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Mechanical injury and IL-1β regulated LOXs and MMP-1, 2, 3 expression in ACL fibroblasts co-cultured with synoviocytes
Biotechnology Letters ( IF 2.0 ) Pub Date : 2020-05-01 , DOI: 10.1007/s10529-020-02870-9
Chunli Wang 1 , Qingjia Chi 1, 2 , Yongqiang Sha 1 , Kang Xu 1 , Chunming Xu 1 , Cheng Chen 3 , Wei Huang 3 , Peixing Chen 1 , Peter Chen 4 , Li Yang 1 , K L Paul Sung 1, 4
Affiliation  

Objective Interleukin (IL)-1β in the joint cavity increases to promote healing after anterior cruciate ligament (ACL) injury. Synovial tissue is a major joint microenvironmental regulator after ACL injury. The purpose of this study was to investigate the effects of synovial cells (SCs) on lysyl oxidase (LOX) and matrix metalloproteinase (MMP) production by ACL fibroblasts (ACLfs) in the presence of IL-1β. Results This study sheds light on the regulation of LOX and MMP-1, -2, -3 expression by ACLfs co-cultured with SCs and treated with IL-1β. LOX and MMP-1, 2, 3 gene/protein expression in IL-1β/stretch-stimulated ACLfs co-cultured with SCs were measured by real-time quantitative PCR and Western blot. Meanwhile, MMP-2 activity was analyzed by zymogram. The results showed that co-culture with SCs increased LOX and MMP-1, -2, -3 gene and protein expression in the presence of IL-1β. Next, ACLfs were subjected to 12% mechanical stretch to simulate pathological injury. Under these conditions, SCs inhibited IL-1β-mediated upregulation of LOXs. However, IL-1β enhanced the expression of MMP-1, -2, -3 in injured ACLfs. Conclusions SCs can either inhibit or increase LOX production in the presence of IL-1β, while promoting the accumulation of MMP in injured ACLfs. These results may provide crucial insights into the mechanisms underlying ACL poor healing capacity after injury.

中文翻译:

机械损伤和 IL-1β 调节与滑膜细胞共培养的 ACL 成纤维细胞中的 LOX 和 MMP-1、2、3 表达

目的前交叉韧带(ACL)损伤后,关节腔内白细胞介素(IL)-1β增加促进愈合。滑膜组织是 ACL 损伤后的主要关节微环境调节剂。本研究的目的是研究滑膜细胞 (SCs) 在 IL-1β 存在下对 ACL 成纤维细胞 (ACLfs) 产生赖氨酰氧化酶 (LOX) 和基质金属蛋白酶 (MMP) 的影响。结果 本研究阐明了与 SCs 共培养并用 IL-1β 处理的 ACLfs 对 LOX 和 MMP-1、-2、-3 表达的调节。通过实时定量 PCR 和蛋白质印迹法测量与 SCs 共培养的 IL-1β/拉伸刺激的 ACLfs 中的 LOX 和 MMP-1、2、3 基因/蛋白质表达。同时,通过酶谱分析MMP-2活性。结果表明,与 SCs 共培养增加了 LOX 和 MMP-1、-2、-3 在 IL-1β 存在下的基因和蛋白质表达。接下来,ACLfs 受到 12% 的机械拉伸以模拟病理损伤。在这些条件下,SCs 抑制了 IL-1β 介导的 LOX 上调。然而,IL-1β 增强了受损 ACLf 中 MMP-1、-2、-3 的表达。结论 SCs 可以在 IL-1β 存在的情况下抑制或增加 LOX 的产生,同时促进 MMP 在受伤的 ACLfs 中的积累。这些结果可能为 ACL 损伤后愈合能力差的潜在机制提供重要见解。结论 SCs 可以在 IL-1β 存在的情况下抑制或增加 LOX 的产生,同时促进 MMP 在受伤的 ACLfs 中的积累。这些结果可能为 ACL 损伤后愈合能力差的潜在机制提供重要见解。结论 SCs 可以在 IL-1β 存在的情况下抑制或增加 LOX 的产生,同时促进 MMP 在受伤的 ACLfs 中的积累。这些结果可能为 ACL 损伤后愈合能力差的潜在机制提供重要见解。
更新日期:2020-05-01
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