Old age promotes retinal fibrosis in choroidal neovascularization through circulating fibrocytes and profibrotic macrophages.
Old age promotes retinal fibrosis in choroidal neovascularization through circulating fibrocytes and profibrotic macrophages.
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DOI：
10.1186/s12974-023-02731-y
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发表时间：
2023-02-23
影响因子：
9.3
通讯作者：
中科院分区：
文献类型：
--
作者：
视网膜纤维化影响40-70%的新生血管性年龄相关性黄斑变性患者。本研究探讨了年龄对继发于脉络膜新生血管的视网膜下纤维化的影响及其作用机制。使用两阶段激光方案在年轻（2.5个月）和老年（15-16个月）C57 BL/6 J小鼠中诱导视网膜下纤维化。5天和30天后，收集眼睛并进行CD 45和胶原-1染色，并通过共聚焦显微镜观察。通过流式细胞术和共聚焦显微镜分别检测骨髓（BM）、血液和纤维化病变中的纤维细胞（CD 45+胶原-1+）。在有或没有TGF-β1（10 ng/mL）处理的情况下，从年轻和老年小鼠培养BM衍生的巨噬细胞（BMDM）。通过qPCR和免疫细胞化学检查间充质标志物αSMA（Acta 2）、纤连蛋白（Fn 1）和胶原蛋白-1（Col 1a 1）的表达，而使用Luminex多重细胞因子测定法测量细胞因子/趋化因子的产生。将BM从22月龄（Ly5.2）小鼠移植到2.5月龄（Ly5.1）年轻小鼠中，反之亦然。六周后，在受体小鼠中诱导视网膜下纤维化，并收集眼睛用于评估纤维化病变大小。在正常条件下，循环纤维细胞（CD 45+胶原-1+）的数量和Tgfb 1，Col 1a 1，Acta 2和Fn 1在BMDM中的表达水平在老年小鼠中显著高于年轻小鼠。视网膜下纤维化的诱导显著增加了循环纤维细胞的数量，增加了Col 1a 1，Acta 2和Fn 1的表达和可溶性尿激酶纤溶酶原激活物表面受体（suPAR）的产生，但减少了BMDM中CXCL 10的产生。来自老年视网膜下纤维化小鼠的BMDM比来自年轻视网膜下纤维化小鼠的细胞产生显著更高水平的VEGF、血管生成素-2和骨桥蛋白。15-16月龄小鼠视网膜下纤维化病变比2.5月龄青年小鼠大62%。与年轻小鼠相比，老年小鼠的病变中含有显著更高数量的纤维细胞。视网膜下纤维化病变的大小与循环中纤维细胞的数量呈正相关。老年小鼠骨髓移植显著增加年轻小鼠视网膜下纤维化。在视网膜损伤期间存在纤维细胞/巨噬细胞募集的视网膜-BM-血液-视网膜途径。衰老通过更高数量的循环纤维细胞和BM衍生的巨噬细胞的促纤维化潜力促进视网膜下纤维化。在线版本包含补充材料，可通过10.1186/s12974-023-02731-y获得。
Retinal fibrosis affects 40–70% of neovascular age-related macular degeneration patients. This study investigated the effect of ageing on subretinal fibrosis secondary to choroidal neovascularization and the mechanism of action. Subretinal fibrosis was induced in young (2.5-month) and aged (15–16-month) C57BL/6J mice using the two-stage laser protocol. Five and 30 days later, eyes were collected and stained for CD45 and collagen-1 and observed by confocal microscopy. Fibrocytes (CD45+collagen-1+) were detected in the bone marrow (BM), blood and fibrotic lesions by flow cytometry and confocal microscopy, respectively. BM-derived macrophages (BMDMs) were cultured from young and aged mice with or without TGF-β1 (10 ng/mL) treatment. The expression of mesenchymal marker αSMA (Acta2), fibronectin (Fn1) and collagen-1 (Col1a1) was examined by qPCR and immunocytochemistry, whereas cytokine/chemokine production was measured using the Luminex multiplex cytokine assay. BM were transplanted from 22-month (Ly5.2) aged mice into 2.5-month (Ly5.1) young mice and vice versa. Six weeks later, subretinal fibrosis was induced in recipient mice and eyes were collected for evaluation of fibrotic lesion size. Under normal conditions, the number of circulating fibrocytes (CD45+collagen-1+) and the expression levels of Tgfb1, Col1a1, Acta2 and Fn1 in BMDMs were significantly higher in aged mice compared to young mice. Induction of subretinal fibrosis significantly increased the number of circulating fibrocytes, enhanced the expression of Col1a1, Acta2 and Fn1 and the production of soluble urokinase plasminogen activator surface receptor (suPAR) but decreased the production of CXCL10 in BMDMs. BMDMs from aged subretinal fibrosis mice produced significantly higher levels of VEGF, angiopoietin-2 and osteopontin than cells from young subretinal fibrosis mice. The subretinal fibrotic lesion in 15–16-month aged mice was 62% larger than that in 2.5-month young mice. The lesion in aged mice contained a significantly higher number of fibrocytes compared to that in young mice. The number of circulating fibrocytes positively correlated with the size of subretinal fibrotic lesion. Transplantation of BM from aged mice significantly increased subretinal fibrosis in young mice. A retina–BM–blood–retina pathway of fibrocyte/macrophage recruitment exists during retinal injury. Ageing promotes subretinal fibrosis through higher numbers of circulating fibrocytes and profibrotic potential of BM-derived macrophages. The online version contains supplementary material available at 10.1186/s12974-023-02731-y.
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影响因子：
7.8
作者：
Chen M;Rajapakse D;Fraczek M;Luo C;Forrester JV;Xu H
通讯作者：
Xu H
影响因子：
13.7
作者：
Daniel E;Pan W;Ying GS;Kim BJ;Grunwald JE;Ferris FL 3rd;Jaffe GJ;Toth CA;Martin DF;Fine SL;Maguire MG;Comparison of Age-related Macular Degeneration Treatments Trials
通讯作者：
Comparison of Age-related Macular Degeneration Treatments Trials
影响因子：
3.7
作者：
Chen H;Liu B;Lukas TJ;Neufeld AH
通讯作者：
Neufeld AH
影响因子：
15.9
作者：
Jiang, Dianhua;Liang, Jiurong;Noble, Paul W.
通讯作者：
Noble, Paul W.
影响因子：
7.4
作者：
Duran-Struuck R;Hartigan A;Clouthier SG;Dyson MC;Lowler K;Gatza E;Tawara I;Toubai T;Weisiger E;Hugunin K;Reddy P;Wilkinson JE
通讯作者：
Wilkinson JE