogliosis and proinflammatory cytokine/mediator secretion of reactive astrocytesin vitro. Moreover, local administration of PD168393 in the injured area suppressed CSPGs production and glial scar formation, and resulted in reduced demyelination and neuronal loss, which correlated with remarkable hindlimb motor function and bladder improvement in SCI rats.ConclusionsThe specific EGFR inhibitor PD168393 can ameliorate excessive reactive astrogliosis and facilitate a more favorable environment for axonal regeneration after SCI. As such, EGFR inhibitor may be a promising therapeutic intervention in CNS injury.
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DOI：
10.1083/jcb.85.3.890
发表时间：
1980-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
McCarthy KD;de Vellis J
通讯作者：
de Vellis J
影响因子：
5.3
作者：
Jones, LL;Margolis, RU;Tuszynski, MH
通讯作者：
Tuszynski, MH
影响因子：
3.5
作者：
Evans, M. C.;Couch, Y.;Turner, M. R.
通讯作者：
Turner, M. R.
影响因子：
3.4
作者：
Davies, JE;Tang, XF;Davies, SJA
通讯作者：
Davies, SJA
影响因子：
6.2
作者：
Cua, Rowena C.;Lau, Lorraine W.;Yong, V. Wee
通讯作者：
Yong, V. Wee