胎球蛋白-B 过表达通过激活小胶质细胞和 NF-κB 信号通路促进糖尿病视网膜病变中的炎症反应。

Fetuin-B Overexpression Promotes Inflammation in Diabetic Retinopathy Through Activating Microglia and the NF-κB Signaling Pathway.

机构信息

Department of Ophthalmology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.

School of Biomedical Sciences, Faculty of Medicine, The University of Queensland, Brisbane, QLD, Australia.

出版信息

Curr Eye Res. 2024 Feb;49(2):168-179. doi: 10.1080/02713683.2023.2276683. Epub 2024 Jan 18.

DOI:10.1080/02713683.2023.2276683

PMID:37883127

Abstract

PURPOSE

To investigate the expression, source, role, and mechanism of Fetuin-B (FETUB) in diabetic retinopathy (DR).

METHODS

ELISA and immunofluorescence were used to analyze the concentration of FETUB in plasma, aqueous fluid, and tissue specimens of patients with DR and healthy controls. Immunofluorescence, q-PCR, and western blotting were used to examine the expression of FETUB in DR mice and cells cultured with different concentrations of glucose. BV2 microglia cell line and DR mice were treated using FETUB recombination protein and FETUB shRNA to explore the function of FETUB in DR by q-PCR, western blotting, and immunofluorescence.

RESULTS

FETUB concentrations in plasma, aqueous fluid, and tissue specimens were significantly increased in DR patients. The mice in DR group had a higher concentration of FETUB in the retina and liver tissues than those in the control group, and the expression of FETUB was increased in both ARPE19 and BV2 cells under a high-glucose environment. The ratio of p-P65 (Phospho-P65)/P65 and the expression levels of TNF-α, VEGF, and ionized calcium binding adaptor molecule (IBA)-1 were increased in BV2 cells cultured with FETUB recombinant protein, while they were decreased in BV2 cells transfected with FETUB shRNA. Immunofluorescence staining showed that there were more IBA-1 activated microglia in the retinas of the FETUB recombination protein group than in the retinas of the DR group, and there were fewer IBA-1 activated microglia in the retinas of the FETUB shRNA group than in the retinas of the DR group.

CONCLUSIONS

FETUB sourced from endocrine, autocrine, and paracrine pathways could promote inflammation in DR by activating the NF-κB pathway and microglia.

摘要

目的

研究胎球蛋白-B（FETUB）在糖尿病视网膜病变（DR）中的表达、来源、作用和机制。

方法

采用 ELISA 和免疫荧光法分析 DR 患者和健康对照者血浆、房水和组织标本中 FETUB 的浓度。采用免疫荧光、q-PCR 和 Western blot 法检测 DR 小鼠和不同浓度葡萄糖培养的细胞中 FETUB 的表达。采用 FETUB 重组蛋白和 FETUB shRNA 处理 BV2 小胶质细胞系和 DR 小鼠，通过 q-PCR、Western blot 和免疫荧光法探讨 FETUB 在 DR 中的作用。

结果

DR 患者血浆、房水和组织标本中 FETUB 浓度明显升高。DR 组小鼠视网膜和肝脏组织中 FETUB 浓度高于对照组，高糖环境下 ARPE19 和 BV2 细胞中 FETUB 表达增加。FETUB 重组蛋白处理的 BV2 细胞中 p-P65（磷酸化-P65）/P65 比值和 TNF-α、VEGF、离子钙结合衔接分子（IBA）-1 的表达水平升高，而 FETUB shRNA 转染的 BV2 细胞中则降低。免疫荧光染色显示，FETUB 重组蛋白组小鼠视网膜中 IBA-1 活化的小胶质细胞较 DR 组增多，而 FETUB shRNA 组小鼠视网膜中 IBA-1 活化的小胶质细胞较 DR 组减少。