Han Longhui, Su Wenru, Huang Jingwen, Zhou Jingwen, Qiu Sujuan, Liang Dan
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China; Hebei Provincial Key Laboratory of Ophthalmology, Hebei Provincial Eye institute, Hebei Eye Hospital, Xingtai, China.
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
PLoS One. 2014 Sep 30;9(9):e108931. doi: 10.1371/journal.pone.0108931. eCollection 2014.
Lymphangiogenesis is significantly involved in the pathogenesis of diseases, including graft rejection, cancer metastasis and various inflammatory conditions. The inhibition of lymphangiogenesis has become a new therapeutic target for the treatment of these diseases. Here, we explored the anti-lymphangiogenic effects of doxycycline in inflammation-induced lymphangiogenesis (ILA) in the cornea and the underlying mechanisms. In the present study, mice with ILA of the cornea were treated with topical doxycycline (0.1%) or vehicle control. Lymphangiogenesis was quantified using corneal immunostaining of lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1). Human dermal lymphatic endothelial cells (HDLECs) and a murine macrophage cell line (RAW264.7) were used to further explore the underlying mechanisms of doxycycline-mediated anti-lymphangiogenesis in vitro. Our results showed that doxycycline treatment dramatically inhibited ILA in the mouse cornea (p<0.001), with a significant decrease in vascular endothelial growth factor (VEGF)-C/VEGF receptor 3 signalling, macrophage infiltration and inflammatory cytokine expression. Doxycycline also significantly inhibited VEGF-C-induced HDLEC proliferation in vitro by modulating the PI3K/Akt/endothelial nitric oxide (NO) synthase (eNOS) pathway and significantly suppressed interleukin-1β (IL-1β), TNF-α and VEGF-C production in the RAW264.7 cell line by modulating the PI3K/Akt/nuclear factor-kappaB (NF-κB) pathway. Additionally, doxycycline treatment dramatically reduced the phosphorylation of NF-κBp65, Akt and eNOS in ILA and significantly inhibited matrix metalloproteinases (MMPs) activity in vitro and in ILA. In conclusion, doxycycline inhibited ILA, possibly through suppression of VEGF-C signalling, macrophage function and MMPs activity. This observation suggests that doxycycline is a potential therapeutic agent for lymphangiogenesis-related diseases.
淋巴管生成在包括移植排斥、癌症转移和各种炎症性疾病在内的疾病发病机制中起着重要作用。抑制淋巴管生成已成为治疗这些疾病的新治疗靶点。在此，我们探讨了强力霉素在角膜炎症诱导的淋巴管生成（ILA）中的抗淋巴管生成作用及其潜在机制。在本研究中，对患有角膜ILA的小鼠局部应用强力霉素（0.1%）或赋形剂对照进行治疗。使用淋巴管内皮透明质酸受体-1（LYVE-1）的角膜免疫染色对淋巴管生成进行定量。使用人真皮淋巴管内皮细胞（HDLEC）和小鼠巨噬细胞系（RAW264.7）在体外进一步探索强力霉素介导的抗淋巴管生成的潜在机制。我们的结果表明，强力霉素治疗显著抑制了小鼠角膜中的ILA（p<0.001），血管内皮生长因子（VEGF）-C/VEGF受体3信号传导、巨噬细胞浸润和炎性细胞因子表达显著降低。强力霉素还通过调节PI3K/Akt/内皮型一氧化氮（NO）合酶（eNOS）途径显著抑制体外VEGF-C诱导的HDLEC增殖，并通过调节PI3K/Akt/核因子-κB（NF-κB）途径显著抑制RAW264.7细胞系中白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）和VEGF-C的产生。此外，强力霉素治疗显著降低了ILA中NF-κBp65、Akt和eNOS的磷酸化，并在体外和ILA中显著抑制基质金属蛋白酶（MMP）活性。总之，强力霉素可能通过抑制VEGF-C信号传导、巨噬细胞功能和MMP活性来抑制ILA。这一观察结果表明，强力霉素是一种治疗淋巴管生成相关疾病的潜在治疗药物。
