The phase changes of M1/M2 phenotype of microglia/macrophage following oxygen-induced retinopathy in mice
The phase changes of M1/M2 phenotype of microglia/macrophage following oxygen-induced retinopathy in mice
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小鼠氧诱导视网膜病变后小胶质细胞/巨噬细胞M1/M2表型的相变
DOI：
10.1007/s00011-020-01427-w
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发表时间：
2021-01-01
影响因子：
6.7
通讯作者：
Liang, Xiaoling
中科院分区：
文献类型：
--
作者：
Li, Jia;Yu, Shanshan;Liang, Xiaoling
目的小胶质细胞/巨噬细胞的激活参与了多种眼部疾病的研究。然而，M1/M2表型小胶质细胞/巨噬细胞在氧诱导视网膜病变（OIR）病理过程中的单独作用尚不清楚。本研究探讨M1/M2小胶质细胞/巨噬细胞在C57BL/6J小鼠OIR中的作用及调控机制。此外，我们还证实了小胶质细胞/巨噬细胞在OIR自然过程中M1/M2移位的时相，这为进一步的研究奠定了基础。材料和方法C57BL/6j幼崽从出生后7（P7）到出生后12 （P12）暴露于高氧环境中，然后恢复到常氧环境。然后对小鼠实施安乐死，并在一系列时间点摘取眼睛以作进一步研究。采用免疫荧光染色和实时定量聚合酶链反应（qPCR）检测M1/M2表型小胶质细胞/巨噬细胞活性。western blot检测NF-kappa b-STAT3信号通路和il -4- stat6 - ppar - γ信号通路活性。结果P12后建立OIR模型时，小胶质细胞/巨噬细胞被激活。M1小胶质细胞/巨噬细胞活化出现在中央和外周视网膜的新生血管（NV）丛中，从小鼠回到常氧环境的P12开始，在P17达到峰值。在此期间，NF-kappa b-STAT3信号通路被激活，导致M1表型小胶质细胞/巨噬细胞极化上调，同时肿瘤坏死因子- α (tnf - α)、白细胞介素-6 （IL-6）、IL-1 β等炎性细胞因子表达增强。因此，从P12开始观察到NV簇，直到P17，体积继续增加。然而，M2型小胶质细胞/巨噬细胞活性在P17开始的OIR后期接管。il -4- stat6 - ppar - γ信号活性从P17开始上调，并在P20达到峰值，诱导M2型小胶质细胞极化，从而抑制炎症细胞因子和NV丛的自发消退。结论小胶质细胞/巨噬细胞积极参与小鼠OIR的自然过程，两种表型发挥不同的功能。调节小胶质细胞/巨噬细胞向M2表型极化的治疗可能是一种新的有前途的方法，用于治疗眼部新生血管疾病，如早产儿视网膜病变（ROP）、湿性年龄相关性黄斑变性（wAMD）和糖尿病视网膜病变（DR）。
Objective Microglia/macrophage activation is previously reported to be involved in various ocular diseases. However, the separate role of M1/M2 phenotype microglia/macrophage in the pathological process of oxygen-induced retinopathy (OIR) remains unknown. In this research, we explored the role and regulatory mechanism of M1/M2 microglia/macrophage in OIR in C57BL/6J mice. Furthermore, we demonstrated the time phase of M1/M2 shifting of microglia/macrophage during the natural process of OIR, which is very essential for further investigations. Materials and methods C57BL/6j pups were exposed to hyperoxia environment from postnatal 7(P7) to P12 then returned to normoxia. The mice were then euthanized, and the eyes were harvested at a series of time points for further investigation. The M1/M2 phenotype microglia/macrophage activity was presented by immunofluorescent staining and real-time quantitative polymerase chain reaction (qPCR). The NF-kappa b-STAT3 signaling and IL-4-STAT6-PPAR-gamma signaling pathway activity was examined by western blot analysis. Results The microglia/macrophage were activated when the OIR model was set up after P12. The M1 microglia/macrophage activation was found in neovascularization (NV) tufts in both central and peripheral retina, which started from P12 when the mice were returned to normoxia environment and peaked at P17. During this period of time, the NF-kappa b-STAT3 signaling pathway was activated, resulting in the upregulated M1 phenotype microglia/macrophage polarization, along with the enhanced inflammatory cytokine expression including tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), and IL-1 beta. Consequently, the NV tufts were observed from P12 and the volume continued to increase until P17. However, the M2 phenotype microglia/macrophage activity took over during the late phase of OIR started from P17. The IL-4-STAT6-PPAR-gamma signaling activity was upregulated from P17 and peaked at P20, inducing M2 phenotype microglia polarization, which consequently led to the inhibition of inflammatory cytokines and spontaneous regression of NV tufts. Conclusions Microglia/macrophage participate actively in the natural process of OIR in mice, and two phenotypes exert different functions. Treatment modulating microglia/macrophage polarize toward M2 phenotype might be a novel and promising method for ocular neovascular diseases such as retinopathy of prematurity (ROP), wet age-related macular degeneration (wAMD), and diabetic retinopathy (DR).