Free heme is a danger signal inducing expression of proinflammatory proteins in cultured cells derived from normal rat hearts.
Free heme is a danger signal inducing expression of proinflammatory proteins in cultured cells derived from normal rat hearts.
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DOI：
10.1016/j.molimm.2011.02.013
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发表时间：
2011-05
影响因子：
3.6
通讯作者：
Kazuhisa Hao;H. Hanawa;Limin Ding;Y. Ota;Kaori Yoshida;K. Toba;M. Ogura;Hiromi Ito;M. Kodama;Y. Aizawa
中科院分区：
文献类型：
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作者：
Kazuhisa Hao;H. Hanawa;Limin Ding;Y. Ota;Kaori Yoshida;K. Toba;M. Ogura;Hiromi Ito;M. Kodama;Y. Aizawa
来自受损组织的内源性分子充当危险信号，触发或放大免疫/炎症反应。在这项研究中，我们检查了游离血红素是否在正常心脏的培养细胞中诱导促炎蛋白，并研究了血红素靶向的细胞及其在这些细胞中的作用机制。我们培养了胶原酶分离的正常大鼠心脏来源的细胞，并通过定量 RT-PCR、ELISA 和流式细胞术检查游离血红素是否诱导促炎蛋白、活性氧 (ROS) 产生和 NF-κB 激活。游离血红素增加培养的心脏驻留细胞 (CCRC) 中各种促炎蛋白的 mRNA（至少 100 倍）并诱导细胞内 ROS 形成。大约 85-90% 的 CCRC 是成纤维细胞/平滑肌细胞，10-15% 是 CD11bc 阳性巨噬细胞；因此，为了检查单个靶细胞，对巨噬细胞缺失（CD11bc阴性）CCRC、原代培养细胞（心脏成纤维细胞、动脉平滑肌细胞和心脏微血管内皮细胞）和巨噬细胞细胞系（NR8383）进行类似处理。游离血红素激活 NF-κB 并诱导一些促炎蛋白的表达，包括 NR8383 中的 IL-1 和 TNF-α。另一方面，巨噬细胞缺失的 CCRC 在用 IL-1 或 TNF-α 处理后，这些蛋白的表达显着增加，但游离血红素却没有增加。游离血红素诱导的促炎蛋白表达不受细胞内 ROS 减少的抑制，而是受到能够调节 NF-κB 的蛋白酶和蛋白酶体抑制剂的抑制。这些数据表明，游离血红素通过心脏驻留巨噬细胞中的 NF-κB 激活以及通过尤其由 IL-1、TNF-α 介导的巨噬细胞和成纤维细胞/平滑肌细胞之间的串扰强烈诱导受伤心脏中的各种促炎蛋白。
Endogenous molecules from damaged tissue act as danger signals to trigger or amplify the immune/inflammatory response. In this study, we examined whether free heme induced pro-inflammatory proteins in cultured cells derived from normal hearts and investigated the cells targeted by heme, together with its mechanism of action in these cells. We cultured collagenase-isolated heart-derived cells from normal rats and examined whether free heme induced pro-inflammatory proteins, reactive oxygen species (ROS) production and NF-κB activation, by quantitative RT-PCR, ELISA and flow cytometry. Free heme increased mRNA of various pro-inflammatory proteins in cultured cardiac resident cells (CCRC) (at least 100-fold) and induced intracellular ROS formation. Approximately 85–90% of CCRC are fibroblast/smooth muscle cells and 10–15% are CD11bc-positive macrophages; therefore to examine individual target cells, macrophage-deleted (CD11bc-negative) CCRC, primary cultured cells (cardiac fibroblasts, arterial smooth muscle cells and cardiac microvascular endothelial cells) and macrophage cells lines (NR8383) were similarly treated. Free heme activated NF-κB and induced expression of some pro-inflammatory proteins, including IL-1 and TNF-α in NR8383. On the other hand, macrophage-deleted CCRC strongly increased expression of these proteins on treatment with IL-1 or TNF-α, but not free heme. Induction of expression of pro-inflammatory proteins by free heme was not inhibited by intracellular ROS reduction, but by protease and proteasome inhibitors capable of regulating NF-κB. These data suggest that free heme strongly induces various pro-inflammatory proteins in injured hearts through NF-κB activation in cardiac resident macrophages and through cross-talk between macrophages and fibroblast/smooth muscle cells mediated inter alia by IL-1, TNF-α.