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牛乳铁蛋白通过细胞外信号调节激酶1/2保护RSC96雪旺细胞免受肿瘤坏死因子-α诱导的生长停滞。

Bovine lactoferrin protects RSC96 Schwann cells from tumor necrosis factor-alpha-induced growth arrest via extracellular-signal-regulated kinase 1/2.

作者信息

Lin T-Y, Kuo C-D

机构信息

Biophysics Laboratory, Department of Research and Education, Taipei Veterans General Hospital, Taipei 112, Taiwan.

出版信息

Neuroscience. 2008 Jan 24;151(2):396-402. doi: 10.1016/j.neuroscience.2007.10.018. Epub 2007 Oct 30.

Abstract

Bovine lactoferrin (bLF) is a member of the transferrin protein family that is abundant in bovine milk. Recent studies have shown that bLF plays roles in the regulation of cell growth. However, the biological function of bLF in the peripheral nervous system is still unclear. In this study, the immortalized rat Schwann cells (RSC96) were exposed to bLF at concentrations ranging from 10 to 800 microg/ml for 48 h and compared with control group. The bromodeoxyuridine BrdU cell viability assay was used to examine the effect of bLF on cell viability of RSC96 Schwann cells. Cell-counting test was used to assay the growth rate of RSC96 cells after exposure to bLF, and immunoblot analysis was used to test the signaling pathway controlled by bLF in the RSC96 cells. It was found that the viability of the RSC96 cells was increased by more than 25% when treated with 50 microg/ml bLF and the cell number of RSC96 cells was increased by more than threefold when treated with 800 microg/ml bLF. Our results showed that bLF could significantly improve viability and number of RSC96 Schwann cells. Also, bLF could significantly increase the phosphorylation state of extracellular-signal regulated kinase 1/2 (ERK1/2) that could be specifically inhibited by PD98059. Furthermore, bLF could not only protect RSC96 cells from tumor necrosis factor-alpha (TNF-alpha) -induced growth arrest but could also restore proliferation rate in TNF-alpha-treated RSC96 cells. In conclusion, bLF plays a crucial role in the proliferation of RSC96 Schwann cells and the protection of RSC96 Schwann cells from TNF-alpha-induced growth arrest via ERK1/2 protein.

摘要

牛乳铁蛋白(bLF)是转铁蛋白家族的成员,在牛乳中含量丰富。最近的研究表明,bLF在细胞生长调节中发挥作用。然而,bLF在周围神经系统中的生物学功能仍不清楚。在本研究中,将永生化大鼠雪旺细胞(RSC96)暴露于浓度范围为10至800微克/毫升的bLF中48小时,并与对照组进行比较。采用溴脱氧尿苷BrdU细胞活力测定法检测bLF对RSC96雪旺细胞活力的影响。细胞计数试验用于检测RSC96细胞暴露于bLF后的生长速率,免疫印迹分析用于检测bLF在RSC96细胞中控制的信号通路。结果发现,用50微克/毫升bLF处理时,RSC96细胞的活力增加了25%以上,用800微克/毫升bLF处理时,RSC96细胞的数量增加了三倍以上。我们的结果表明,bLF可以显著提高RSC96雪旺细胞的活力和数量。此外,bLF可以显著增加细胞外信号调节激酶1/2(ERK1/2)的磷酸化状态,而PD98059可以特异性抑制这种状态。此外,bLF不仅可以保护RSC96细胞免受肿瘤坏死因子-α(TNF-α)诱导的生长停滞,还可以恢复TNF-α处理的RSC96细胞的增殖速率。总之,bLF在RSC96雪旺细胞的增殖以及通过ERK1/2蛋白保护RSC96雪旺细胞免受TNF-α诱导的生长停滞中起着关键作用。

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