Heparin alleviates LPS‑induced endothelial injury by regulating the TLR4/MyD88 signaling pathway

Liu,Wenxun; Li,Yan; Wu,Zhaozhao; Hai,Kerong; Wang,Yun; Zhou,Xiaohong; Ye,Qingshan

doi:10.3892/etm.2021.10833

Heparin alleviates LPS‑induced endothelial injury by regulating the TLR4/MyD88 signaling pathway

Authors:
- Wenxun Liu
- Yan Li
- Zhaozhao Wu
- Kerong Hai
- Yun Wang
- Xiaohong Zhou
- Qingshan Ye
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Affiliations: Anesthesia Specialty, Ningxia Medical University, Yinchuan, Ningxia 750004, P.R. China, Department of Anesthesiology, Northwest Minzu University, Lanzhou, Gansu 730030, P.R. China, Department of Anesthesiology, People's Hospital of Ningxia Hui Autonomous Region, Yinchuan, Ningxia 750002, P.R. China
Published online on: October 1, 2021 https://doi.org/10.3892/etm.2021.10833
Article Number: 1397
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Heparin is a commonly used in the clinic, however, Heparin's effect on endothelial injury remains unclear. The aim of the present study was to evaluate the effects and possible mechanisms of action underlying heparin treatment in lipopolysaccharide (LPS)‑induced endothelial injury in vitro. TNF‑α, IL‑1β, IL‑6 and IFN‑γ levels were measured using ELISA. Cell proliferation was measured using a 5‑ethynyl‑2'‑deoxyuridine (EdU) assay. The number of apoptotic cells and apoptotic rate were evaluated using TUNEL assays and flow cytometry, respectively. Toll‑like receptor 4 (TLR4), myeloid differentiation primary response 88 (MyD88) and NF‑κB (p65) gene expression was evaluated using reverse transcription‑quantitative PCR, whilst TLR4, MyD88 and p‑NF‑κB (p65) protein expression was evaluated using western blot analysis. The levels of phosphorylated NF‑κB in the nucleus were evaluated using cellular immunofluorescence. Compared with those in the normal control group, TNF‑α, IL‑1β, IL‑6 and IFN‑γ levels were significantly increased in the LPS group (P<0.001). In addition, 5‑ethynyl‑2'‑deoxyuridine (EdU)‑positive cells were significantly increased and apoptosis was significantly decreased (P<0.001). TLR4, MyD88 and NF‑κB (p65) expression was also significantly increased (P<0.001). Compared with those in the LPS group, following heparin treatment, TNF‑α, IL‑1β, IL‑6 and IFN‑γ levels were significantly decreased (P<0.05), whilst the number of EdU‑positive cells was significantly increased and the level of apoptosis was significantly decreased (P<0.05). TLR4, MyD88 and NF‑κB (p65) expression was also significantly decreased by heparin in a dose‑dependent manner (P<0.001). Small interfering RNA‑TLR4 transfection exerted similar effects to those mediated by heparin in alleviating endothelial injury. In conclusion, heparin suppressed LPS‑induced endothelial injury through the regulation of TLR4/MyD88/NF‑κB (p65) signaling in vitro.

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