A new tumor necrosis factor（TNF）-α regulator,lipopolysaccharides-induced TNF-α factor,is associated with obesity and insulin resistance

Background Tumor necrosis factor （TNF）-α plays an important role in mediating inflammatory state in obesity and related disorders.Lipopolysaccharides （LPS）-induced TNF-α factor （LITAF） is recently verified as a regulator of TNF-α and other inflammatory cytokines,and maybe act as a transcriptional factor.The aim of this study was to confirm the association between LITAF and obesity and insulin resistance.Methods Forty-seven subjects with a wide range of body mass index （BMI） were included.Subjects were divided intothree groups according to the criteria of normal weight,overweight and obese.Anthropometrics and metabolic profile were tested for all the subjects.Peripheral monocytes were isolated and purified.LITAF transcription was detected by real time PCR,and the protein expression in whole cell and nucleus extracts was detected by Western blotting analysis;transcriptional activity of LITAF was detected by ELISA like assay using a probe containing the DNA binding sequence of LITAF.Plasma TNF-α and interleukin （IL）-6 concentrations were determined with ELISA kit.Results The LITAF mRNA and protein expression in whole cell were higher in overweight （P 〈0.05） and obese group （P 〈0.05） compared with that in normal weight group.The LITAF protein expression in the nucleus and transcriptional activity could not be detected.LITAF protein expression was positively correlated with BMI （r=0.541,P 〈0.001）,waist circumference （r=0.391,P=0.007）,the homeostasis model assessment for insulin resistance （r=0.372,P=0.011） and fasting insulin levels （r=0.359,P=0.013）.As a regulator of inflammatory cytokines,LITAF protein expression was positively correlated with plasma TNF-α （r=0.621,P=0.002） and IL-6 （r=0.407,P=0.039） concentration.Multiple variant regression analysis indicated that BMI （P=0.002） and waist circumference （P=0.017） were independent predictors of LITAF protein expression.Conclusions LITAF is associated with obesity and insulin resistance,as well as inflammatory cytokine secretion.The results indicate LITAF to be a new mediator between inflammation and the obesity related disorders.

Anti-cancer and anti-inflammatory activities of aronia (Aronia melanocarpa) leaves

Objective: To determine the anti-cancer effect of aronia leaf extract on SK-Hep1 cells using migration, metallo metrix proteinase-2/-9(MMP-2/-9) and MT-1 MMP expression and to evaluate the anti-inflammatory activities of the leaf extract. Methods: The effect of aronia leaf extract on cancer prevention was investigated. SK-Hep1 human liver cancer cell line was treated with aronia leaf extract at various concentractions. MTT assay was used to measure cancer cell growth inhibition, and wound migration assay was used for metastasis determination. The expression of MMP-2/-9 was measured at the protein level using zymography and the expression of MMP-2/-9 and MT-1 MMP was examined at the gene level by RT-PCR. Raw 264.7 macrophage cells were stimulated with lipopolysaccharides to induce inflammation, and then the inhibition of inflammation was evaluated by treatment of aronia leaf extract. Expressions of interleukin-6, tumor factor-d. Results: SK-Hep1 cell growth was inhibited毩, and nitric oxide(NO) were also determine in proportion to the concentration of aronia leaf extract. In migration assay, aronia leaf extract showed 61.3%-96.3% wound size inhibtion after treating 50-200 μg/mL of aronia leaf extract for 24 h. At the protein level, the expression of MMP-2 and MMP-9 decreased as the concentration of aronia leaf extract treated with SK-Hep1 cells increased. In addition, the same pattern as in the protein was also observed in the mRNA levels. The expressions of MMP-2 and MMP-9 protein were inhibited by 92.2% and 53.8%, respectively after treatment with 200 μg/mL aronia leaf extract. In addition, Raw 264.7 cells treated with aronia leaf extract did not affect cell survival. There was dose dependent inhibition of interleukine-6, tumor necrosis factor-a and nitric oxide after treating aronia leaf extract in lipopolysaccharides-treated Raw cell. Conclusions: The results show that aronia leaf has anticancer and and antimetastatic properties in SK-Hep1 and Raw 264.7 cells.