State Key Laboratory of Oral Diseases, West China School of Stomatology, Sichuan University, Chengdu, 610041, China.
Cell Prolif. 2015 Feb;48(1):59-66. doi: 10.1111/cpr.12151. Epub 2014 Nov 24.
This study aimed to investigate the response of Schwann cells to cyclic compressive and tensile stresses of different durations of stimulation.
RSC96 cells were subjected to cyclic tensile stress or compressive stress; for either, cells in five groups were treated for 0, 1, 2, 24 and 48 h respectively. Enzyme-linked immunosorbent assay was conducted to detect secretion of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 and neurotrophin-4 in the culture medium. Real-time PCR was conducted to quantify mRNA expression of neurotrophins including NGF, BDNF, neurotrophin-3 and neurotrophin-4, and myelin-related genes including Sox10, Krox20, neuregulin 1, NCAM, N-cadherin, P0, MAG and MBP. Immunofluorescent staining was performed to visualize Krox20 and F-actin in the tensile groups.
Within 24 h, cells treated with cyclic tensile stress expressed and secreted significantly more BDNF, while cyclic compression down-regulated BDNF expression. Cells treated with both tensile and compressive stress down-regulated expression of NRG1, NCAM, Krox20 and Sox10 at all time points. Expression of N-cadherin was not affected by either stretch or compression. F-actin was down-regulated by tensile stress.
Both tensile and compressive loading down-regulated expression of several important myelin-related Schwann cells genes and thus facilitated demyelination. Tensile stress meanwhile promoted secretion of BDNF by Schwann cells within 24 h, which may contribute to maintenance and repair of damaged axons. These effects of mechanical stress might have been mediated by the actin cytoskeleton.
本研究旨在探讨雪旺细胞对不同刺激时长的循环压缩和拉伸应力的反应。
将 RSC96 细胞进行循环拉伸或压缩应变处理;对于任一种应变,将细胞分为五组,分别处理 0、1、2、24 和 48 h。采用酶联免疫吸附试验检测培养基中神经生长因子(NGF)、脑源性神经营养因子(BDNF)、神经营养因子-3 和神经营养因子-4 的分泌情况。采用实时 PCR 技术定量检测神经生长因子(NGF、BDNF、神经营养因子-3 和神经营养因子-4)和髓鞘相关基因(Sox10、Krox20、神经调节蛋白 1、NCAM、N-钙黏蛋白、P0、MAG 和 MBP)的 mRNA 表达。采用免疫荧光染色技术观察拉伸组中 Krox20 和 F-肌动蛋白的表达情况。
在 24 h 内,经循环拉伸处理的细胞表达和分泌的 BDNF 明显增多,而循环压缩则下调了 BDNF 的表达。经拉伸和压缩处理的细胞在所有时间点均下调了 NRG1、NCAM、Krox20 和 Sox10 的表达。N-钙黏蛋白的表达不受拉伸或压缩的影响。F-肌动蛋白受拉伸应力下调。
拉伸和压缩负荷均下调了几个重要的髓鞘相关雪旺细胞基因的表达,从而促进脱髓鞘。同时,拉伸应力在 24 h 内促进了雪旺细胞 BDNF 的分泌,这可能有助于受损轴突的维持和修复。这些机械应力的影响可能是通过肌动蛋白细胞骨架介导的。