Nutr Res Pract.  2016 Dec;10(6):623-628. 10.4162/nrp.2016.10.6.623.

Induction of heme oxygenase-1 with dietary quercetin reduces obesity-induced hepatic inflammation through macrophage phenotype switching

• Kim CS
• Choi HS
• Joe Y
• Chung HT
• Yu R

• KMID: 2359619
• DOI: http://doi.org/10.4162/nrp.2016.10.6.623

Abstract

BACKGROUND/OBJECTIVES
Obesity-induced steatohepatitis accompanied by activated hepatic macrophages/Kupffer cells facilitates the progression of hepatic fibrinogenesis and exacerbates metabolic derangements such as insulin resistance. Heme oxyganase-1 (HO-1) modulates tissue macrophage phenotypes and thus is implicated in protection against inflammatory diseases. Here, we show that the flavonoid quercetin reduces obesity-induced hepatic inflammation by inducing HO-1, which promotes hepatic macrophage polarization in favor of the M2 phenotype.
MATERIALS/METHODS
Male C57BL/6 mice were fed a regular diet (RD), high-fat diet (HFD), or HFD supplemented with quercetin (HF+Que, 0.5g/kg diet) for nine weeks. Inflammatory cytokines and macrophage markers were measured by ELISA and RT-PCR, respectively. HO-1 protein was measured by Western blotting.
RESULTS
Quercetin supplementation decreased levels of inflammatory cytokines (TNFα, IL-6) and increased that of the anti-inflammatory cytokine (IL-10) in the livers of HFD-fed mice. This was accompanied by upregulation of M2 macrophage marker genes (Arg-1, Mrc1) and downregulation of M1 macrophage marker genes (TNFα, NOS2). In co-cultures of lipid-laden hepatocytes and macrophages, treatment with quercetin induced HO-1 in the macrophages, markedly suppressed expression of M1 macrophage marker genes, and reduced release of MCP-1. Moreover, these effects of quercetin were blunted by an HO-1 inhibitor and deficiency of nuclear factor E2-related factor 2 (Nrf2) in macrophages.
CONCLUSIONS
Quercetin reduces obesity-induced hepatic inflammation by promoting macrophage phenotype switching. The beneficial effect of quercetin is associated with Nrf2-mediated HO-1 induction. Quercetin may be a useful dietary factor for protecting against obesity-induced steatohepatitis.

Keyword

Obesity; quercetin; inflammation; macrophage

MeSH Terms

Animals
Blotting, Western
Coculture Techniques
Cytokines
Diet
Diet, High-Fat
Down-Regulation
Enzyme-Linked Immunosorbent Assay
Fatty Liver
Heme Oxygenase-1*
Heme*
Hepatocytes
Humans
Inflammation*
Insulin Resistance
Liver
Macrophages*
Male
Mice
NF-E2-Related Factor 2
Obesity
Phenotype*
Quercetin*
Up-Regulation
Cytokines
Heme
Heme Oxygenase-1
NF-E2-Related Factor 2
Quercetin

Figure

• 

  Fig. 1 Effects of quercetin on hepatic inflammatory responses and macrophage phenotypes in HFD-fed obese mice. (A) Inflammatory cytokines (TNFα, IL-6, and MCP-1) and (B) an anti-inflammatory cytokine (IL-10) in liver were determined as described in Methods. Results are means ± SEM of six mice per group. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with obese mice fed an HFD. (C) F4/80 as a macrophage marker and (D-E) M1 and M2 macrophages mRNAs in livers were determined by real-time PCR. The intensity of the bands was densitometrically measured and normalized to GAPDH transcripts. Results are means ± SEM of six mice per group. * P < 0.05, ** P < 0.01 compared with obese mice fed an HFD.

• 

  Fig. 2 Effect of HO-1 inhibition on quercetin action in co-cultures of lipid-laden hepatocytes with macrophages. Primary hepatocytes were treated with palmitic acid (BSA: palmitic acid ratio 1:5) for 12h. Raw 264.7 macrophages were then co-cultured with the lipid-laden hepatocytes for 12h in the presence or absence of quercetin or ZnPP. L-Hepa; lipid-laden hepatocytes, Co-culture; lipid-laden hepatocytes and macrophages, Que; quercetin. (A) HO-1 protein was measured by western blotting in macrophages. Results are means ± SEM of two experiments with duplicate determinations. * P < 0.05, *** P < 0.001 compared with no treatment. (B) MCP-1 production detected by ELISA in co-cultured lipid-laden hepatocytes and macrophages. Results are means ± SEM of triplicate samples. * P < 0.05, *** P < 0.001 compared with quercetin treatment. (C) M1 macrophage mRNAs in co-cultured lipid-laden hepatocytes and macrophages determined by real-time PCR. Results are means ± SEM of quadruplicate samples. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with quercetin treatment.

• 

  Fig. 3 Effect of Nrf2 deficiency on quercetin action in LPS-stimulated macrophages. (A) HO-1 protein was measured by western blotting in peritoneal macrophages of Nrf2 KO and WT mice. Results are means ± SEM of three independent experiments. ** P < 0.01 compared with LPS treatment. (B-C) M1/M2 marker mRNAs were determined by real-time PCR. The intensity of the bands was densitometrically measured and normalized to GAPDH transcripts. Results are means ± SEM of two experiments with duplicate determinations. * P < 0.05, ** P < 0.01, *** P < 0.001 compared with LPS treatment.

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