Iron overloaded polarizes macrophage to proinflammation phenotype through ROS/acetyl-p53 pathway.
Iron overloaded polarizes macrophage to proinflammation phenotype through ROS/acetyl-p53 pathway.
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铁超载通过 ROS/乙酰基-p53 途径将巨噬细胞极化为促炎症表型
DOI：
10.1002/cam4.1670
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发表时间：
2018-08
期刊：
影响因子：
4
通讯作者：
Liu ZJ
中科院分区：
文献类型：
--
作者：
Zhou Y;Que KT;Zhang Z;Yi ZJ;Zhao PX;You Y;Gong JP;Liu ZJ
巨噬细胞在炎症和伤口愈合中起关键作用，可分为两种亚型：经典激活型（M1）和替代激活型（M2）巨噬细胞。巨噬细胞在调节铁稳态方面也起着重要作用，细胞内铁积累诱导M1型巨噬细胞极化，这为通过M2肿瘤相关巨噬细胞再极化进行肿瘤免疫治疗提供了一种潜在方法。然而，铁诱导M1极化的潜在机制仍不清楚。
采用蛋白质印迹法、实时定量聚合酶链反应（qRT - PCR）和流式细胞术检测铁处理后的RAW 264.7小鼠巨噬细胞的极化指标，并采用蛋白质印迹法和qRT - PCR检测p21的表达。采用2,7 - 二氯二氢荧光素二乙酸酯测量铁或N - 乙酰 - L - 半胱氨酸（NAC）处理后巨噬细胞中的活性氧（ROS）水平。采用p300/CREB结合蛋白（CBP）抑制剂C646抑制p53乙酰化，并采用蛋白质印迹法、qRT - PCR和免疫荧光检测p53的表达及乙酰化。将BALB/c小鼠皮下注射H22肝癌细胞，并在尾静脉注射铁后研究巨噬细胞极化状态。采用免疫组织化学染色评估皮下肿瘤中分化群86（CD86）和含类表皮生长因子模块的粘蛋白样激素受体样1（F4/80）的蛋白表达。
铁过载通过增加RAW细胞中ROS的产生和诱导p53乙酰化来诱导M1极化，而NAC降低ROS水平可抑制M1极化和p53乙酰化。p300/CBP抑制剂对乙酰化p53的抑制可阻止M1极化并抑制p21的表达。这些结果表明，铁过载诱导的高ROS水平通过增强p300/CBP乙酰转移酶活性和促进p53乙酰化使巨噬细胞极化为M1亚型。
Macrophages play critical roles in inflammation and wound healing and can be divided into two subtypes: classically activated (M1) and alternatively activated (M2) macrophages. Macrophages also play important roles in regulating iron homeostasis, and intracellular iron accumulation induces M1‐type macrophage polarization which provides a potential approach to tumor immunotherapy through M2 tumor‐associated macrophage repolarization. However, the mechanisms underlying iron‐induced M1 polarization remain unclear. Western blotting, qRT‐PCR, and flow cytometry were used to detect the polarization indexes in RAW 264.7 murine macrophages treated with iron, and Western bloting and qRT‐PCR were used to detect p21 expression. The compound 2,7‐dichlorofluorescein diacetate was used to measure reactive oxygen species (ROS) levels in macrophages after iron or N‐acetyl‐l‐cysteine (NAC) treatment. The p300/CREB‐binding protein (CBP) inhibitor C646 was used to inhibit p53 acetylation, and Western bloting, qRT‐PCR, and immunofluorescence were used to detect p53 expression and acetylation. BALB/c mice were subcutaneously injected with H22 hepatoma cells, and macrophage polarization status was investigated after tail intravenous injection of iron. Immunohistochemical staining was used to evaluate the protein expression of cluster of differentiation 86 (CD86) and EGF‐like module‐containing mucin‐like hormone receptor‐like 1 (F4/80) in the subcutaneous tumors. Iron overload induced M1 polarization by increasing ROS production and inducing p53 acetylation in RAW cells, and reduction in ROS levels by NAC repressed M1 polarization and p53 acetylation. Inhibition of acetyl‐p53 by a p300/CBP inhibitor prevented M1 polarization and inhibited p21 expression. These results showed that high ROS levels induced by iron overload polarized macrophages to the M1 subtype by enhancing p300/CBP acetyltransferase activity and promoting p53 acetylation.
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DOI：
10.1038/nri3423
发表时间：
2013-05
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
通讯作者：
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影响因子：
38.3
作者：
通讯作者：
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影响因子：
4.6
作者：
Funauchi Y;Tanikawa C;Yi Lo PH;Mori J;Daigo Y;Takano A;Miyagi Y;Okawa A;Nakamura Y;Matsuda K
通讯作者：
Matsuda K
影响因子：
20.3
作者：
Kuiper, Jan W. P.;Sun, Chunxiang;Glogauer, Michael
通讯作者：
Glogauer, Michael
影响因子：
3.3
作者：
Gaharwar, Usha Singh;Meena, Ramovatar;Rajamani, Paulraj
通讯作者：
Rajamani, Paulraj