Abstract

Lysophosphatidylcholine (LPC) is a major atherogenic lipid that stimulates an increase in mitochondrial reactive oxygen species (mtROS) and the release of cytokines under inflammasome activation. However, the potential receptors of LPC in macrophages are poorly understood. Members of the transient receptor potential (TRP) channel superfamily, which is crucially involved in transducing environmental irritant stimuli into nociceptor activity, are potential receptors of LPC. In this study, we investigated whether LPC can induce the activation of transient receptor potential ankyrin 1 (TRPA1), a member of the TRP superfamily. The functional expression of TRPA1 was first detected by quantitative real-time polymerase chain reaction (qRT-PCR), western blotting and calcium imaging in human acute monocytic leukemia cell line (THP-1)-derived macrophages. The mechanism by which LPC induces the activation of macrophages through TRPA1 was verified by cytoplasmic and mitochondrial calcium imaging, mtROS detection, a JC-1 assay, enzyme-linked immunosorbent assay, the CCK-8 assay and the lactate dehydrogenase (LDH) cytotoxic assay. LPC induced the activation of THP-1-derived macrophages via calcium influx, and this activation was suppressed by potent and selective inhibitors of TRPA1. These results indicated that TRPA1 can mediate mtROS generation, mitochondrial membrane depolarization, the secretion of IL-1β and cytotoxicity through cellular and mitochondrial Ca²⁺ influx in LPC-treated THP-1-derived macrophages. Therefore, the inhibition of TRPA1 may protect THP-1-derived macrophages against LPC-induced injury.

Graphic Abstract

中文翻译：

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瞬态受体电位锚蛋白1有助于溶血磷脂酰胆碱诱导的细胞内钙调节和THP-1衍生的巨噬细胞活化。

摘要

溶血磷脂酰胆碱（LPC）是主要的致动脉粥样硬化脂质，可刺激线粒体活性氧（mtROS）的增加以及在炎性体激活下释放细胞因子。但是，对LPC在巨噬细胞中的潜在受体了解甚少。LPC的潜在受体是瞬时受体电位（TRP）通道超家族的成员，该家族主要参与将环境刺激性刺激转化为伤害感受器活性。在这项研究中，我们调查了LPC是否可以诱导瞬时受体电位锚蛋白1（TRPA1）（TRP超家族的成员）的激活。首先通过实时定量聚合酶链反应（qRT-PCR），蛋白质印迹和钙成像在人急性单核细胞白血病细胞系（THP-1）衍生的巨噬细胞中检测TRPA1的功能表达。通过细胞质和线粒体钙成像，mtROS检测，JC-1分析，酶联免疫吸附分析，CCK-8分析和乳酸脱氢酶（LDH）细胞毒性分析验证了LPC通过TRPA1诱导巨噬细胞激活的机制。 。LPC通过钙内流诱导了THP-1衍生的巨噬细胞的活化，而该活化被有效和选择性的TRPA1抑制剂所抑制。这些结果表明TRPA1可通过细胞和线粒体钙介导mtROS的产生，线粒体膜去极化，IL-1β的分泌以及细胞毒性。CCK-8分析法和乳酸脱氢酶（LDH）细胞毒性分析法。LPC通过钙内流诱导了THP-1衍生的巨噬细胞的活化，而该活化被有效和选择性的TRPA1抑制剂所抑制。这些结果表明TRPA1可通过细胞和线粒体Ca介导mtROS的产生，线粒体膜去极化，IL-1β的分泌以及细胞毒性。CCK-8分析法和乳酸脱氢酶（LDH）细胞毒性分析法。LPC通过钙内流诱导了THP-1衍生的巨噬细胞的活化，而该活化被有效和选择性的TRPA1抑制剂所抑制。这些结果表明TRPA1可通过细胞和线粒体钙介导mtROS的产生，线粒体膜去极化，IL-1β的分泌以及细胞毒性。在LPC处理的THP-1衍生的巨噬细胞中有²⁺流入。因此，抑制TRPA1可以保护THP-1衍生的巨噬细胞免受LPC诱导的损伤。

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