Journal of Medical Molecular Biology ›› 2023, Vol. 20 ›› Issue (3): 188-195.doi: 10.3870/j.issn.1672-8009.2023.03.001

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Expression of Biliverdin Reductase During Macrophage Differentiation

  

  1. Division of Nephrology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China
  • Online:2023-05-31 Published:2023-05-29

Abstract: Objective To investigate the expression of biliverdin reductase (BVR) in bone marrow derived macrophages and RAW264. 7 cells. Methods L929 cells were cultured for 2 days, the supernatant was then collected. The murine bone marrow derived macrophage cells were isolated and were cultured with the above supernatant for 7 days. Cells were then divided into three groups: control group, LPS + IFN-γ group, IL-4 + IL-13 group. Cells in the LPS + IFN-γ group and the IL-4 + IL-13 group were stimulated by 100 ng / mL LPS and IFN-γ or 10 ng / mL IL-4 and 10 ng / mL IL-13 for 24 hours respectively. Western blotting was used to detect the expression levels of INOS and ARG-1 proteins. RT-PCR was used to detect the RNA expression levels of INOS, ARG-1, FN and FIZZ1. Flow cytometry was used to detect the expression of CD45, F4 / 80, CD11b, CD86 and DECTIN-1 on cells. . The expression level of BVR in the macrophage polarization model established in BMDM cells and RAW264. 7 cells was detected by RT-PCR. Results ① The protein expression level of INOS was increased in the LPS + IFN-γ group, and the protein expression level of ARG-1 was increased in the IL-4 + IL-13 group. ② The expression of CD86 on cells was increased in the LPS + IFN-γ group, and the expression of DECTIN-1 on cells was increased in the IL-4 + IL-13 group. ③ The RNA expression levels of INOS and TNF-α were increased in the LPS + IFN-γ group, the RNA expression levels of ARG-1, FN and FIZZ1 were increased in IL-4 + IL-13 group. ④ The expression level of BVR was increased in the M2-shift macrophages, but decreased in the M1-shift macrophages in BMDM and RAW264. 7 cell models. Conclusion ① The macrophage polarization models are successfully established by stimulation of LPS and IFN-γ or IL-4 and IL-13 in vitro. ② The expression level of BVR is increased in the M2-shift macrophages, but decreased in the M1- shift macrophages in vitro.

Key words: macrophage, biliverdin reductase, differentiation

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