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雪旺细胞与脑源性神经营养因子共培养移植治疗实验性自身免疫性神经炎。

Transplantation of Schwann cells co-cultured with brain-derived neurotrophic factor for the treatment of experimental autoimmune neuritis.

机构信息

Department of Neurology, the Second Affiliated Hospital of Harbin Medical University, Heilongjiang 150086, China.

出版信息

J Neuroimmunol. 2013 Oct 15;263(1-2):83-90. doi: 10.1016/j.jneuroim.2013.08.004. Epub 2013 Aug 15.

Abstract

The aim of this study was to investigate the effects of transplantation of Schwann cells (SCs) co-cultured with brain-derived neurotrophic factor (BDNF) for the treatment of experimental autoimmune neuritis (EAN). Primary SCs were co-cultured with various BDNF concentrations, and the optimum concentration was determined by cell proliferation, and NGF and FGF levels. A rat model of EAN was established by immunization with 400μg of P2 peptide dissolved in Freund's complete adjuvant. SCs were labeled with CFSE and injected into the cisterna magna 14days after immunization. We found proliferation of SCs, and NGF and FGF levels were highest at a BDNF concentration of 50ng/mL. Compared with EAN group, SCs+BDNF group showed the lower paralysis scores from day 34 to day 45, and in sciatic nerves showed a significant decrease in inflammatory cell infiltration (involved CD4-, CD8- and CD68-positive cells) at days 25 and 35, an alleviated demyelination at days 35 and 45, and an increase in S-100-positive cells and a decrease in NGF-positive cells at each time point (P<0.05). Compared with the EAN group, the SCs+BDNF group showed, in sciatic nerves, the mRNA level of NGF was significantly decreased but that of S-100 was increased at day 25, the mRNA level of CCL3 was also remarkably reduced at day 35, and the mRNA level of CD11a, CCL3 and NGF was reduced but that of S-100 was elevated at day 45 (P<0.05). There were no differences in results between the SCs group and EAN group. In the end, we draw the conclusions that the exogenous SCs injected through cisterna magna can migrate to the injured peripheral nerves, BDNF promotes the proliferation and secretory function of SCs in vitro, and BDNF-treated SCs in vivo can reduce paralysis, inflammation, and demyelination and improve the self-repair capability of body in EAN.

摘要

本研究旨在探讨共培养脑源性神经营养因子(BDNF)的施万细胞(SCs)移植治疗实验性自身免疫性神经炎(EAN)的作用。原代SCs 与不同浓度的 BDNF 共培养,通过细胞增殖、NGF 和 FGF 水平确定最佳浓度。通过用 400μg 溶解在完全弗氏佐剂中的 P2 肽免疫建立 EAN 大鼠模型。免疫后 14 天,SCs 用 CFSE 标记并注入枕骨大孔。我们发现,SCs 的增殖以及 NGF 和 FGF 水平在 BDNF 浓度为 50ng/mL 时最高。与 EAN 组相比,SCs+BDNF 组从第 34 天到第 45 天的瘫痪评分较低,坐骨神经中的炎症细胞浸润(涉及 CD4-、CD8-和 CD68 阳性细胞)在第 25 天和第 35 天显著减少,脱髓鞘在第 35 天和第 45 天减轻,S-100 阳性细胞增加,NGF 阳性细胞减少在每个时间点(P<0.05)。与 EAN 组相比,SCs+BDNF 组在坐骨神经中,NGF 的 mRNA 水平在第 25 天显著降低,但 S-100 的 mRNA 水平升高,CCL3 的 mRNA 水平在第 35 天也明显降低,CD11a、CCL3 和 NGF 的 mRNA 水平降低,但 S-100 的 mRNA 水平在第 45 天升高(P<0.05)。SCs 组和 EAN 组的结果没有差异。最后,我们得出结论,通过枕骨大孔注射的外源性SCs 可以迁移到受损的周围神经,BDNF 促进 SCs 的体外增殖和分泌功能,体内 BDNF 处理的 SCs 可以减轻 EAN 中的瘫痪、炎症和脱髓鞘,并提高机体的自我修复能力。

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