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. 2023 Mar 17;24(6):5745.
doi: 10.3390/ijms24065745.

Asprosin Enhances Cytokine Production by a Co-Culture of Fully Differentiated Mature Adipocytes and Macrophages Leading to the Exacerbation of the Condition Typical of Obesity-Related Inflammation

Agnieszka I Mazur-Bialy  1
Affiliations

Asprosin Enhances Cytokine Production by a Co-Culture of Fully Differentiated Mature Adipocytes and Macrophages Leading to the Exacerbation of the Condition Typical of Obesity-Related Inflammation

Agnieszka I Mazur-Bialy. Int J Mol Sci. .

Abstract

Asprosin, a fasting-induced, glucogenic, and orexigenic adipokine, has gained popularity in recent years as a potential target in the fight against obesity and its complications. However, the contribution of asprosin to the development of moderate obesity-related inflammation remains still unknown. The present study aimed to evaluate the effect of asprosin on the inflammatory activation of adipocyte-macrophage co-cultures at various stages of differentiation. The study was performed on co-cultures of the murine 3T3L1 adipocyte and the RAW264.7 macrophage cell lines treated with asprosin before, during, and after 3T3L1 cell differentiation, with or without lipopolysaccharide (LPS) stimulation. Cell viability, overall cell activity, and the expression and release of key inflammatory cytokines were analyzed. In the concentration range of 50-100 nM, asprosin increased the pro-inflammatory activity in the mature co-culture and enhanced the expression and release of tumor necrosis factor α (TNF-α), high-mobility group box protein 1 (HMGB1), and interleukin 6 (IL-6). Macrophage migration was also increased, which could be related to the upregulated expression and release of monocyte chemoattractant protein-1 (MCP-1) by the adipocytes. In summary, asprosin exerted a pro-inflammatory effect on the mature adipocyte-macrophage co-culture and may contribute to the spread of moderate obesity-associated inflammation. Nevertheless, further research is needed to fully elucidate this process.

Keywords: adipocytes; asprosin; inflammation; macrophages; obesity; obesity-related inflammation.

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Conflict of interest statement

The author declares no conflict of interest.

Figures

Figure 1
Figure 1
Effects of asprosin on overall cell activity measured in a CellTiter test (A) and viability (B) of adipocyte–macrophage co-cultures after 7, 14, and 21 days of differentiation, with lipopolysaccharide (LPS) or (C) without LPS (100 ng/mL) stimulation. The results from five independent experiments are expressed as mean + standard deviation (SD). Statistical significance (p < 0.05) was determined by the Duncan t-test.
Figure 2
Figure 2
Effects of asprosin on lipopolysaccharide (LPS; 100 ng/mL)-induced cytokine mRNA expression and release: tumor necrosis factor-alpha (TNF-α) (A), interleukin-6 (IL-6) (B), and interleukin-1 beta (IL-1β) (C) in adipocyte–macrophage cocultures (upper panel) and adipocyte and macrophage monocultures (lower panel) for TNF-α (D), IL-6 (E), and IL-1β (F)on the 21st day of differentiation. The results are expressed as mean + standard deviation (SD) from five independent experiments. Statistical significance (p < 0.05) was determined by the Duncan t-test. The asterisk (*) above the bar denotes statistically significant differences in protein or mRNA levels, calculated relative to the A0 + LPS control group. Cells: (M) macrophages; (A) adipocytes.
Figure 3
Figure 3
Effects of asprosin on lipopolysaccharide (LPS; 100 ng/mL)-induced cytokine mRNA expression and release of high-mobility group box-1 protein (HMGB1) in adipocyte “A” and macrophage “M” monocultures (A) and adipocyte–macrophage cocultures (B) on the 21st day of differentiation. The results are expressed as mean + standard deviation (SD) from five independent experiments. Statistical significance (p < 0.05) was determined by the Duncan t-test. The asterisk (*) above the bar denotes statistically significant differences in protein or mRNA levels, calculated relative to the A0 + LPS control group. Cells: (M) macrophages; (A) adipocytes.
Figure 4
Figure 4
Effects of asprosin on LPS (100 ng/mL)-induced leptin expression and release (A) or adiponectin expression and release (B) by adipocyte–macrophage cocultures at 21 days of differentiation and leptin expression and release (C) or adiponectin expression and release (D) by adipocyte monocultures at 21 days of differentiation. 3T3 L1 adipocytes and RAW 264.7 macrophages were cultured in 12-well dishes with inserts (0.4 µm pore size) for 6 h (mRNA expression, rt-PCR) or 24 h (adipokine release, ELISA tests) with or without asprosin (0, 10, 50, or 100 nM; A0, A10, A50, or A100, respectively). The results are expressed as mean + standard deviation (SD) from five independent experiments. Statistical significance (p < 0.05) was determined by the Duncan t-test. The asterisk (*) above the bar denotes statistically significant differences in protein or mRNA levels, calculated relative to the A0 + LPS control group.
Figure 5
Figure 5
Effects of asprosin on (A) the migration of RAW 264.7 macrophages to the conditioned medium collected from co-culture or adipocytes cultured in the presence of various asprosin concentrations (0, 10, 50, or 100 nM) for 24 h (after 21 days of differentiation). Normal medium was used as a negative control (CTR−); N-formylmethionyl-leucyl-phenylalanine (fMLP) was used as a positive control (CTR+). (B) The effects of asprosin on lipopolysaccharide (LPS)-induced monocyte chemotactic protein 1 (MCP-1) mRNA expression and release by adipocyte–macrophage cocultures or adipocytes. The results are expressed as mean + standard deviation (SD) from five independent experiments. Statistical significance (p < 0.05) was determined by the Duncan t-test. The asterisk (*) above the bar denotes statistically significant differences in migration, protein or mRNA levels, calculated relative to the A0 (A) or A0 + LPS (100 ng/mL) control group. Cells: (M) macrophages; (A) adipocytes.
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References

    1. Romere C., Duerrschmid C., Bournat J., Constable P., Jain M., Xia F., Saha P.K., Del Solar M., Zhu B., York B., et al. Asprosin, a Fasting-Induced Glucogenic Protein Hormone. Cell. 2016;165:566–579. doi: 10.1016/j.cell.2016.02.063. - DOI - PMC - PubMed
    1. Duerrschmid C., He Y., Wang C., Li C., Bournat J.C., Romere C., Saha P.K., Lee M.E., Phillips K.J., Jain M., et al. Asprosin is a centrally acting orexigenic hormone. Nat. Med. 2017;23:1444–1453. doi: 10.1038/nm.4432. - DOI - PMC - PubMed
    1. Li E., Shan H., Chen L., Long A., Zhang Y., Liu Y., Jia L., Wei F., Han J., Li T., et al. OLFR734 Mediates Glucose Metabolism as a Receptor of Asprosin. Cell Metab. 2019;30:319–328. doi: 10.1016/j.cmet.2019.05.022. - DOI - PubMed
    1. Wang C.-Y., Lin T.-A., Liu K.-H., Liao C.-H., Liu Y.-Y., Wu V.C.-C., Wen M.-S., Yeh T.-S. Serum asprosin levels and bariatric surgery outcomes in obese adults. Int. J. Obes. 2019;43:1019–1025. doi: 10.1038/s41366-018-0248-1. - DOI - PubMed
    1. Ugur K., Aydin S. Saliva and Blood Asprosin Hormone Concentration Associated with Obesity. Int. J. Endocrinol. 2019;2019:1–8. doi: 10.1155/2019/2521096. - DOI - PMC - PubMed
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