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Dexamethasone and MicroRNA-204 Inhibit Corneal Neovascularization

Xiaoping Zhang¹, Gang Wang², Qing Wang¹

¹Department of Ophthalmology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266002, China.

Military Medicine

|August 31, 2022

Abstract

Introduction:

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.

Materials And Methods:

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.

Results:

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).

Conclusion:

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.

Insights

Dexamethasone (Dex) may reduce corneal neovascularization (CNV) by increasing microRNA-204 (miR-204). This study found miR-204 inhibits cell migration and tube formation, suggesting it is a therapeutic target for alkali burn-induced CNV.

Area of Science:

Ophthalmology
Molecular Biology
Pharmacology

Background:

Corneal neovascularization (CNV) is a significant cause of vision impairment.
Alkali burns are a common cause of severe ocular surface injury leading to CNV.
Dexamethasone (Dex) is a corticosteroid used to treat ocular inflammation, but its precise mechanism in CNV is not fully understood.

Purpose of the Study:

To investigate the interaction between dexamethasone (Dex) and microRNA-204 (miR-204) in a mouse model of alkali burn-induced corneal neovascularization (CNV).
To elucidate the functional role of miR-204 in human mammary epithelial cells (HMECs) in vitro.
To explore miR-204 as a potential therapeutic target for CNV.

Main Methods:

Established a corneal alkali burn model in BLAB/c mice.
Administered subconjunctival injections of Dex, negative control (NTC), or miR-204 agomir.
Quantified CNV area using immunofluorescence staining and miR-204 expression via qRT-PCR.
Assessed HMEC migration and tube formation in vitro after miR-204 modulation.

Main Results:

Alkali burn significantly reduced miR-204 expression.
Dex treatment upregulated miR-204 expression and reduced CNV area.
miR-204 agomir significantly inhibited HMEC migration and tube formation in vitro.
miR-204 antagomir demonstrated opposite effects, accelerating HMEC tube formation.

Conclusions:

Dexamethasone's anti-CNV effect is partly mediated by the upregulation of miR-204.
MicroRNA-204 plays an inhibitory role in corneal neovascularization.
miR-204 represents a promising therapeutic target for managing alkali burn-induced CNV.

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