Macrophages Protect Endometriotic Cells Against Oxidative Damage Through a Cross-Talk Mechanism
Macrophages Protect Endometriotic Cells Against Oxidative Damage Through a Cross-Talk Mechanism
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DOI：
10.1007/s43032-022-00890-6
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发表时间：
2022-02-23
影响因子：
2.9
通讯作者：
Kobayashi，Hiroshi
中科院分区：
文献类型：
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作者：
Ogawa，Kenji;Liu，Tingting;Kobayashi，Hiroshi
本研究的目的是研究巨噬细胞是否保护增生细胞免受氧化损伤，并阐明任何保护的潜在机制。用过氧化氢（H2 O2）或高铁血红蛋白（高铁血红蛋白是子宫内膜异位囊肿液中血红蛋白的主要成分）处理有或无分化巨噬细胞（dTHP-1细胞）培养的子宫内膜异位细胞。共培养实验、微阵列分析、差异表达基因（DEG）的筛选和验证、细胞增殖和活力测定以及使用特异性抑制剂的实验进行以研究增生细胞和巨噬细胞之间的功能性串扰。微阵列分析显示，与dTHP-1共培养的分化细胞与单培养相比差异表达几个基因。定量酶联免疫吸附试验（ELISA）和Western blotting分析证实TGF-β1在与dTHP-1细胞共培养的人卵巢癌细胞中表达。TGF-β1刺激dTHP-1细胞血红素氧合酶-1（HO-1）的表达。与dTHP-1单独培养相比，dTHP-1细胞与凋亡细胞共培养后HO-1表达增加。H2 O2和高铁血红蛋白均能上调dTHP-1单培养物中HO-1蛋白的表达;此外，与细胞共培养进一步提高HO-1的产量。与dTHP-1的共培养保护增生细胞免受氧化损伤。阻断HO-1可使巨噬细胞的保护作用消失。在氧化应激环境中，由肥大细胞产生的TGF-β1可通过上调巨噬细胞源性HO-1来保护免受氧化损伤。异位细胞与巨噬细胞间的相互作用可能参与了子宫内膜异位症的发生发展。
This aim of this study was to investigate whether macrophages protect endometriotic cells from oxidative injury and to elucidate the underlying mechanisms of any protection. Endometriotic cells cultured with or without differentiated macrophages (dTHP-1 cells) were treated with hydrogen peroxide (H2O2) or methemoglobin, a major component of hemoglobin species in endometriotic cyst fluid. Co-culture experiments, microarray analysis, screening and validation of differentially expressed genes (DEGs), cell proliferation and viability assays, and experiments using a specific inhibitor were conducted to investigate the functional cross-talk between endometriotic cells and macrophages. Microarray analysis revealed that endometriotic cells co-cultured with dTHP-1 differentially express several genes compared with monoculture. Quantitative enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis identified TGF-β1 as a promising candidate gene expressed in endometriotic cells co-cultured with dTHP-1 cells. TGF-β1 stimulated the expression of heme oxygenase-1 (HO-1) in dTHP-1 cells. HO-1 expression was increased in dTHP-1 cells co-cultured with endometriotic cells compared with the dTHP-1 monoculture. Both H2O2and methemoglobin upregulated the expression of the HO-1 protein in the dTHP-1 monoculture; moreover, co-culture with endometriotic cells further enhanced HO-1 production. The co-culture with dTHP-1 protected endometriotic cells against oxidative injury. Blockade of HO-1 abolished the protective effects of macrophages. In an oxidative stress environment, TGF-β1 produced by endometriotic cells may protect against oxidative injury through the upregulation of macrophage-derived HO-1. The cross-talk between endometriotic cells and macrophages may contribute to the progression and pathogenesis of endometriosis.