缺氧成像探针“F-FMISO”在巨噬细胞中的积累除了依赖于缺氧状态外，还依赖于巨噬细胞的极化。

Accumulation of hypoxia imaging probe "F-FMISO" in macrophages depends on macrophage polarization in addition to hypoxic state.

机构信息

Laboratory of Bioanalysis and Molecular Imaging, Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita 12, Nishi 6, Kita-ku, Sapporo, 060-0812, Japan.

Kyoto University Hospital, 54 Shogoinkawahara-cho, Sakyo-ku, Kyoto, 606-8507, Japan.

出版信息

Ann Nucl Med. 2019 May;33(5):362-367. doi: 10.1007/s12149-019-01332-1. Epub 2019 Jan 29.

DOI:10.1007/s12149-019-01332-1

PMID:30694443

Abstract

OBJECTIVE

Macrophages play an essential role in immune response, and are closely related to the progression of diseases such as cancer and atherosclerosis. Macrophages polarize to M1 or M2 type, which is related to the environmental hypoxic state. Previously, we found that F-FMISO uptake varied according to expression levels of biomolecules such as glutathione S-transferase P1 (GST-P1), which catalyzes the conjugation of glutathione to F-FMISO metabolites, and multidrug resistance-associated protein 1 (MRP1), which exports glutathione-F-FMISO metabolite conjugates out of cells. However, the relationship between macrophage polarization and F-FMISO accumulation remains unclear.

METHODS

Mouse peritoneal macrophages were polarized to either the M1 or M2 type, and were treated with F-FMISO. Then, their radioactivity after a 4 h incubation period under normoxic (21% O) or hypoxic (1% O) condition was measured. GST-P1 and MRP1 expression levels were measured by qRT-PCR.

RESULTS

M2 macrophages exhibited a significantly higher uptake of F-FMISO than non-polarized (M0) macrophages, whereas M1 macrophages had a significantly lower uptake than M0 macrophages (M0: 1.05 ± 0.22, M1: 0.34 ± 0.02, M2: 4.17 ± 0.36 %dose/mg protein). The GST-P1 expression level in M1 macrophages was higher than that in M2 and M0 macrophages [GST-P1/β-actin normalized by M0: 9.0 ± 3.7 (M1), 1.2 ± 0.2 (M2)]. The MRP1 expression level in M1 macrophages was significantly higher than that in M2 and M0 macrophages [MRP1/β-actin normalized by M0 macrophages: 5.1 ± 2.1 (M1), 2.8 ± 1.0 (M2)].

CONCLUSIONS

F-FMISO accumulation in macrophages may depend on the polarization state in addition to hypoxic condition.

摘要

目的

巨噬细胞在免疫反应中发挥重要作用，与癌症和动脉粥样硬化等疾病的进展密切相关。巨噬细胞向 M1 或 M2 型极化，这与环境缺氧状态有关。先前，我们发现 F-FMISO 的摄取量根据谷胱甘肽 S-转移酶 P1（GST-P1）等生物分子的表达水平而变化，GST-P1 催化谷胱甘肽与 F-FMISO 代谢物的结合，多药耐药相关蛋白 1（MRP1）将谷胱甘肽-F-FMISO 代谢物从细胞内输出。然而，巨噬细胞极化与 F-FMISO 积累之间的关系尚不清楚。

方法

将小鼠腹腔巨噬细胞极化为 M1 或 M2 型，并给予 F-FMISO。然后，在常氧（21% O）或缺氧（1% O）条件下孵育 4 小时后测量其放射性。通过 qRT-PCR 测量 GST-P1 和 MRP1 的表达水平。

结果

M2 巨噬细胞对 F-FMISO 的摄取明显高于非极化（M0）巨噬细胞，而 M1 巨噬细胞的摄取明显低于 M0 巨噬细胞（M0：1.05±0.22，M1：0.34±0.02，M2：4.17±0.36%剂量/毫克蛋白）。M1 巨噬细胞中的 GST-P1 表达水平高于 M2 和 M0 巨噬细胞[GST-P1/β-肌动蛋白标准化值为 M0：9.0±3.7（M1），1.2±0.2（M2）]。M1 巨噬细胞中的 MRP1 表达水平明显高于 M2 和 M0 巨噬细胞[M0 巨噬细胞标准化的 MRP1/β-肌动蛋白：5.1±2.1（M1），2.8±1.0（M2）]。