3T3-L1 adipocytes express the B-cell-activating factor (BAFF) and three different BAFF receptors (BAFF-Rs). Furthermore, BAFF expression is regulated by inflammatory modulators, such as tumor necrosis factor-alpha and rosiglitazone. Here we investigated the function of BAFF in 3T3-L1 adipocytes and RAW 264.7 macrophages. We examined adipokine expression in 3T3-L1 adipocytes treated with 10 ng ml-1 BAFF. We also examined inflammatory molecule expression in RAW 264.7 macrophages treated with 10 or 100 ng ml-1 BAFF. We examined BAFF expression in the coculture of 3T3-L1 adipocytes and RAW 264.7 macrophages, as well as in white adipose tissue (WAT) of diet-induced obese (DIO) mice. We found that BAFF decreases leptin and adiponectin expression, but increases the expression of proinflammatory adipokines monocyte chemotactic protein-1, interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and haptoglobin. Coculturing the two cell types resulted in increased BAFF mRNA and protein expression, as well as modulation of BAFF-R mRNA expression in both cell types. These data indicate that BAFF might mediate adipocyte and macrophage interaction. When RAW 264.7 macrophages were treated with BAFF, BAFF-R expression was modulated as in coculture, and nitric oxide synthase and IL-6 expression increased. BAFF expression also increased in WAT of DIO mice. We propose that BAFF can regulate adipokine expression and possibly mediate adipocyte and macrophage interaction.
3T3-L1 Cells ; Adipocytes/drug effects/*metabolism ; Adipokines/genetics/*metabolism ; Adiponectin/genetics/metabolism ; Animals ; B-Cell Activating Factor/*metabolism/pharmacology ; Chemokine CCL2/genetics/metabolism ; Coculture Techniques ; Gene Expression Regulation/drug effects ; Haptoglobins/genetics/metabolism ; Inflammation Mediators/metabolism ; Interleukin-6/genetics/metabolism ; Leptin/genetics/metabolism ; Macrophages/drug effects/*metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Obese ; RNA, Messenger/genetics/metabolism

OBJECTIVETo explore the expression of YKL-40, TLR4 and NF-κB in chronic rhinosinusitis (CRS) with or without nasal polyps (CRSwNP and CRSsNP), and to investigate their expressional correlation and the potential role in pathogenesis of CRS.

METHODSThe specimens were obtained from sinus mucosa and inferior turbinate mucosa of the patients with informed consent. The different expression of YKL-40, TLR4 and NF-κB among each group was detected by real time RT-PCR and immunohistochemistry (S-P method). SPSS 17.0 software was used to analyze the data.

RESULTSmRNA level: The relative expression of YKL-40 in CRSwNP group (0.91±0.17) was higher than those in the control group (0.49±0.09), the difference was significant (t=2.12, P<0.05). The relative expression of TLR4 in CRSsNP group (0.88±0.19) and CRSwNP group (0.67±0.13) were lower than those in control group (1.48±0.14), the differences were significant (t value was -4.11, -2.48, all P<0.05). The relative expression of NF-κB in CRSsNP group (0.69±0.13) and CRSwNP group (0.72±0.14) were lower than those in control group (1.20±0.15), the differences were significant (t value was 2.33, 2.27, all P<0.05). Protein level: The expression of YKL-40 in CRSwNP group was stronger than that in CRSsNP group and control group (U value was 72.5 and 73, all P<0.01). The expression of TLR4 in CRSwNP group and CRSsNP group was weaker than that in control group (U value was 62 and 38, all P<0.01). There was a negative correlation between YKL-40 and TLR4 (rmRNA=-0.741, P<0.01; rprotein=-0.46, P<0.05) in CRSwNP group.

CONCLUSIONSThe expression of YKL-40 in pantients with CRSwNP is higher than those in healthy control and CRSsNP patients. There was a negative correlation between YKL-40 and TLR4. Both of them may be involved in the pathogenesis of CRSwNP.

