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缺氧/复氧对原代培养雪旺细胞活力及神经营养因子(NTFs)表达和分泌的影响。

Effect of hypoxia/reoxygenation on cell viability and expression and secretion of neurotrophic factors (NTFs) in primary cultured schwann cells.

机构信息

Department of Anatomy, Shanghai Jiao Tong University School of Medicine, Shanghai Jiao Tong University Institutes of Medical Sciences, Shanghai, China.

出版信息

Anat Rec (Hoboken). 2010 May;293(5):865-70. doi: 10.1002/ar.21105.

Abstract

As the primary myelin-forming cells of the peripheral nervous system, Schwann cells (SCs) play a key role in the regeneration of injured peripheral nerves. However, hypoxia causes injury of SCs, as observed in peripheral neuropathies, including those caused by diabetes. So we investigated the effect of hypoxia/reoxygenation (H/R) on SCs in this study. To do so, SCs were cultured in hypoxic condition in vitro and then in normal condition for 24 hr; The effects H/R on SCs were evaluated by MTT (3(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide) assay, Hoechst staining, immunocytochemistry, western blotting, ELISA, and RT-PCR. H/R resulted in a significant decrease in SCs survival and an increase in caspase-3 activity. H/R also reduced the mRNA level of BDNF (brain derived neurotrophic factor) and its secretion, but NGF mRNA level was elevated in these cells. These observations showed that H/R induces death of primary cultured SCs, and different mechanisms responsible for regulating NGF and BDNF expression.

摘要

作为周围神经系统的主要髓鞘形成细胞,施万细胞(Schwann cells,SCs)在受伤周围神经的再生中起着关键作用。然而,缺氧会导致 SC 损伤,如在周围神经病变中观察到的那样,包括糖尿病引起的周围神经病变。因此,我们在这项研究中研究了缺氧/复氧(Hypoxia/Reoxygenation,H/R)对 SC 的影响。为此,将 SC 在体外缺氧条件下培养,然后在正常条件下培养 24 小时;通过 MTT(3(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)测定、Hoechst 染色、免疫细胞化学、Western blot、ELISA 和 RT-PCR 评估 H/R 对 SC 的影响。H/R 导致 SC 存活率显著降低,半胱天冬酶-3 活性增加。H/R 还降低了 BDNF(脑源性神经营养因子)的 mRNA 水平及其分泌,但这些细胞中的 NGF mRNA 水平升高。这些观察结果表明,H/R 诱导原代培养的 SC 死亡,并负责调节 NGF 和 BDNF 表达的不同机制。

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