PMID: 29773101May 19, 2018Paper

Microvesicles derived from LPS-induced microglia aggravate the injury of tight junction in rat brain microvascular endothelial cells under oxygen-glucose deprivation

Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Yanni LyuYisong Qian

Abstract

Objective To study the effect of the administration of the circulating microvesicles (MV) obtained from lipopolysaccharide (LPS)-stimulated microglia supernatant into rat brain endothelial cells (RBECs) on the injury of tight junction in RBECs under the condition of oxygen-glucose deprivation (OGD) as well as the underlying mechanism. Methods The circulating MV was isolated from the supernatant of microglia stimulated with LPS 1 mg/L for 24 hours and subjected to morphological identification. The expression of miR-27a in MV was detected by real-time PCR. RBECs were randomly divided into control group, MV-RBECs control group, OGD 6-hour group and OGD-MV group. The expressions of occludin and claudin-5 were detected by immunofluorescence staining in the four groups. Western blot analysis was used to investigate the expressions of occludin, claudin-5, Toll-like receptor 4 (TLR4), NF-κBp65 and p38 proteins in RBECs, and ELISA was applied to detect the levels of interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) in RBECs. Results The shape of MV was approximately circular double membrane vesicles, with an average diameter of 150 nm, in accordance with the morphological characteristics of MV. Under LPS stimulation, the level ...Continue Reading

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