2. Academic Validation
  3. Progranulin inhibits LPS-induced macrophage M1 polarization via NF-кB and MAPK pathways

Progranulin inhibits LPS-induced macrophage M1 polarization via NF-кB and MAPK pathways

  • BMC Immunol. 2020 Jun 5;21(1):32. doi: 10.1186/s12865-020-00355-y.
Lianlian Liu 1 2 3,  Hongmei Guo 1,  Aimei Song 1 2 3,  Jiahui Huang 1,  Yu Zhang 1,  Shanshan Jin 1,  Shutong Li 1,  Liguo Zhang 1,  Chengzhe Yang 4,  Pishan Yang 5 6 7
Affiliations

Affiliations

  • 1 Department of Periodontology, School of Stomatology, Shandong University, 44 West Wenhua Road, Jinan, 250012, Shandong, People's Republic of China.
  • 2 Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan, Shandong, China.
  • 3 Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, Shandong, China.
  • 4 Department of Oral and Maxillofacial Surgery, Qilu Hospital and Institute of Stomatology, Shandong University, Jinan, 250012, Shandong, People's Republic of China. [email protected].
  • 5 Department of Periodontology, School of Stomatology, Shandong University, 44 West Wenhua Road, Jinan, 250012, Shandong, People's Republic of China. [email protected].
  • 6 Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan, Shandong, China. [email protected].
  • 7 Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, Shandong, China. [email protected].
PMID: 32503416 DOI: 10.1186/s12865-020-00355-y
Abstract

Background: Macrophage M1 polarization plays a pivotal role in inflammatory diseases. Progranulin (PGRN) has potential anti-inflammation action, however, the effect of PGRN on macrophage M1 polarization has been poorly studied. Our study aimed to investigate the effect of PGRN on lipopolysaccharide (LPS)-induced macrophage M1 polarization and clarify the underlying mechanisms.

Methods: RAW264.7 cells were polarized to M1 macrophage by LPS with or without recombinant PGRN (rPGRN) and tumor necrosis factor alpha antibody (anti-TNF-α). A cell counting kit-8 assay (CCK-8), flow cytometry, Quantitative Real-Time PCR assay (q-PCR), Western blot assay and enzyme-linked immunosorbent assay (ELISA) were used to determine the effect of different treatments on cell proliferation, expression of surface phenotype marker and expressions and secretion of inflammatory cytokines. The activation of NF-κB/mitogen-activated protein kinase (MAPK) pathways and the nuclear translocation of NF-κB p65 were detected by Western blot and immunofluorescence respectively. THP-1 and primary bone marrow-derived monocytes (BMDMs) were also used to demonstrate effect of PGRN on expressions and secretion of inflammatory cytokines induced by LPS.

Results: In RAW264.7 cells, rPGRN at concentrations below 80 ng/ml significantly promoted cell proliferation in dose dependent fashion. rPGRN significantly inhibited LPS-induced change of phenotype (CD86/CD206 ratio) and function (tumor necrosis factor (TNF-α) and inducible nitric oxide synthase (iNOS) expressions). LPS-stimulated secretion of TNF-α and activated phosphorylation of IKKα/β, IкBα, p65, JNK and p38 and the nucleus translocation of NF-кB p65 were also significantly downregulated by rPGRN. In addition, recombinant TNF-α (rTNF-α) significantly boosted TNF-α and iNOS expression vs the control group. Moreover, anti-TNF-α significantly inhibited LPS-induced TNF-α and iNOS expression. In THP-1 and BMDM cells, reversing effect of rPGRN on LPS-enhanced expressions of TNF-α and iNOS and secretion of TNF-α was further demonstrated.

Conclusions: PGRN down-regulates LPS-induced macrophage M1 polarization in phenotype and function via NF-κB/MAPK signaling pathways.

Keywords

Lipopolysaccharide (LPS); MAPK; Macrophage polarization; NF-κB; PGRN.

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