Takahashi K, Kubo T, Kobayashi K, Imanishi J, Takigawa M, Arai Y, Hirasawa Y
Department of Orthopaedic Surgery, Kyoto Prefectural University of Medicine, Japan.
J Orthop Res. 1997 Jan;15(1):150-8. doi: 10.1002/jor.1100150122.
The present study investigated the influence of hydrostatic pressure on the expression of cytokines and heat shock protein 70 in a chondrocyte-like cell line. Chondrocyte-like cells (HCS-2/8) were exposed to hydrostatic pressure by a special pressure apparatus. Total RNA for cytokines (interleukin-1 beta, basic fibroblast growth factor, insulin-like growth factor-I, and transforming growth factor-beta 1) and for heat shock protein 70 was extracted and was analyzed by a polymerase chain reaction method and Northern blotting. An assay for incorporation of [35S]sulfate was performed to assess proteoglycan synthesis. The expression of transforming growth factor-beta 1 mRNA was enhanced after exposure to 5-MPa of hydrostatic pressure and was reduced after 50 MPa, whereas the expression of heat shock protein 70 was enhanced following exposure to 50 MPa of hydrostatic pressure. The incorporation of [35S]sulfate into the cultured cells increased following exposure to 1-5 MPa of hydrostatic pressure and decreased following 10-50 MPa of pressure. These results suggest that hydrostatic pressure at physiologic levels enhances the expression of transforming growth factor-beta 1 mRNA in addition to increasing proteoglycan synthesis in chondrocytes and that excessively high hydrostatic pressure reduces the expression of transforming growth factor-beta 1 mRNA and increases the expression of heat shock protein 70 mRNA while decreasing proteoglycan synthesis.
本研究调查了流体静压力对软骨样细胞系中细胞因子和热休克蛋白70表达的影响。通过特殊的压力装置使软骨样细胞(HCS-2/8)暴露于流体静压力下。提取细胞因子(白细胞介素-1β、碱性成纤维细胞生长因子、胰岛素样生长因子-I和转化生长因子-β1)和热休克蛋白70的总RNA,并通过聚合酶链反应法和Northern印迹法进行分析。进行[35S]硫酸盐掺入测定以评估蛋白聚糖的合成。暴露于5兆帕流体静压力后,转化生长因子-β1 mRNA的表达增强,而在50兆帕后降低,而暴露于50兆帕流体静压力后,热休克蛋白70的表达增强。暴露于1-5兆帕流体静压力后,[35S]硫酸盐掺入培养细胞增加,而在10-50兆帕压力后减少。这些结果表明,生理水平的流体静压力除了增加软骨细胞中蛋白聚糖的合成外,还增强了转化生长因子-β1 mRNA的表达,而过高的流体静压力会降低转化生长因子-β1 mRNA的表达并增加热休克蛋白70 mRNA的表达,同时减少蛋白聚糖的合成。