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Lipopolysaccharide-induced cytokine expression pattern in peripheral blood mononuclear cells in childhood obesity.

Molecular medicine reports (2016-10-26)
Yu Wu, Bin Yue, Jia Liu
ABSTRAKT

Obesity is characterized by the abnormal or excessive fat accumulation that may impair health and extensive increase in body mass index (BMI). Childhood obesity may occur due to disturbances in metabolic regulation, which lead to metabolic syndrome and other diseases. Peripheral blood suspended immune cells are responsible for immune surveillance. The aim of the present study was to map the inflammatory cytokine expression pattern of isolated peripheral blood mononuclear cells (PBMCs) following lipopolysaccharide (LPS) stress in vitro and clinical chemistry parameters in the plasma of subjects. PBMCs were isolated through density gradient ultracentrifugation of the blood from obese infants that would reflect the cytokine response. Isolated PBMCs were cultured in vitro in RPMI‑1640 medium and stressed with 1 µg LPS in order to investigate the expression pattern of cytokines, such as interleukin (IL)‑2, interferon (IFN)‑γ, IFN‑α, tumor necrosis factor‑α and IL‑6 using enzyme‑linked immunosorbent assays and reverse transcription‑quantitative polymerase chain reaction. Levels of NO, lipid levels, total protein, albumin, marker enzymes aspartate transaminase and alanine transaminase, malondialdehyde (MDA), and reduced glutathione in the plasma were detected. Reduction in the expression of the inflammatory cytokines in PBMCs, reduced protein and globulins, and altered MDA and GSH levels in the plasma were observed. Altered or compromised pro‑inflammatory signals from PBMCs in vitro and the clinical chemistry parameters of the plasma suggested that there were compromised immunological responses in obese children compared with matched controls.

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Sigma-Aldrich
AST Activity Assay Kit, sufficient for 100 colorimetric tests
Sigma-Aldrich
ALT Activity Assay, sufficient for 100 colorimetric or fluorometric tests