Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
Ann Biomed Eng. 2010 Apr;38(4):1618-25. doi: 10.1007/s10439-010-9927-3. Epub 2010 Jan 23.
Hyaluronan (HA) plays a crucial role in the lubricating and buffering properties of synovial fluid. The purpose of this study was to examine the effects of interleukin (IL)-1beta on HA degradation in cultured synovial membrane cells. The rabbit synovial membrane cell line HIG-82 was cultured with and without IL-1beta. The amounts of HA of varying molecular weights in the medium were analyzed using high-performance liquid chromatography, the mRNA levels of HA synthase (HAS) and hyaluronidase (HYAL) were analyzed by means of real-time PCR, and HYAL activity was analyzed by HA zymography. The amounts of HA with a molecular weight lower than 300 kDa, and between 300 and 1900 kDa, in the culture medium of HIG-82 cells were significantly higher in the presence of IL-1beta. However, the amount of HA with a molecular weight greater than 1900 kDa was significantly lower in the presence of IL-1beta. Both HAS2 and HAS3 mRNA levels were upregulated by treatment with IL-1beta. So, too, were the levels of HYAL1 and HYAL2 mRNA, which resulted in enhanced HYAL activity. However, HYAL activity was inhibited by transfection of HYAL2-siRNA. Our results suggest that IL-1beta is a crucial factor in the fragmentation of HA in inflammatory joints.
透明质酸 (HA) 在滑液的润滑和缓冲特性中起着至关重要的作用。本研究旨在研究白细胞介素 (IL)-1β对培养的滑膜细胞膜细胞中 HA 降解的影响。用和不用白细胞介素 (IL)-1β培养兔滑膜膜细胞系 HIG-82。用高效液相色谱法分析培养基中不同分子量 HA 的含量，实时 PCR 分析 HA 合酶 (HAS) 和透明质酸酶 (HYAL) 的 mRNA 水平，HA 酶谱分析 HYAL 活性。在存在白细胞介素 (IL)-1β的情况下，HIG-82 细胞培养基中分子量低于 300 kDa 和 300 至 1900 kDa 的 HA 含量明显增加。然而，在存在白细胞介素 (IL)-1β的情况下，分子量大于 1900 kDa 的 HA 含量明显降低。白细胞介素 (IL)-1β处理后，HAS2 和 HAS3 的 mRNA 水平均上调。HYAL1 和 HYAL2 的 mRNA 水平也上调，导致 HYAL 活性增强。然而，HYAL2-siRNA 的转染抑制了 HYAL 活性。我们的结果表明，白细胞介素 (IL)-1β是炎症关节中 HA 片段化的关键因素。
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