Institute of Clinical Chemistry, Hannover Medical School, 30625 Hannover, Germany.
Central Pharmacy, Hannover Medical School, 30625 Hannover, Germany.
Int J Mol Sci. 2019 Mar 30;20(7):1595. doi: 10.3390/ijms20071595.
Blood sampling with different anticoagulants alters matrix metalloproteinase (MMP-) 9 expression, thus influencing its concentration and diagnostic validity. Here, we aimed to evaluate the effects of different anticoagulants on MMP-9 regulation. MMP-9 expression was assessed in response to ethylenediaminetetraacetic acid, citrate, and high-/low-molecular-weight heparin (HMWH, LMWH) in co-culture experiments using THP-1, Jurkat, and HT cells (representing monocytes, T, and B cells). Triple and double cell line co-culture experiments revealed that HMWH treatment of THP-1 and Jurkat led to a significant MMP-9 induction, whereas other anticoagulants and cell type combinations had no effect. Supernatant of HMWH-treated Jurkat cells also induced MMP-9 in THP-1 suggesting monocytes as MMP-9 producers. HMWH-induced cytokine/chemokine secretion was assessed in co-culture supernatant, and the influence of cytokines/chemokines on MMP-9 production was analyzed. These experiments revealed that Jurkat-derived IL-16 and soluble intercellular adhesion molecule (sICAM-) 1 are able to induce MMP-9 and IL-8 production by THP-1. As a consequence, the increased MMP-9 expression found in HMWH blood samples may be influenced by HMWH-dependent secretion of IL-16 and sICAM-1 by T cells resulting in an increased production of MMP-9 and IL-8 by monocytes. IL-8, in turn, may support MMP-9 and its own expression in a positive autocrine feedback loop.
不同抗凝剂的血液采样会改变基质金属蛋白酶(MMP-9)的表达,从而影响其浓度和诊断的有效性。在这里,我们旨在评估不同抗凝剂对 MMP-9 调节的影响。采用 THP-1、Jurkat 和 HT 细胞(分别代表单核细胞、T 和 B 细胞)的共培养实验,评估 EDTA、柠檬酸盐和高/低分子量肝素(HMWH、LMWH)对 MMP-9 表达的影响。三重和双重细胞系共培养实验表明,HMWH 处理 THP-1 和 Jurkat 导致 MMP-9 的显著诱导,而其他抗凝剂和细胞类型组合则没有影响。HMWH 处理的 Jurkat 细胞的上清液也诱导 THP-1 中的 MMP-9,表明单核细胞是 MMP-9 的产生细胞。在共培养上清液中评估了 HMWH 诱导的细胞因子/趋化因子分泌,并分析了细胞因子/趋化因子对 MMP-9 产生的影响。这些实验表明,Jurkat 衍生的 IL-16 和可溶性细胞间黏附分子(sICAM-1)能够诱导 THP-1 产生 MMP-9 和 IL-8。因此,在 HMWH 血液样本中发现的 MMP-9 表达增加可能受到 T 细胞依赖 HMWH 分泌的 IL-16 和 sICAM-1 的影响,导致单核细胞产生更多的 MMP-9 和 IL-8。反过来,IL-8 可能在正反馈回路中支持 MMP-9 和其自身的表达。