Peng Jing, Ma Liang, Wang Zhonghui, Du Yaxi, Tao Qunfen, Wang Qiongchuan, Zhao Li
Department of Anesthesiology, Yunnan Cancer Hospital, The Third Affiliated Hospital of Kunming Medical University, Peking University Cancer Hospital Yunnan, Kunming, Yunnan, China.
Department of Molecular Diagnosis Center, Yunnan Cancer Hospital, The Third Affiliated Hospital of Kunming Medical University, Peking University Cancer Hospital Yunnan, Kunming, Yunnan, China.
PLoS One. 2025 Jul 2;20(7):e0325936. doi: 10.1371/journal.pone.0325936. eCollection 2025.
Angiogenic signaling pathway activation has been shown to accelerate compensatory lung growth (CLG) after unilateral pneumonectomy (PNX). Therefore, studying specific genes regulating angiogenic signaling pathways is a novel strategy to promote CLG. EdU, flow cytometry and tube formation experiments were performed to test the metabolism of human pulmonary microvascular endothelial cells (HPMECs). Western blotting was used to analyze the levels of promyelocytic leukemia zinc finger protein (PLZF), kelch-like ECH-associated protein 1 (Keap1), hypoxia-inducible factor-1α (HIF-1α), hemeoxygenase-1 (HO-1), quinone oxidoreductase (NQO1), nuclear factor E2-related factor 2 (Nrf2) and other proteins. The proliferation of pulmonary endothelial cells was assessed by Ki67 double staining. A unilateral PNX mouse model was constructed, and changes in lung volume and weight were assessed. Our bioinformatics results suggested that PLZF showed a clear downward trend after unilateral PNX. PLZF overexpression significantly promoted HPMECs proliferation and angiogenesis and inhibited their apoptosis. Further studies revealed that both Keap1 overexpression and Nrf2 silencing altered the effects of PLZF overexpression on HPMECs and inhibited their apoptosis. Notably, HIF-1α silencing reversed the effect of PLZF overexpression on HPMECs angiogenesis but not on proliferation or apoptosis. Knockdown of Nrf2 not only affected HPMECs proliferation and apoptosis but also affected angiogenesis. An in vivo study confirmed that PLZF overexpression promoted an increase in residual lung volume and lung weight in mice after unilateral PNX and significantly promoted the proliferation of lung endothelial cells. In conclusion, our study revealed that PLZF promotes HPMECs proliferation and angiogenesis and accelerates CLG by inhibiting Keap1 activation of the Nrf2 and HIF-1α/VEGF signaling pathways.
血管生成信号通路的激活已被证明可加速单侧肺切除术后的代偿性肺生长（CLG）。因此，研究调节血管生成信号通路的特定基因是促进CLG的一种新策略。进行EdU、流式细胞术和管形成实验以检测人肺微血管内皮细胞（HPMECs）的代谢。采用蛋白质免疫印迹法分析早幼粒细胞白血病锌指蛋白（PLZF）、kelch样ECH相关蛋白1（Keap1）、缺氧诱导因子-1α（HIF-1α）、血红素加氧酶-1（HO-1）、醌氧化还原酶（NQO1）、核因子E2相关因子2（Nrf2）等蛋白的水平。通过Ki67双重染色评估肺内皮细胞的增殖。构建单侧肺切除小鼠模型，并评估肺体积和重量的变化。我们的生物信息学结果表明，单侧肺切除后PLZF呈明显下降趋势。PLZF过表达显著促进HPMECs增殖和血管生成，并抑制其凋亡。进一步研究表明，Keap1过表达和Nrf2沉默均改变了PLZF过表达对HPMECs的影响，并抑制其凋亡。值得注意的是，HIF-1α沉默逆转了PLZF过表达对HPMECs血管生成的影响，但对增殖或凋亡无影响。Nrf2敲低不仅影响HPMECs增殖和凋亡，还影响血管生成。体内研究证实，PLZF过表达促进单侧肺切除术后小鼠残余肺体积和肺重量增加，并显著促进肺内皮细胞增殖。总之，我们的研究表明，PLZF通过抑制Keap1对Nrf2和HIF-1α/VEGF信号通路的激活，促进HPMECs增殖和血管生成，并加速CLG。
