Evaluation of Fat Accumulation and Adipokine Production during the Long-Term Adipogenic Differentiation of Porcine Intramuscular Preadipocytes and Study of the Influence of Immunobiotics

Leptin 0301 basic medicine FAT ACCUMULATION Swine Epidemiology Physiology Beige Adipocytes long-term adipogenic differentiation (LTAD) Cell Count Brown Adipose Tissue Function and Physiology ADIPOKINES Ligands IMMUNOBIOTICS Endocrinology https://purl.org/becyt/ford/3.1 Adipocytes Andrology adipokines Internal medicine Adiposity 2. Zero hunger 0303 health sciences Adipogenesis Muscles Fatty Acids Toll-Like Receptors Rehabilitation 3. Good health Chemistry Adipose Tissue PORCINE INTRAMUSCULAR ADIPOCYTE porcine intramuscular adipocyte Medicine Adiponectin Adipose tissue Article Cell Line 03 medical and health sciences INFLAMMATION Adipokines Adipokine Health Sciences Animals The Role of Exercise in Inflammation and Oxidative Stress https://purl.org/becyt/ford/3 Obesity Biology Cell Proliferation Cell Size Inflammation QH573-671 long-term adipogenic differentiation (LTAD), inflammation Tumor Necrosis Factor-alpha Insulin resistance immunobiotics LONG-TERM ADIPOGENIC DIFFERENTIATION (LTAD) fat accumulation Adaptive Thermogenesis Inflammation and Obesity-Related Metabolic Disorders inflammation FOS: Biological sciences Stimulation Cytology
DOI: 10.3390/cells9071715 Publication Date: 2020-07-21T10:38:55Z
ABSTRACT
The degree of fat accumulation and adipokine production are two major indicators of obesity that are correlated with increased adipose tissue mass and chronic inflammatory responses. Adipocytes have been considered effector cells for the inflammatory responses due to their capacity to express Toll-like receptors (TLRs). In this study, we evaluated the degree of fat accumulation and adipokine production in porcine intramuscular preadipocyte (PIP) cells maintained for in vitro differentiation over a long period without or with stimulation of either TNF-α or TLR2-, TLR3-, or TLR4-ligands. The cytosolic fat accumulation was measured by liquid chromatography and the expression of adipokines (CCL2, IL-6, IL-8 and IL-10) were quantified by RT-qPCR and ELISA at several time points (0 to 20 days) of PIP cells differentiation. Long-term adipogenic differentiation (LTAD) induced a progressive fat accumulation in the adipocytes over time. Activation of TLR3 and TLR4 resulted in an increased rate of fat accumulation into the adipocytes over the LTAD. The production of CCL2, IL-8 and IL-6 were significantly increased in unstimulated adipocytes during the LTAD, while IL-10 expression remained stable over the studied period. An increasing trend of adiponectin and leptin production was also observed during the LTAD. On the other hand, the stimulation of adipocytes with TLRs agonists or TNF-α resulted in an increasing trend of CCL2, IL-6 and IL-8 production while IL-10 remained stable in all four treatments during the LTAD. We also examined the influences of several immunoregulatory probiotic strains (immunobiotics) on the modulation of the fat accumulation and adipokine production using supernatants of immunobiotic-treated intestinal immune cells and the LTAD of PIP cells. Immunobiotics have shown a strain-specific ability to modulate the fat accumulation and adipokine production, and differentiation of adipocytes. Here, we expanded the utility and potential application of our in vitro PIP cells model by evaluating an LTAD period (20 days) in order to elucidate further insights of chronic inflammatory pathobiology of adipocytes associated with obesity as well as to explore the prospects of immunomodulatory intervention for obesity such as immunobiotics.
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