The inhibition of inflammatory molecule expression on 3T3-L1 adipocytes by berberine is not mediated by leptin signaling

In our previous study, we have shown that berberine has both anti-adipogenic and anti-inflammatory effects on 3T3-L1 adipocytes, and the anti-adipogenic effect is due to the down-regulation of adipogenic enzymes and transcription factors. Here we focused more on anti-inflammatory effect of berberine using real time RT-PCR and found it changes expressions of adipokines. We hypothesized that anti-adipogenicity of berberine mediates anti-inflammtory effect and explored leptin as a candidate mediator of this signaling. We studied this hypothesis by western blot analysis, but our results showed that berberine has no effect on the phosphorylations of STAT-3 and ERK which have important roles on leptin signaling. These results led us to conclude that the anti-inflammatory effect of berberine is not mediated by the inhibition of leptin signal transduction. Moreover, we have found that berberine down-regulates NF-kappaB signaling, one of the inflammation-related signaling pathway, through western blot analysis. Taken together, the anti-inflammatory effect of berberine is not mediated by leptin, and berberine induces anti-inflammatory effect independent of leptin signaling.

Berberine reduces the expression of adipogenic enzymes and inflammatory molecules of 3T3-L1 adipocyte

Berberine (BBR), an isoquinoline alkaloid, has a wide range of pharmacological effects, yet its exact mechanism is unknown. In order to understand the anti-adipogenic effect of BBR, we studied the change of expression of several adipogenic enzymes of 3T3-L1 cells by BBR treatment. First, we measured the change of leptin and glycerol in the medium of 3T3-L1 cells treated with 1 micrometer, 5 micrometer and 10 micrometer concentrations of BBR. We also measured the changes of adipogenic and lipolytic factors of 3T3-L1. In 3T3-L1 cells, both leptin and adipogenic factors (SREBP-1c, C/EBP-alpha, PPAR-gamma, fatty acid synthase, acetyl-CoA carboxylase, acyl-CoA synthase and lipoprotein lipase) were reduced by BBR treatment. Glycerol secretion was increased, whereas expression of lipolytic enzymes (hormone-sensitive lipase and perilipin) mRNA was slightly decreased. Next, we measured the change of inflammation markers of 3T3-L1 cells by BBR treatment. This resulted in the down-regulation of mRNA level of inflammation markers such as TNF-alpha, IL-6, C- reactive protein and haptoglobin. Taken together, our data shows that BBR has both anti-adipogenic and anti-inflammatory effects on 3T3-L1 adipocytes, and the anti-adipogenic effect seems to be due to the down-regulation of adipogenic enzymes and transcription factors.

Active Immunization Study of Colon Cancer Derived 1-8D Peptide in HHD Mice

BACKGROUND: 1-8D gene is a member of human 1-8 interferon inducible gene family and was shown to be overexpressed in fresh colon cancer tissues. Three peptides 1-6, 3-5 and 3-7 derived from human 1-8D gene were shown to have immunogenicity against colon cancer. METHODS: To study tumor immunotherapy of three peptides we established an active immunization model using HHD mice. D(b-/-) x beta2 microglobulin (beta2 m) null mice transgenic for a chimeric HLA-A2.1/D(b-)beta2 m single chain (HHD mice) were challenged with B16/HHD/1-8D tumor cells and were immunized with irradiated peptide-loaded RMA- S/HHD/B7.1 transfectants. In therapy model tumor growth was retarded in HHD mice that were injected with 3-5 peptide-loaded RMA-S/HHD/B7.1. In survival test vaccination with 1-8D-derived peptide protects HHD mice from tumor progression after tumor challenge. RESULTS: These studies show that peptide 3-5 derived from 1-8D gene can be the most effective candidate for the vaccine of immunotherapy against colon cancer and highlight 1-8D gene as putative colon carcinoma associated antigens. CONCLUSION: We demonstrated that RMA-S/HHD/ B7.1 loaded with 1-8D peptides, especially 3-5, immunization generates potent antitumor immunity against tumor cells in HHD mice and designed active immunization as proper immunotherapeutic protocols.

Adoptive Transfer of Colon Cancer Derived Peptide-specific CD8+ T Cells in HHD Mice

BACKGROUND: 1-8D gene is a member of human 1-8 interferon inducible gene family and is shown to be overexpressed in fresh colon cancer tissues. Three peptides 1-6, 3-5 and 3-7 derived from 1-8D gene were shown to have immunogenicity against colon cancer. METHODS: To study tumor immunotherapy of these peptides we established an adoptive transfer model. D(b-/-)Xbeta2 microglobulin (beta2m) null mice transgenic for a chimeric HLA-A2.1/D(b)-beta2m single chain (HHD mice) were immunized with irradiated peptide-loaded RMA-S/HHD/B7.1 transfectants. Spleens were removed after last immunization, and splenocytes were re-stimulated in vitro. Lymphocytes from vaccinated HHD mice were transferred together with IL-2 to the tumor bearing nude mice that were challenged S.C. with the HCT/HHD/B7 colon carcinoma cell line that was found to grow in these mice. RESULTS: Peptide 3-5 was found to be highly effective in CTL activity. Adoptively transferred anti-peptide 3-5 cytolytic T lymphocytes caused significant retardation in tumor growth. CONCLUSION: This study shows that peptide 3-5 can be the most effective candidate for the vaccine of adoptive immunotherapy against colon cancer.

The Standardization of the Ge riatric Quality of Life s cale-Dementia(GQOL-D)

Objectives : We developed the instrument to assess the quality of life(QOL) for demented patients, 'Geriatric Quality of Life-Dementia(GQOL-D)'. The purpose of this study was to standardize the GQOL-D and to introduce administration results in Korean demented patients. METHODS: The normal subjects were 340 elderly people, aged over 55 years old, who were physically and cognitively normal. And the patient group was consisted of 69 demented patients. RESULTS: Both in normal group and in patient group, the internal consistency, Cronbach's = 0.87, 0.90, respectively, and item-total correlation was acceptable. And the test-retest reliability revealed the stability across time by r=0.86, 0.77, respectively. Criterion validity was found to be a high correlation between each itemand overall QOL' item, and adequate correlations between the GQOL-D and scales assessing cognition, psychological wellbeing, behavior and activities of daily living convinced convergent validity. In the result of factor analysis, 2 factors with a variance percentage of 37.7% were extracted. The mean score and standard deviation of items of the GQOL-D for two groups and differences in items such as memory, recreation/leisure, self esteem, general health, and mobility between groups were presented. CONCLUSION: It was confirmed that there is no difficulty for old people to carry out the GQOL-D. As it was suggested that there is enough room for intervening in dementia patients' emotional and behavioral difficulties, the GQOL-D will be useful in research and clinical practice for the patients with dementia.