Abstract
The present study aimed to determine the expression level of claudin-2 (CLDN2) in the peripheral blood of patients with sepsis, and to investigate its potential function and mechanism of action in vascular endothelial injury. A total of 25 patients with sepsis were included in the present study. Reverse transcription-quantitative PCR was used to determine CLDN2 levels in peripheral blood. HUVECs stably expressing CLDN2 were prepared and Cell Counting Kit-8, flow cytometry and Transwell assays were performed to study the proliferation, apoptosis and migration of HUVECs, respectively. Using bioinformatics, microRNA (miR) molecules that interact with CLDN2 were predicted. A dual luciferase reporter assay was used to test whether miR-331 regulated CLDN2. Western blotting was employed to determine CLDN2 protein expression. In addition, in vitro transfection of HUVECs with miR-331 mimics was performed to test the rescue effects of miR-331 on the cell function changes induced by CLDN2. The results indicated that elevated CLDN2 expression altered the proliferation and cell cycle of peripheral vascular endothelial cells. CLDN2 overexpression inhibited HUVEC proliferation via mechanisms not associated with the cell cycle. CLDN2 mRNA levels in the peripheral blood of patients with sepsis were significantly higher than those in healthy subjects. Upregulated CLDN2 expression promoted the apoptosis of HUVECs, but reduced their proliferation and migration. Notably, miR-331 was able to bind with CLDN2 mRNA and regulate its expression. Upregulation of miR-331 expression inhibited the expression of CLDN2 and restored nearly normal proliferation, apoptosis and migration to HUVECs. The present study demonstrated that CLDN2 expression is elevated in peripheral blood from patients with sepsis, and promotes the injury of vascular endothelial cells. In addition, miR-331 participates in the direct regulation of CLDN2, and upregulation of miR-331 expression inhibits the expression of CLDN2 and restores cellular functions to HUVECs.