Advanced application of porcine intramuscular adipocytes for evaluating anti-adipogenic and anti-inflammatory activities of immunobiotics

We previously established a clonal porcine intramuscular preadipocyte (PIP) line and we were able to establish a protocol to obtain functionalmature adipocytes fromPIP cells. We hypothesized that both PIP cells and mature adipocytes are likely to be useful in vitro tools for increasing our understanding of immunobiology of adipose tissue, and for the selection and study of immunoregulatory probiotics (immunobiotics) able to modulate adipocytes immune responses. In this study, we investigated the immunobiology of PIP cells and mature adipocytes in relation to their response to TNF-α stimulation. In addition, we evaluated the possibility that immunobiotic microorganisms modify adipogenesis and immune functions of porcine adipose tissue through Peyer?s patches (PPs) immune-competent cells.We treated the porcine PPs immune cells with different probiotic strains; and we evaluated the effect of conditioned media from probiotic-stimulated immune cells in PIP cells and mature adipocytes. The Lactobacillus GG and L. gasseri TMC0356 showed remarkable effects, and were able to significantly reduce the expression of pro-inflammatory factors and negative regulators (A20,Bcl-3, and MKP-1) in adipocytes challenged with TNF-α. The results of this study demonstrated that the evaluation of IL-6, and MCP-1 production, and A20 and Bcl-3 down-regulation in TNF-α-challenged adipocytes could function as biomarkers to screen and select potential immunobiotic strains. Taking into consideration that several in vivo and in vitro studies clearly demonstrated the beneficial effects of Lactobacillus GG and L. gasseri TMC0356 in adipose inflammation, the results presented in this work indicate that the PIP cells and porcine adipocytes could be used for the screening and the selection of new immunobiotic strains with the potential to functionally modulate adipose inflammation when orally administered.Fil: Suzuki, Masahiko . Tohoku University. Graduate School of Agricultural Science. Food and Feed Immunology Group; JapónFil: Tada, Asuka . Tohoku University. Graduate School of Agricultural Science. Food and Feed Immunology Group; JapónFil: Kanmani, Paulraj . Tohoku University. Graduate School of Agricultural Science. Food and Feed Immunology Group; JapónFil: Watanabe, Hitoshi. Tohoku University. Graduate School of Agricultural Science. Cell Biology Laboratory; JapónFil: Aso, Hisashi. Tohoku University. Graduate School of Agricultural Science. Cell Biology Laboratory; JapónFil: Suda, Yoshihito . Miyagi University. Department of Food, Agriculture and Environment; JapónFil: Nochi, Tomonori . Tohoku University. Graduate School of Agricultural Science. Cell Biology Laboratory; JapónFil: Miyazawa, Kenji . Takanashi Milk Products Co., Ltd. Technical Research Laboratory; JapónFil: Yoda, Kazutoyo . Takanashi Milk Products Co., Ltd. Technical Research Laboratory; JapónFil: He, Fang . Takanashi Milk Products Co., Ltd. Technical Research Laboratory; JapónFil: Hosoda, Masataka . Takanashi Milk Products Co., Ltd. Technical Research Laboratory; JapónFil: Saito, Tadao . Tohoku University. Graduate School of Agricultural Science. Food and Feed Immunology Group; JapónFil: Villena, Julio Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Centro de Referencia para Lactobacilos (i); ArgentinaFil: Kitazawa, Haruki . Tohoku University. Graduate School of Agricultural Science. Food and Feed Immunology Group; Japó

Induction of TLRs expression in PIP cells and differentiated adipocytes.

<p>Both PIP cells and differentiated adipocytes were pre-stimulated with CFS of various LABs for 48 h and then post-stimulated with TNF-α for 3, 6, and 12 hours. The expressions of TLR2 and TLR4 were evaluated. Cells treated only with TNF-α were used as TNF-α controls. Three or four independent experiments were conducted for each case and the average values (mean ± S.D) were represented as immunomodulatory effect of LABs. The presence of stars (*) indicates statistical differences with significant levels of P<0.05.</p