Macrophage exosomes transfer angiotensin II type 1 receptor to lung fibroblasts mediating bleomycin-induced pulmonary fibrosis.
Macrophage exosomes transfer angiotensin II type 1 receptor to lung fibroblasts mediating bleomycin-induced pulmonary fibrosis.
复制标题
巨噬细胞外泌体将血管紧张素 II 1 型受体转移至肺成纤维细胞介导博莱霉素诱导的肺纤维化
DOI：
10.1097/cm9.0000000000001605
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发表时间：
2021-09-02
影响因子：
6.1
通讯作者：
Meng Y
中科院分区：
文献类型：
--
作者：
Sun NN;Zhang Y;Huang WH;Zheng BJ;Jin SY;Li X;Meng Y
巨噬细胞参与特发性肺纤维化的发病机制，部分通过激活肺成纤维细胞。然而，巨噬细胞如何与肺成纤维细胞沟通在很大程度上尚未探索。外泌体可以介导细胞间通讯，但其在肺纤维化中的作用尚不清楚。在这里，我们的目的是研究外泌体是否可以介导巨噬细胞和肺成纤维细胞之间的串扰，并随后诱导纤维化。在体内，建立博莱霉素（BLM）诱导的肺纤维化模型，并检查巨噬细胞浸润。评估了外来体抑制剂GW 4869对肺纤维化的影响。此外，将巨噬细胞外泌体注射到小鼠中以观察其促纤维化作用。在体外，收集来源于血管紧张素II（Ang II）刺激的巨噬细胞的外来体。然后，用外来体处理肺成纤维细胞。24小时后，检测肺成纤维细胞中α-胶原I、血管紧张素II 1型受体（AT 1 R）、转化生长因子-β（TGF-β）和磷酸化Smad 2/3（p-Smad 2/3）的蛋白水平。使用Student t检验或方差分析进行统计分析。在体内，BLM治疗的小鼠表现出增强的巨噬细胞浸润，增加纤维化的变化，和更高水平的血管紧张素II和AT 1 R。GW 4869减弱BLM诱导的肺纤维化。注射外泌体的小鼠表现出纤维化特征，血管紧张素II和血管紧张素1受体水平升高，厄贝沙坦可逆转。在体外实验中，我们发现巨噬细胞分泌大量的外泌体。外泌体被成纤维细胞摄取，导致AT 1 R水平升高。（0.22 ± 0.02 vs.0.07 ± 0.02，t = 8.66，P = 0.001），TGF-β（0.54 ± 0.05 vs.0.09 ± 0.06，t = 10.00，P <0.001），p-Smad 2/3（0.58 ± 0.06 vs.0.07 ± 0.03，t = 12.86，P <0.001）和α-胶原I（0.27 ± 0.02 vs. 0.16 ± 0.01，t = 7.01，P = 0.002），Ang Ⅱ分泌增加（62.27 ± 7.32vs9.56 ± 1.68，t = 12.16，P <0.001）。 有趣的是，Ang II增加了巨噬细胞外泌体的数量，并且在相同数量的巨噬细胞分泌的外泌体中，阿利克斯（1.45± 0.15 vs. 1.00 ± 0.10，t = 4.32，P = 0.012）、AT 1 R（4.05 ± 0.64 vs. 1.00 ± 0.09，t = 8.17，P = 0.001）和甘油醛-3-磷酸脱氢酶（2.13 ± 0.36 vs. 1.00 ± 0.10，t = 5.28，P =0.006）的蛋白水平增加，表明Ang II和外泌体产生之间的正循环。 外泌体介导巨噬细胞和成纤维细胞之间的细胞间通讯在BLM诱导的肺纤维化中起重要作用。
Macrophages are involved in the pathogenesis of idiopathic pulmonary fibrosis, partially by activating lung fibroblasts. However, how macrophages communicate with lung fibroblasts is largely unexplored. Exosomes can mediate intercellular communication, whereas its role in lung fibrogenesis is unclear. Here we aim to investigate whether exosomes can mediate the crosstalk between macrophages and lung fibroblasts and subsequently induce fibrosis. In vivo, bleomycin (BLM)-induced lung fibrosis model was established and macrophages infiltration was examined. The effects of GW4869, an exosomes inhibitor, on lung fibrosis were assessed. Moreover, macrophage exosomes were injected into mice to observe its pro-fibrotic effects. In vitro, exosomes derived from angiotensin II (Ang II)-stimulated macrophages were collected. Then, lung fibroblasts were treated with the exosomes. Twenty-four hours later, protein levels of α-collagen I, angiotensin II type 1 receptor (AT1R), transforming growth factor-β (TGF-β), and phospho-Smad2/3 (p-Smad2/3) in lung fibroblasts were examined. The Student's t test or analysis of variance were used for statistical analysis. In vivo, BLM-treated mice showed enhanced infiltration of macrophages, increased fibrotic alterations, and higher levels of Ang II and AT1R. GW4869 attenuated BLM-induced pulmonary fibrosis. Mice with exosomes injection showed fibrotic features with higher levels of Ang II and AT1R, which was reversed by irbesartan. In vitro, we found that macrophages secreted a great number of exosomes. The exosomes were taken by fibroblasts and resulted in higher levels of AT1R (0.22 ± 0.02 vs. 0.07 ± 0.02, t = 8.66, P = 0.001), TGF-β (0.54 ± 0.05 vs. 0.09 ± 0.06, t = 10.00, P < 0.001), p-Smad2/3 (0.58 ± 0.06 vs. 0.07 ± 0.03, t = 12.86, P < 0.001) and α-collagen I (0.27 ± 0.02 vs. 0.16 ± 0.01, t = 7.01, P = 0.002), and increased Ang II secretion (62.27 ± 7.32 vs. 9.56 ± 1.68, t = 12.16, P < 0.001). Interestingly, Ang II increased the number of macrophage exosomes, and the protein levels of Alix (1.45 ± 0.15 vs. 1.00 ± 0.10, t = 4.32, P = 0.012), AT1R (4.05 ± 0.64 vs. 1.00 ± 0.09, t = 8.17, P = 0.001), and glyceraldehyde-3-phosphate dehydrogenase (2.13 ± 0.36 vs. 1.00 ± 0.10, t = 5.28, P = 0.006) were increased in exosomes secreted by the same number of macrophages, indicating a positive loop between Ang II and exosomes production. Exosomes mediate intercellular communication between macrophages and fibroblasts plays an important role in BLM-induced pulmonary fibrosis.