Pam3CSK4 Induces MMP-9 Expression in Human Monocytic THP-1 Cells
Pam3CSK4 Induces MMP-9 Expression in Human Monocytic THP-1 Cells
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DOI：
10.1159/000475298
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Ahmad, Rasheed
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文献类型：
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作者：
Al-Rashed, Fatema;Kochumon, Shihab;Ahmad, Rasheed
背景：基质金属蛋白酶-9可降解细胞外基质，基质金属蛋白酶-9水平升高与包括肥胖在内的许多炎症性疾病的发生有关。Pam3CSK4是一种人工合成的三酰化脂肽(LP)，是一种有效的免疫细胞激活剂，并能诱导细胞因子的产生。然而，Pam3CSK4是否能够诱导单核细胞表达基质金属蛋白酶-9尚不清楚。因此，我们测定了Pam3CSK4处理的THP-1细胞产生的基质金属蛋白酶-9，并研究了相关的信号转导途径(S)。方法：采用实时荧光定量聚合酶链式反应和酶联免疫吸附试验检测基质金属蛋白酶-9的表达。酶谱法检测基质金属蛋白酶-9活性。用THP-1细胞、THP1-XBlue(TM)细胞、THP1-XBlue(TM)-DefMyD细胞、抗TLR2单抗和选择性药物抑制剂研究相关的信号转导途径。Western blotting检测磷酸化蛋白和总蛋白。结果：Pam3CSK4可诱导人单核细胞THP-1细胞表达MMP9(P<0.05)。在Pam3CSK4处理的THP-1细胞中，可检测到NF-kappa B/AP-1的活性增加，而抗TLR2中和抗体或抑制依赖于笼蛋白的内吞作用可显著抑制这些细胞的基质金属蛋白酶-9的产生。经Pam3CSK4处理后，MyD88-/-THP-1细胞不表达基质金属蛋白酶-9。抑制JNK、MEK/ERK、p38MAPK和NF-kappaB可显著抑制MMP9基因的表达(P<0.05)。结论：Pam3CSK4通过TLR-2/MyD88依赖的MEK/ERK、JNK、p38MAPK和NF-kappa B/AP-1激活机制诱导THP-1细胞产生MMP9。(三)2017年提交人(S)。由S.Karger AG出版，巴塞尔。
Background: Matrix metalloproteinase (MMP)-9 is known to degrade the extracellular matrix and increased MMP-9 levels are related with the pathogenesis of many inflammatory conditions including obesity. Pam3CSK4 is a synthetic triacylated lipopeptide (LP) which is a potent activator of immune cells and induces cytokine production. However, it is unclear whether Pam3CSK4 is able to induce MMP-9 expression in monocytic cells. We, therefore, determined MMP-9 production by Pam3CSK4-treated THP-1 cells and also investigated the signal transduction pathway(s) involved. Methods: MMP-9 expression was determined by real-time qPCR and ELISA. MMP-9 activity was assessed by zymography. THP-1 cells, THP1-XBlue (TM) cells, THP1-XBlue (TM)-defMyD cells, anti-TLR2 mAb and selective pharmacological inhibitors were used to study signaling pathways involved. Phosphorylated and total proteins were detected by western blotting. Results: Pam3CSK4 induced MMP-9 expression (P < 0.05) at both mRNA and protein levels in human monocytic THP-1 cells. Increased NF-kappa B/AP-1 activity was detected in Pam3CSK4-treated THP-1 cells and MMP-9 production in these cells was significantly suppressed by pre-treatment with anti-TLR2 neutralizing antibody or by inhibition of clathrin-dependent endocytosis. Also, MyD88- /- THP-1 cells did not express MMP-9 following treatment with Pam3CSK4. Inhibition of JNK, MEK/ERK, p38 MAPK and NF-kappa B significantly suppressed MMP-9 gene expression (P < 0.05). Conclusion: Pam3CSK4 induces MMP-9 production in THP-1 cells through the TLR-2/MyD88-dependent mechanism involving MEK/ERK, JNK, p38 MAPK and NF-kappa B/AP-1 activation. (C) 2017 The Author(s). Published by S. Karger AG, Basel.