RNAi-mediated ephrin-B2 silencing attenuates astroglial-fibrotic scar formation and improves spinal cord axon growth
RNAi-mediated ephrin-B2 silencing attenuates astroglial-fibrotic scar formation and improves spinal cord axon growth
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RNAi介导的ephrin-B2沉默减弱星形胶质细胞纤维化疤痕形成并改善脊髓轴突生长
DOI：
10.1111/cns.12723
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发表时间：
2017-10-01
影响因子：
5.5
通讯作者：
Wang, Xiao-Dong
中科院分区：
文献类型：
--
作者：
Li, Yi;Chen, Ying;Wang, Xiao-Dong
目的：中枢神经系统损伤后星形胶质细胞纤维化疤痕的形成有助于修复血脑屏障并封闭病变，同时它也为轴突再生提供了强大的屏障。为了消除/减少抑制部分，同时保留星形胶质纤维化疤痕的有益作用，我们进行了大量的临床前研究。方法：在本研究中，我们建立了一个体外系统，其中星形胶质细胞和脑膜成纤维细胞共培养，用外源性转化生长因子-β1（TGF-β1）处理，形成星形胶质细胞纤维化疤痕样细胞簇，从而评估RNAi靶向的功效ephrin-B2 从一开始就防止疤痕形成。我们在定制的微流体平台上进一步测试了基于 RNAi 的缓解星形胶质纤维化疤痕对脊髓轴突生长的效果。结果：我们发现，针对 ephrin-B2 的 siRNA 显着减少了 TGF-β1 诱导的细胞簇的数量和直径，并减少了共培养物中聚集蛋白聚糖和多功能蛋白聚糖的表达，并允许向外生长的脊髓轴突显着更长地延伸到星形胶质纤维化疤痕中结论：这些结果表明，肝配蛋白-B2 特异性 siRNA 可以减轻星形胶质细胞纤维化疤痕的形成，特别是聚集蛋白聚糖和多功能蛋白聚糖的表达，从而改善脊髓轴突再生的微环境。
Aims: Astroglial-fibrotic scar formation following central nervous system injury can help repair blood-brain barrier and seal the lesion, whereas it also represents a strong barrier for axonal regeneration. Intensive preclinical efforts have been made to eliminate/reduce the inhibitory part and, in the meantime, preserve the beneficial role of astroglial-fibrotic scar.Methods: In this study, we established an in vitro system, in which coculture of astrocytes and meningeal fibroblasts was treated with exogenous transforming growth factor-beta 1 (TGF-beta 1) to form astroglial-fibrotic scar-like cell clusters, and thereby evaluated the efficacy of RNAi targeting ephrin-B2 in preventing scar formation from the very beginning. We further tested the effect of RNAi-based mitigation of astroglial-fibrotic scar on spinal axon outgrowth on a custom-made microfluidic platform.Results: We found that siRNA targeting ephrin-B2 significantly reduced both the number and the diameter of cell clusters induced by TGF-beta 1 and diminished the expression of aggrecan and versican in the coculture, and allowed for significantly longer extension of outgrowing spinal cord axons into astroglial-fibrotic scar as assessed on the microfluidic platform.Conclusions: These results suggest that astroglial-fibrotic scar formation and particularly the expression of aggrecan and versican could be mitigated by ephrin-B2 specific siRNA, thus improving the microenvironment for spinal axon regeneration.