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Inhibition of Porcine Preadipocytes Differentiation and Reduction of Glucose Induction by Txnip-overexpression |
YU Qi, YUE Xiao-Jing, ZHANG Guo-Hua, DAI Hong-Wei, CHEN Yan, LI Jian-Jiang, LU Jian-Xiong* |
College of Life Science and Engineering, Northwest Minzu University, Lanzhou 730030, China |
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Abstract Thioredoxin interacting protein (Txnip), a redox regulatory protein, binds to thioredoxin (Trx) and inhibits its activity, regulating the redox state in various physiological processes of cells, but its role in differentiation of porcine fat cells regulated by nutrient is unclear. In this study, the cDNA sequence of Sus scrofa Txnip gene was synthesized and the recombinant lentivirus (LV) overexpressing Txnip (LV5-Txnip) was constructed. The isolated porcine preadipocytes were cultured and transfected with LV5-Txnip and the transfection efficiency was around 90%. The Txnip mRNA level was raised by about 80 folds. The porcine preadipocytes transfected with LV5-Txnip were induced by adipogenic differentiation medium and treated with 5 or 15 mmol/L concentration of glucose, respectively. The differentiation and mRNA expressions of adipogenic-related genes for the cells were detected every other day using oil red O staining extraction assay and qRT-PCR, respectively. The results showed that differentiation of the cells transfected with LV5-Txnip was significantly inhibited (P<0.05), compared to the cells transfected with negative control virus and to the non-transfected cells, the expression level of peroxisome proliferator-activated receptor γ (PPARγ) and fatty acid synthase (FAS) was also reduced significantly (P<0.05). The differentiation and mRNA expression of PPARγ, carbohydrate response element binding protein (ChREBP) and glucose transporter 4 (Glut4) of control preadipocytes were induced significantly by high concentration of glucose, but this promotion was reduced by Txnip-overexpression (P<0.05). Txnip-overexpression suppressed differentiation of porcine preadipocytes through down-regulating PPARγ expression, which indicating Txnip could be an inhibitor of porcine adipocyte differentiation. Txnip impaired the promotion of glucose on the cell differentiation via decreased Glut4 expression leading to lower glucose transport and expression of ChREBP and PPARγ. The present study provides a reference for the investigation of the adipogenesis regulated by nutrient.
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Received: 13 January 2020
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Corresponding Authors:
*lu2003jx@163.com
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