Conjugated Linoleic Acid Reduces Lipid Accumulation via Down-regulation Expression of Lipogenic Genes and Up-regulation of Apoptotic Genes in Grass Carp (Ctenopharyngodon idella) Adipocyte In Vitro.

The relationship between conjugated linoleic acid (CLA) and lipogenesis has been extensively studied in mammals and some cell lines, but it is relatively rare in fish, and the potential mechanism of action of CLA reducing fat mass remains unclear. The established primary culture model for studying lipogenesis in grass carp (Ctenopharyngodon idella) preadipocytes was used in the present study, and the objective was to explore the effects of CLA on intracellular lipid and TG content, fatty acid composition, and mRNA levels of adipogenesis transcription factors, lipase, and apoptosis genes in grass carp adipocytes in vitro. The results showed that CLA reduced the size of adipocyte and lipid droplet and decreased the content of intracellular lipid and TG, which was accompanied by a significant down-regulation of mRNA abundance in transcriptional regulators including peroxisome proliferator-activated receptor (PPAR) γ, CCAAT/enhancer-binding protein (C/EBP) α, sterol regulatory element-binding protein (SREBP) 1c, lipase genes including fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), lipoprotein lipase (LPL). Meanwhile, it decreased the content of saturated fatty acids (SFAs) and n - 6 polyunsaturated fatty acid (n-6 PUFA) and increased the content of monounsaturated fatty acid (MUFA) and n - 3 polyunsaturated fatty acid (n-3 PUFA) in primary grass carp adipocyte. In addition, CLA induced adipocyte apoptosis through downregulated anti-apoptotic gene B-cell CLL/lymphoma 2 (Bcl-2) mRNA level and up-regulated pro-apoptotic genes tumor necrosis factor-α (TNF-α), Bcl-2-associated X protein (Bax), Caspase-3, and Caspase-9 mRNA level in a dose-dependent manner. These findings suggest that CLA can act on grass carp adipocytes through various pathways, including decreasing adipocyte size, altering fatty acid composition, inhibiting adipocyte differentiation, promoting adipocyte apoptosis, and ultimately decreasing lipid accumulation.

Penicillium marneffei infection within an osteolytic lesion in an HIV-negative patient.

Penicillium marneffei is a thermally dimorphic pathogenic fungus that causes systemic infection similar to disseminated cryptococcosis. P. marneffei is endemic in Southeast Asia, usually infecting HIV-infected individuals; infection of HIV-negative individuals is extremely rare. Here, we describe a disseminated P. marneffei infection within an osteolytic lesion in an HIV-negative patient. A 40-year-old Chinese woman presented with intermittent fever, generalized lymphadenopathy, and a skin rash. Following a sternum biopsy, the patient was diagnosed with P. marneffei infection. An emission computed tomography bone scan revealed the presence of increased radioactivity in the left clavicle and sternum, indicative of an osteolytic lesion. In addition to reporting this very rare case, we also present a brief review of the literature, highlighting the differences in clinical manifestations between HIV-positive and HIV-negative patients infected with P. marneffei as it applies to our case.

[Effects of erythromycin on hydrogen peroxide-induced interleukin-8 synthesis and regulation of glutathione in human bronchial epithelial cells].

OBJECTIVE: To study the effects of erythromycin on Hydrogen peroxide (H2O2)-induced interleukin-8 synthesis and regulation of glutathione in human bronchial epithelial cells. METHODS: Human bronchial epithelial (16HBE) growth curve was recorded by MTT, cells were divided into three groups (1) control (incubation for 24, 36, 48) (2) H2O2 (Pre-incubation for 24, 36, 48 h before adding H2O2 (3) H2O2 + EM (Pre-incubation EM for 24, 36, 48 h before adding H2O2). IL-8 levels were measured in culture supernatants by ELISA, activation of transcription factor NF-kappaB and AP-1 in HBE was evaluated by Electrophoretic mobility shift assay (EMSA). Intracellular GSH and Gamma-GCS concentrations were measured by spectrophotometric assay, gamma-GCS-HS protein were determined by Western blot. RESULTS: Erythromycin (1 microg/ml, 10 microg/ml) and H2O2 (0.01 mM, 0.1 mM) have no effects on cell growth, Preincubation with EM (5 microg/ml) for 36 h and 48 h significantly inhibit H2O2 (0.01 mmol/L) induced increase of IL-8 levels in HBE supernatants, in the mean time decrease the expression of NF-kB and AP-1. Preincubation with EM (5 microg/ml) for 48 h significantly inhibit H2O2 (0.01 mmol/L) induced increase of gamma-GCS levels, gamma-GCS-HS protein expression and AP-1 binding of gamma-GCS-HS promoter in HBE. However GSH and gamma-GCS-HS protein expression in EM + H2O2 group significantly higher than those in control group. CONCLUSION: Erythromycin inhibits oxidant-mediated IL-8 levels through down-regulation of NF-kB and AP-1 binding in HBE, which can further influence the synthesis of GSH and expression gamma-GCS in HBE.

[Cigarette smoking coacervate up-regulates the expression of gamma-glutamylcysteine synthetase in alveolar epithelial cells mediated by activator protein-1].

OBJECTIVE: To investigate the effects of cigarette smoking coacervate (CSC) on the expression and activation of gamma-glutamylcysteine synthetase (GCS), a rate-limitating enzyme in the synthesis of glutathione (reduced form). METHODS: Rat alveolar epithelial cells of the line CCL149 were cultured and exposed to CSC of the concentrations of 10, 1, and 0.1 microg/ml for 1, 4, 8, 12, 24, and 48 hours respectively. RT-PCR was used to detect the mRNA expression of gamma-GCS, and Western blotting was used to detect the protein expression of gamma-GCS. CCL149 cells were transfected with pGL3/gamma-GCS or blank pGL3 plasmid. The luciferase activity was examined Gel retardation assay was used to detect the binding level of activator protein (AP)-1 with the region of the GCLC promoter in CCL-149 cell. RESULTS: The gamma-GCS mRNA expression levels of the CCL149 cells exposed to CSC > 1 microg/ml for 12, 24, and 48 hours were significantly higher than that of the control group (all P < 0.05). The gamma-GCS protein expression levels of the CCL149 cells exposed to CSC > 1 microg/ml for 12, 24, and 48 hours were significantly higher than that of the control group (all P < 0.05). The gamma-GCS protein activity of the CCL149 cells treated with CSC of the concentrations of 10 microg/ml and 1 microg/ml decreased 1, 4, and 8 hours after and then increased in comparison with the control group (all P < 0.05). The gamma-GCS protein activity levels of the CCL149 cells treated with CSC of the concentration of 0.1 microg/ml for less than 48 hours was not significantly different from those of the control group (all P > 0.05), and the gamma-GCS protein activity level of the CCL149 cells treated with CSC of the concentration of 0.1 microg/ml for 48 hours was significantly higher than that of the control group (P < 0.05). The activity of the luciferase with the plasmids containing 5-flanking regulatory region of rat GCLC gene and the activity of gamma-GCS in the CCL149 cells significantly increased after stimulation of CSC for 12, 24 and 48 hours (all P < 0.05). The binding levels of AP-1 with the region of the GCLC promoter in the CCL149 cells treated with CSC for 12, 24, and 48 hours were significantly increased. CONCLUSION: CSC up-regulates the expression of gamma-GCS by activation of the redox-sensitive transcription factor AP-1.