Transient downregulation of microRNA-206 protects alkali burn injury in mouse cornea by regulating connexin 43.

Abstract

PURPOSE:

Chemical burn in cornea may cause permanent visual problem or complete blindness. In the present study, we investigated the role of microRNA 206 (miR-206) in relieving chemical burn in mouse cornea.

METHOD:

An alkali burn model was established in C57BL/6 mice to induce chemical corneal injury. Within 72 hours, the transient inflammatory responses in alkali-treated corneas were measured by opacity and corneal neovascularization (CNV) levels, and the gene expression profile of miR-206 was measured by quantitative real-time PCR (qPCR). Inhibitory oligonucleotides of miR-206, miR-206-I, were intrastromally injected into alkali-burned corneas. The possible protective effects of down-regulating miR-206 were assessed by both in vivo measurements of inflammatory responses and in vitro histochemical examinations of corneal epithelium sections. The possible binding of miR-206 on its molecular target, connexin43 (Cx43), was assessed by luciferase reporter (LR) and western blot (WB) assays. Cx43 was silenced by siRNA to examine its effect on regulating miR-206 modulation in alkali-burned cornea.

RESULTS:

Opacity and CNV levels, along with gene expression of miR-206, were all transiently elevated within 72 hours of alkali-burned mouse cornea. Intrastromal injection of miR-206-I into alkali-burned cornea down-regulated miR-206 and ameliorated inflammatory responses both in vivo and in vitro. LR and WB assays confirmed that Cx43 was directly targeted by miR-206 in mouse cornea. Genetic silencing of Cx43 reversed the protective effect of miR-206 down-regulation in alkali-burned cornea.

CONCLUSION:

miR-206, associated with Cx43, is a novel molecular modulator in alkali burn in mouse cornea.

• Authors: Xiaoyan Li, Huanfen Zhou, Weiqiang Tang, Qing Guo, Yan Zhang
• Journal: International journal of clinical and experimental pathology (Int J Clin Exp Pathol) Vol. 8 Issue 3 Pg. 2719-27 ( 2015) ISSN: 1936-2625 [Electronic] United States
• PMID: 26045777 (Publication Type: Journal Article)

Chemical References

• Connexin 43
• GJA1 protein, mouse
• MicroRNAs
• Mirn206 microRNA, mouse
• Oligonucleotides
• RNA, Small Interfering
• Sodium Hydroxide

Topics

• Animals
• Burns, Chemical (genetics, metabolism, pathology, therapy)
• Connexin 43 (genetics, metabolism)
• Cornea (blood supply, metabolism, pathology)
• Corneal Injuries (chemically induced, genetics, metabolism, pathology, therapy)
• Corneal Neovascularization (chemically induced, genetics, metabolism, pathology, therapy)
• Corneal Opacity (chemically induced, genetics, metabolism, pathology, prevention & control)
• Disease Models, Animal
• Down-Regulation
• HEK293 Cells
• Humans
• Injections, Intraocular
• Keratitis (chemically induced, genetics, metabolism, pathology, prevention & control)
• Male
• Mice, Inbred C57BL
• MicroRNAs (genetics, metabolism)
• Oligonucleotides (administration & dosage, genetics, metabolism)
• RNA Interference
• RNA, Small Interfering (genetics, metabolism)
• Sodium Hydroxide
• Time Factors
• Transfection
• Wound Healing