Laboratory for Experimental Orthopaedics, Department of Orthopaedics, Justus Liebig University Giessen, 35392 Giessen, Germany.
Mathematical Institute, Justus Liebig University Giessen, 35392 Giessen, Germany.
Int J Mol Sci. 2022 Feb 22;23(5):2409. doi: 10.3390/ijms23052409.
(1) Background: Synovial fluid (SF) from knee joints with osteoarthritis (OA) has increased levels of phospholipids (PL). We have reported earlier that TGF-ß and IGF-1 stimulate fibroblast-like synoviocytes (FLS) to synthesize increased amounts of PLs. The current study examined whether IL-1ß induces the release of PLs in FLS and the underlying mechanism. (2) Methods: Cultured human OA FLS were treated with IL-1ß alone and with pathway inhibitors or with synthetic liver X receptor (LXR) agonists. Cholesterol hydroxylases, ABC transporters, apolipoproteins (APO), LXR, sterol regulatory binding proteins (SREBPs), and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) were analyzed by RT-PCR, Western blot, and ELISA. The release of radiolabeled PLs from FLS was determined, and statistical analysis was performed using R (N = 5-9). (3) Results: Like synthetic LXR agonists, IL-1ß induced a 1.4-fold greater release of PLs from FLS. Simultaneously, IL-1ß upregulated the level of the PL transporter ABCA1 and of cholesterol hydroxylases CH25H and CYP7B1. IL-1ß and T0901317 stimulated the expression of SREBP1c, whereas only T0901317 enhanced SREBP2, HMGCR, APOE, LXRα, and ABCG1 additionally. (4) Conclusions: IL-1ß partially controls PL levels in OA-SF by affecting the release of PLs from FLS. Our data show that IL-1ß upregulates cholesterol hydroxylases and thus the formation of oxysterols, which, as natural agonists of LXR, increase the level of active ABCA1, in turn enhancing the release of PLs.
(1) 背景:骨关节炎(OA)膝关节滑液(SF)中磷脂(PL)水平升高。我们之前报道过 TGF-β和 IGF-1 刺激成纤维样滑膜细胞(FLS)合成更多的 PL。本研究探讨了 IL-1β是否诱导 FLS 释放 PL 及其潜在机制。
(2) 方法:用 IL-1β单独或用通路抑制剂或合成肝 X 受体(LXR)激动剂处理培养的人 OA FLS。用 RT-PCR、Western blot 和 ELISA 分析胆固醇羟化酶、ABC 转运体、载脂蛋白(APO)、LXR、固醇调节元件结合蛋白(SREBPs)和 3-羟基-3-甲基戊二酰辅酶 A 还原酶(HMGCR)。测定 FLS 释放放射性标记的 PL,并使用 R(N = 5-9)进行统计分析。
(3) 结果:与合成 LXR 激动剂一样,IL-1β诱导 FLS 释放 PL 的能力增加了 1.4 倍。同时,IL-1β上调了 PL 转运体 ABCA1 和胆固醇羟化酶 CH25H 和 CYP7B1 的水平。IL-1β和 T0901317 刺激 SREBP1c 的表达,而只有 T0901317 额外增强 SREBP2、HMGCR、APOE、LXRα和 ABCG1。
(4) 结论:IL-1β通过影响 FLS 释放 PL 来部分控制 OA-SF 中的 PL 水平。我们的数据表明,IL-1β上调胆固醇羟化酶,从而形成氧化固醇,作为 LXR 的天然激动剂,增加活性 ABCA1 的水平,进而增强 PL 的释放。