MicroRNA‑216a‑5p in lipopolysaccharide‑induced endothelial injury

Liu,Wenxun; Xi,Wenhua; Li,Yan; Hai,Kerong; Zhou,Xiaohong; Wang,Yun; Ye,Qingshan

doi:10.3892/etm.2021.10861

MicroRNA‑216a‑5p in lipopolysaccharide‑induced endothelial injury

Authors:
- Wenxun Liu
- Wenhua Xi
- Yan Li
- Kerong Hai
- Xiaohong Zhou
- Yun Wang
- Qingshan Ye
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Affiliations: Department of Anesthesiology, Ningxia Medical University, Yinchuan, Gansu 750004, P.R. China, Department of Anesthesiology, People's Hospital of Ningxia Hui Autonomous Region, Yinchuan, Ningxia 750002, P.R. China
Published online on: October 11, 2021 https://doi.org/10.3892/etm.2021.10861
Article Number: 1426
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

MicroRNAs (miRNAs/miRs) are a type of non‑coding RNA that are closely associated with disease development and treatment. The present study aimed to investigate the role of miR‑216a‑5p in lipopolysaccharide (LPS)‑induced endothelial injury in vitro. The EdU assay was performed to detect EdU‑positive cells, while flow cytometric analysis was performed to detect apoptotic cells. Reverse transcription‑quantitative PCR and western blot analyses were performed to detect the expression levels of miR‑216a‑5p, Toll‑like receptor 4 (TLR4), MyD88 and nuclear factor (NF)‑κB(p65) and phosphorylated (p)‑NF‑κB(p65). Furthermore, p‑NF‑κB(p65) nuclear expression level was detected via cellular immunofluorescence. The dual‑luciferase reporter assay was performed to verify the association between miR‑216a‑5p and TLR4. The results demonstrated that the number of EdU‑positive cells significantly decreased, the apoptotic rate significantly increased, and TLR4, MyD88 and NF‑κB(p65) mRNA expression levels were significantly upregulated.TLR4, MyD88 and p‑NF‑κB(p65) protein expression levels were significantly upregulated and p‑NF‑κB(p65) nuclear concentration was significantly enhanced in the small interfering RNA‑miR‑216a‑5p and LPS groups (P<0.001, respectively) compared with the negative control group. However, the addition of miR‑216a‑5p significantly increased the number of EdU‑positive cells, significantly decreased the apoptotic rate and significantly downregulated the mRNA expression levels of TLR4, MyD88 and NF‑κB(p65), as well as the protein expression levels of TLR4, MyD88 and p‑NF‑κB(p65). In addition, the p‑NF‑κB(p65) nuclear concentration was significantly decreased in the miR‑216a‑5p group (P<0.001, respectively) compared with the LPS group. Taken together, the results suggest that overexpression of miR‑216a‑5p suppresses the effects of LPS induced endothelial injury.

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