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Experimental cell research

Experimental cell research

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The RPE-derived NF-κB/HO-1/MCP-1 pathway mediated Microglia Recruitment Involved in the Outer Blood-Retinal Barrier Breakdown in Experimental Diabetic Retinopathy

Published:30 October 2025 DOI: 10.1016/j.yexcr.2025.114813 PMID: 41176205
Yiyang Shu , Dandan Liu , Hai Xie , Chaoyang Zhang , Yanlong Bi , Jingfa Zhang

Abstract

Purpose

To investigate the mechanisms underlying the microglia recruitment and its causal role in the breakdown of the outer blood-retinal barrier (oBRB) in diabetic retinopathy (DR).

Methods

The Sprague-Dawley rats were adopted to establish diabetic model by intraperitoneal injection of streptozotocin. Twelve weeks later, the retinal pigment epithelium (RPE)-choroid complexes and retinal paraffin sections were examined with immunofluorescence. RNA-sequencing was performed on glyoxal-treated ARPE-19?cells, followed by bioinformatic analysis to identify significant genes and pathways. Transwell assays were employed to establish the co-culture system and investigate the interactions between ARPE-19 and BV2 microglial cells. The results were further validated by the inhibitor or siRNAs targeting NF-κB, HO-1, and MCP-1.

Results

In 12-week diabetic rat retinas, microglia were observed to accumulate in the vicinity of the RPE cells, accompanied by the disruption of ZO-1. The expressions of ZO-1 and occludin remained largely unchanged in ARPE-19?cells when treated with glyoxal alone. However, when co-cultured with BV2 microglial cells, the expression levels of ZO-1 and occludin in glyoxal-treated ARPE-19?cells were significantly decreased, which were effectively prevented by siMCP-1. Mechanistically, RNA-sequencing analysis revealed that the activation of the NF-κB/HO-1/MCP-1 pathway in glyoxal-treated ARPE-19?cells significantly contributed to the recruitment of microglia. The above effects were reversed by BAY 11–7082, siHO-1 or siMCP-1.

Conclusion

Under diabetic conditions, microglia are recruited by RPE cells via the NF-κB/HO-1/MCP-1 pathway, which subsequently results in the oBRB breakdown. This study provides a novel mechanistic insight for the interaction between microglia and RPE cells, and implies a potential therapeutic strategy for the treatment of DR.

Substances (8)

Materials
Procduct Name CAS Molecular Formula Supplier Price
BAY 11-7082 19542-67-7 C10H9NO2S 253 suppliers $14.00-$2304.00
BAY 11-7082 19542-67-7 C10H9NO2S 253 suppliers $14.00-$2304.00
BAY 11-7082 19542-67-7 C10H9NO2S 253 suppliers $14.00-$2304.00
BAY 11-7082 19542-67-7 C10H9NO2S 253 suppliers $14.00-$2304.00
HO-1-IN-1 1093058-52-6 C13H15BrN2 28 suppliers Inquiry
HO-1-IN-1 1093058-52-6 C13H15BrN2 28 suppliers Inquiry
HO-1-IN-1 1093058-52-6 C13H15BrN2 28 suppliers Inquiry
HO-1-IN-1 1093058-52-6 C13H15BrN2 28 suppliers Inquiry
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