Vitamin D reduces autophagy by regulating NF-κB resistance to Aspergillus fumigatus infection

Gene. 2020 Aug 30;753:144819. doi: 10.1016/j.gene.2020.144819. Epub 2020 May 30.

Abstract

Vitamin D is one of the indispensable nutrients of human body. When vitamin D is deficient, it can cause a series of related diseases, such as respiratory tract infection. The regulatory role of vitamin D in inflammatory immune response and defense has attracted more and more attention. However, few studies have shown that vitamin D regulates inflammation and autophagy in Aspergillus fumigatus infected lungs. In this study, we will explain the mechanism of vitamin D regulating inflammation and autophagy in Aspergillus fumigatus infected lungs.

Methods: Different concentrations of Aspergillus fumigatus spores were injected into mice with deficien diets (VitD-) or sufficient vitamin D (VitD + ) , and the survival rates were recorded. Then, the weight changes of rats were measured every other time. At the same time, a gauze was used to filter the lapped lung tissue to get the pulmonary spores and measured the amount of the spores. The mice with the same concentration of Aspergillus fumigatus infected were cut off and the lung tissue for pathological examination in the deficien diets (VitD-) group or sufficient vitamin D (VitD + ) group. Moreover, the expression of inflammation related factors TNF-α, IL-1β, IL-6 in lung was measured by immunohistochemical method. The expression of TNF-α, IL-1β, IL-6 in the serum of vitamin D deficiency and sufficient mice were measured by ELISA. In vitro, we obtained macrophages from healthy mice and mixed cultures of Aspergillus fumigatus spores and lung macrophages in medium with or without vitamin D. After the cells were infected with Aspergillus fumigatus spores, the expressions of NF-κB and IL-8 were detected by RT-PCR and Western blot. The RAW264.7 cells transfected with GFP-LC3BII were mixed with Aspergillus fumigatus spores, and the expression of cell fluorescence was observed by the fluorescence microscope with or without chloroquine and rapamycin , and the autophagy flow of the cells was measured by Western blot. In the RAW264.7 cells, Lentivirus transfection and SiRNA technologies were used to enhance or reduce the expression of the NF-κB gene (siNF-κB) for investgating the influence of high or low expression of NF-κB in the autophagic flow of vitamin D + or vitamin D-treated RAW264.7 cells.

Results: The survival rate of vitamin D deficient mice infected Aspergillus fumigatus was significantly lower than that of vitamin D sufficient mice, while the number of spores, spore activity, pathological changes of lungs and inflammation in the lungs of vitamin D deficient mice were more severe than that of vitamin D sufficient mice. In vitro cell experiments, when cell was stimulated with vitamin D, the expressions of NF-κB and IL-8 in cells were lower. The autophagic flux and TNF-α, IL-1β, IL-6 and LC3BII in vitamin D group were significantly lower than those in vitamin D deficiency group.

Conclusions: Vitamin D deficiency can aggravate the inflammatory damage in the lungs of Aspergillus fumigatus. When the body is sufficient in vitamin D, if the lungs infect Aspergillus fumigatus spores, the body may resist the infection of Aspergillus fumigatus by reducing the expression of NF-κB, inflammatory factors and autophagy.

Keywords: Aspergillus fumigatus; Autophagy; Inflammatory factors; NF-κB; Vitamin D.

MeSH terms

  • Animals
  • Aspergillosis / immunology
  • Aspergillosis / metabolism
  • Aspergillus fumigatus / immunology
  • Aspergillus fumigatus / pathogenicity
  • Aspergillus fumigatus / physiology
  • Autophagy / drug effects*
  • Cytokines / genetics
  • Interleukin-1beta / metabolism
  • Interleukin-6 / metabolism
  • Lung / metabolism
  • Macrophages / metabolism
  • Macrophages, Alveolar / metabolism
  • Male
  • Mice
  • Mice, Inbred BALB C
  • NF-kappa B / drug effects*
  • NF-kappa B / metabolism
  • RAW 264.7 Cells
  • Tumor Necrosis Factor-alpha / metabolism
  • Vitamin D / metabolism*
  • Vitamin D Deficiency / metabolism

Substances

  • Cytokines
  • Interleukin-1beta
  • Interleukin-6
  • NF-kappa B
  • Tumor Necrosis Factor-alpha
  • Vitamin D