Institute of Clinical Chemistry and Laboratory Medicine, Hannover Medical School, 30625 Hannover, Germany.
Faculty of Pharmacy and Medical Sciences, Hebron University, Hebron 711, Palestine.
Int J Mol Sci. 2024 Sep 20;25(18):10106. doi: 10.3390/ijms251810106.
In contrast to other common anticoagulants such as citrate and low-molecular-weight heparin (LMWH), high-molecular-weight heparin (HMWH) induces the expression of matrix metalloproteinase (MMP)-9, which is also measured as a biomarker for stroke in blood samples. Mechanistically, HMWH-stimulated T cells produce cytokines that induce monocytic MMP-9 expression. Here, the influence of further anticoagulants (Fondaparinux, Hirudin, and Alteplase) and the heparin-contaminating glycosaminoglycans (GAG) hyaluronic acid (HA), dermatan sulfate (DS), chondroitin sulfate (CS), and over-sulfated CS (OSCS) on MMP-9 was analyzed to assess its suitability as a biomarker under various conditions. Therefore, starved Jurkat T cells were stimulated with anticoagulants/contaminants. Subsequently, starved monocytic THP-1 cells were incubated with the conditioned Jurkat supernatant, and MMP-9 mRNA levels were monitored (quantitative (q)PCR). Jurkat-derived mediators secreted in response to anticoagulants/contaminants were also assessed (proteome profiler array). The supernatants of HMWH-, Hirudin-, CS-, and OSCS-treated Jurkat cells comprised combinations of activating mediators and led to a significant (in the case of OSCS, dramatic) MMP-9 induction in THP-1. HA induced MMP-9 only in high concentrations, while LMWH, Fondaparinux, Alteplase, and DS had no effect. This indicates that depending on molecular weight and charge (but independent of anticoagulant activity), anticoagulants/contaminants provoke the expression of T-cell-derived cytokines/chemokines that induce monocytic MMP-9 expression, thus potentially impairing the diagnostic validity of MMP-9.
与其他常见的抗凝剂（如柠檬酸盐和低分子量肝素[LMWH]）不同，高分子量肝素（HMWH）会诱导基质金属蛋白酶（MMP）-9 的表达，MMP-9 也可作为血液样本中中风的生物标志物进行测量。从机制上讲，HMWH 刺激的 T 细胞产生细胞因子，诱导单核细胞 MMP-9 的表达。在这里，分析了其他抗凝剂（磺达肝素、水蛭素和阿替普酶）和肝素污染的糖胺聚糖（GAG）透明质酸（HA）、硫酸皮肤素（DS）、硫酸软骨素（CS）和过硫酸化 CS（OSCS）对 MMP-9 的影响，以评估其在各种条件下作为生物标志物的适用性。因此，饥饿的 Jurkat T 细胞用抗凝剂/污染物刺激。随后，饥饿的单核细胞 THP-1 细胞用条件 Jurkat 上清液孵育，并监测 MMP-9 mRNA 水平（定量（q）PCR）。还评估了 Jurkat 细胞对抗凝剂/污染物反应分泌的介质（蛋白质组分析器阵列）。HMWH、水蛭素、CS 和 OSCS 处理的 Jurkat 细胞的上清液包含激活介质的组合，导致 THP-1 中 MMP-9 的显著（在 OSCS 的情况下，明显）诱导。HA 仅在高浓度下诱导 MMP-9，而 LMWH、磺达肝素、阿替普酶和 DS 没有作用。这表明，取决于分子量和电荷（但与抗凝活性无关），抗凝剂/污染物会引发 T 细胞衍生的细胞因子/趋化因子的表达，从而诱导单核细胞 MMP-9 的表达，从而可能损害 MMP-9 的诊断有效性。
