Effects of Low-Intensity Pulsed Ultrasound on the Expression and Activity of Hyaluronan Synthase and Hyaluronidase in IL-1β-Stimulated Synovial Cells
Effects of Low-Intensity Pulsed Ultrasound on the Expression and Activity of Hyaluronan Synthase and Hyaluronidase in IL-1β-Stimulated Synovial Cells
复制标题
DOI：
10.1007/s10439-010-0104-5
复制
发表时间：
2010-11-01
影响因子：
3.8
通讯作者：
Tanaka, Eiji
中科院分区：
文献类型：
--
作者：
Nakamura, Tatsuya;Fujihara, Shinji;Tanaka, Eiji
本研究旨在探讨低强度脉冲超声(LIPUS)对IL-1β刺激的滑膜细胞透明质酸(HA)代谢的影响。将兔膝关节滑膜细胞系HIG-82加入或不加1 ng/mLIL-1β的培养液中培养，4h后加入LIPUS 15min。实时定量聚合酶链式反应(Real-Time-PCR)检测透明质酸合成酶(HAS)2、3、透明质酸酶(HYAL)2和环氧合酶(COX)-2的mRNA水平。用双抗体夹心酶联免疫吸附试验(ELISA)测定HA和PGE(2)的浓度。用免疫印迹法检测COX-2的表达。LIPUS可显著上调IL-1β刺激细胞中HAS2和HAS3的基因水平(p<；0.01)。经IL-1β处理后，Hyal2mRNA表达上调，而在接下来的LIPUS暴露中显著下调(p<；0.01)。IL-1β刺激可促进COX-2和PGE(2)的表达，而LIPUS可抑制IL-1β诱导的COX-2和PGE(2)表达。这些结果表明，LIPUS促进了HA的合成，抑制了Hyal2的表达，导致了高分子量HA的积累。因此，LIPUS刺激可能是治疗炎性关节疾病合并滑液中HA降解的较好药物。
The purpose of this study is to examine effects of low-intensity pulsed ultrasound (LIPUS) on metabolism of hyaluronan (HA) in synovial membrane cells stimulated by IL-1 beta. Rabbit knee synovial membrane cell line, HIG-82, was cultured in medium with the presence or absence of 1 ng/mL IL-1 beta, and after 4 h the cell was exposed to LIPUS for 15 min. The mRNA levels of HA synthase (HAS) 2,3, hyaluronidase (HYAL) 2, and cyclooxygenase (COX)-2 were examined by real-time PCR analysis. Concentrations of HA and PGE(2) were quantified by use of enzyme linked immunosorbent assay (ELISA). The COX-2 level was analyzed by western blotting. Gene levels of HAS2 and HAS3 in IL-1 beta-stimulated cells were up-regulated significantly (p < 0.01) by LIPUS. HYAL2 mRNA was up-regulated by the treatment with IL-1 beta, whereas down-regulated significantly (p < 0.01) by the following LIPUS exposure. Furthermore, IL-1 beta stimulation enhanced COX-2 and PGE(2) expression as compared to the untreated control, and IL-1 beta-induced COX-2 and PGE(2) expression was inhibited by LIPUS. These results suggest that LIPUS enhanced HA synthesis and inhibited HYAL2 expression, leading to the accumulation of high-molecular weight HA. Therefore, LIPUS stimulation may be a better candidate as medical remedy to treat inflammatory joint diseases accompanied with HA degradation in synovial fluid.