Adipokines ; genetics ; metabolism ; Chitinase-3-Like Protein 1 ; Chronic Disease ; Humans ; Immunohistochemistry ; Lectins ; genetics ; metabolism ; NF-kappa B ; Nasal Mucosa ; Nasal Polyps ; RNA, Messenger ; Rhinitis ; metabolism ; Sinusitis ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Turbinates

Adipose tissue hypoxia has been recognized as the initiation of insulin resistance syndromes. The aim of the present study was to investigate the effects of mangiferin on the insulin signaling pathway and explore whether mangiferin could ameliorate insulin resistance caused by hypoxia in adipose tissue. Differentiated 3T3-L1 adipocytes were incubated under normal and hypoxic conditions, respectively. Protein expressions were analyzed by Western blotting. Inflammatory cytokines and HIF-1-dependent genes were tested by ELISA and q-PCR, respectively. The glucose uptake was detected by fluorescence microscopy. HIF-1α was abundantly expressed during 8 h of hypoxic incubation. Inflammatory reaction was activated by up-regulated NF-κB phosphorylation and released cytokines like IL-6 and TNF-α. Glucose uptake was inhibited and insulin signaling pathway was damaged as well. Mangiferin substantially inhibited the expression of HIF-1α. Lactate acid and lipolysis, products released by glycometabolism and lipolysis, were also inhibited. The expression of inflammatory cytokines was significantly reduced and the damaged insulin signaling pathway was restored to proper functional level. The glucose uptake of hypoxic adipocytes was promoted and the dysfunction of adipocytes was relieved. These results showed that mangiferin could not only improve the damaged insulin signaling pathway in hypoxic adipocytes, but also ameliorate inflammatory reaction and insulin resistance caused by hypoxia.
3T3-L1 Cells ; Adipocytes ; drug effects ; immunology ; Adipokines ; genetics ; immunology ; Animals ; Cell Hypoxia ; drug effects ; Glucose ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; immunology ; Insulin ; metabolism ; Insulin Resistance ; Mice ; NF-kappa B ; genetics ; immunology ; Oxygen ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; immunology ; Xanthones ; pharmacology

White fat cells secrete adipokines that induce inflammation and obesity has been reported to be characterized by high serum levels of inflammatory cytokines such as IL-6 and TNF-alpha. Rheumatoid arthritis (RA) is a prototype of inflammatory arthritis, but the relationship between RA and obesity is controversial. We made an obese inflammatory arthritis model: obese collagen-induced arthritis (CIA). C57BL/6 mice were fed a 60-kcal high fat diet (HFD) from the age of 4 weeks and they were immunized twice with type II collagen (CII). After immunization, the obese CIA mice showed higher arthritis index scores and histology scores and a more increased incidence of developing arthritis than did the lean CIA mice. After treatment with CII, mixed lymphocyte reaction also showed CII-specific response more intensely in the obese CIA mice than lean CIA. The anti-CII IgG and anti-CII IgG2a levels in the sera of the obese CIA mice were higher than those of the lean CIA mice. The number of Th17 cells was higher and the IL-17 mRNA expression of the splenocytes in the obese CIA mice was higher than that of the lean CIA mice. Obese CIA mice also showed high IL-17 expression on synovium in immunohistochemistry. Although obesity may not play a pathogenic role in initiating arthritis, it could play an important role in amplifying the inflammation of arthritis through the Th1/Th17 response. The obese CIA murine model will be an important tool when we investigate the effect of several therapeutic target molecules to treat RA.
Adipokines/immunology/metabolism ; Animals ; *Arthritis, Experimental/genetics/immunology/pathology ; Cell Differentiation/genetics/immunology ; *Collagen Type II/administration & dosage/immunology ; Disease Models, Animal ; Gene Expression ; Humans ; Inflammation/chemically induced/*immunology ; Interleukin-17/metabolism ; Interleukin-6/blood ; Joints/immunology/pathology ; Mice ; Mice, Inbred C57BL ; *Obesity/genetics/immunology/pathology ; *Th17 Cells/immunology/metabolism ; Tumor Necrosis Factor-alpha/blood