Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266002, China.
Department of Radiology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266002, China.
Mil Med. 2024 Jan 23;189(1-2):374-378. doi: 10.1093/milmed/usac260.
This was an in vivo animal study designed to investigate the interaction between dexamethasone (Dex) and microRNA-204 (miR-204) in a mouse alkali burn-induced corneal neovascularization (CNV) model. The function of miR-204 was then investigated in human mammary epithelial cells (HMECs) in vitro.
The CNV model was induced by corneal alkali burn in BLAB/c mice. The mice were randomly divided into five groups: normal control (Ctrl), alkali burn-induced corneal injury (Alkali), alkali burn + Dex (Dex), alkali burn + negative control (NTC), and alkali burn + miR-204 agomir (miR-204). Subconjunctival injection of NTC, Dex, or miR-204 agomir was conducted at 0, 3, and 6 days, respectively, after alkali burn. The corneas were collected at day 7 after injury, and the CNV area was observed using immunofluorescence staining. The expression of miR-204 was analyzed with quantitative real time (qRT)-PCR. In HMECs, exogenous miR-204 agomir or antagomir was used to strengthen or inhibit the expression of miR-204. Migration assays and tube formation studies were conducted to evaluate the function of miR-204 on HMECs.
At 7 days post-alkali burn, CNV grew aggressively into the cornea. MicroRNA-204 expression was reduced in the Alkali group in contrast with the Ctrl group (P = .003). However, miR-204 was upregulated in the Dex group (vs. alkali group, P = .008). The CNV areas in the NTC and miR-204 groups were 59.30 ± 8.32% and 25.60 ± 2.30%, respectively (P = .002). In vitro, miR-204 agomir showed obvious inhibition on HMEC migration in contrast with NTC (P = .033) and miR-204 antagomir (P = .017). Compared with NTC, miR-204 agomir attenuated tube formation, while miR-204 antagomir accelerated HMEC tube formation (P < .05).
The role of Dex in attenuating CNV may be partly attributed to miR-204. MiR-204 may be a potential therapeutic target in alkali burn-induced CNV.
本研究旨在探讨地塞米松（Dex）与微小 RNA-204（miR-204）在小鼠碱烧伤诱导的角膜新生血管化（CNV）模型中的相互作用，该研究为体内动物实验。此外，我们还在体外对人类乳腺上皮细胞（HMECs）中的 miR-204 功能进行了研究。
通过角膜碱烧伤诱导 BLAB/c 小鼠的 CNV 模型。将小鼠随机分为五组：正常对照组（Ctrl）、碱烧伤诱导的角膜损伤组（Alkali）、碱烧伤+地塞米松组（Dex）、碱烧伤+阴性对照组（NTC）和碱烧伤+miR-204 激动剂组（miR-204）。在碱烧伤后 0、3 和 6 天，分别对 NTC、Dex 或 miR-204 激动剂进行结膜下注射。在损伤后第 7 天收集角膜，并用免疫荧光染色观察 CNV 面积。通过实时定量 PCR（qRT-PCR）分析 miR-204 的表达。在 HMECs 中，用外源性 miR-204 激动剂或拮抗剂增强或抑制 miR-204 的表达。进行迁移实验和管形成实验，以评估 miR-204 对 HMECs 的功能。
在碱烧伤后 7 天，CNV 迅速侵入角膜。与对照组相比，miR-204 在 Alkali 组的表达降低（P=0.003）。然而，在 Dex 组中，miR-204 上调（与碱组相比，P=0.008）。NTC 和 miR-204 组的 CNV 面积分别为 59.30±8.32%和 25.60±2.30%（P=0.002）。在体外，miR-204 激动剂与 NTC（P=0.033）和 miR-204 拮抗剂（P=0.017）相比，对 HMEC 迁移有明显的抑制作用。与 NTC 相比，miR-204 激动剂减弱了管形成，而 miR-204 拮抗剂加速了 HMEC 管形成（P<0.05）。
Dex 减轻 CNV 的作用可能部分归因于 miR-204。miR-204 可能是碱烧伤诱导的 CNV 的潜在治疗靶点。
