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Anti-inflammatory effects of Chinese propolis in lipopolysaccharide-stimulated human umbilical vein endothelial cells by suppressing autophagy and MAPK/NF-κB signaling pathway

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Abstract

This study aimed to investigate the possible benefits of Chinese poplar propolis (CP) in inhibiting inflammation using vascular endothelial cells (VECs) cultured in a nutrient-rich condition exposed to lipopolysaccharide (LPS). Cell proliferation was detected by sulforhodamine B assay and EdU kit. The production of reactive oxygen species (ROS) and level of mitochondrial membrane potential were determined with fluorescent probe DCHF and JC-1, respectively. Protein expression was examined by immunofluorescence staining and western blotting. The results showed that CP (6.25, 12.5, and 25 μg/mL) significantly reduced LPS-induced cytotoxicity, and when challenged with CP substantially suppressed ROS overproduction and protected mitochondrial membrane potential. CP treatment significantly inhibited autophagy by inhibiting LC3B distribution and accumulation, and elevating the p62 level in an mTOR-independent manner but mainly by suppressing the translocation of p53 from the cytoplasm to the nucleus. Furthermore, CP treatment markedly reduced protein levels of TLR4 at 12 and 24 h and significantly suppressed nuclear translocation of NF-κB p65 from cytoplasm to nucleus. In addition, CP treatment significantly reduced the phosphorylation of JNK, ERK1/2, and p38 MAPK. Our findings demonstrated that CP protects VECs from LPS-induced oxidative stress and inflammation, which might be associated with depressing autophagy and MAPK/NF-κB signaling pathway. The results provided novel insights for the potential use of nutrient-rich propolis against inflammation.

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Abbreviations

CP:

Chinese poplar propolis

DCF:

Dichlorofluorescin

DMEM:

Dulbecco’s Modified Eagle’s Medium

EECP:

Ethanol-extracted Chinese propolis

FBS:

Fetal bovine serum

LPS:

Lipopolysaccharide

SRB:

Sulforhodamine B

ROS:

Reactive oxygen species

PI:

Propidium iodide

VECs:

Vascular endothelial cells

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Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (No. 31201860, 31672499), Shandong Province Higher Educational Science and Technology Program (J16LE21) and the earmarked fund for Modern Agro-industry Technology Research System from the Ministry of Agriculture of China (CARS-44).

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  1. Hongzhuan Xuan and Wenwen Yuan contributed equally to this work.

Authors and Affiliations

  1. School of Life Science, Liaocheng University, Liaocheng, 252059, China

    Hongzhuan Xuan, Wenwen Yuan, Huasong Chang & Minmin Liu

  2. College of Animal Sciences, Zhejiang University, Hangzhou, 310029, China

    Fuliang Hu

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  1. Hongzhuan Xuan
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  2. Wenwen Yuan
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  3. Huasong Chang
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  4. Minmin Liu
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Contributions

HX and FH designed the work, drafted the manuscript, and revised it critically. WY, HC, and ML conducted the work, collected, and analyzed the data. All authors agreed to be accountable for all aspects of the work.

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Correspondence to Hongzhuan Xuan or Fuliang Hu.

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All data supporting our findings are adequately contained within the manuscript.

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The authors declare that they have no competing interests.

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Xuan, H., Yuan, W., Chang, H. et al. Anti-inflammatory effects of Chinese propolis in lipopolysaccharide-stimulated human umbilical vein endothelial cells by suppressing autophagy and MAPK/NF-κB signaling pathway. Inflammopharmacol 27, 561–571 (2019). https://doi.org/10.1007/s10787-018-0533-6

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  • DOI: https://doi.org/10.1007/s10787-018-0533-6

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