Deficiency of lipopolysaccharide binding protein facilitates adipose browning, glucose uptake and oxygen consumption in mouse embryonic fibroblasts via activating PI3K/Akt/mTOR pathway and inhibiting autophagy

This research aimed look around the role of lipopolysaccharide-binding protein (LBP) in adipose browning. Mouse embryonic fibroblasts (MEFs) were given differentiation induction reagents and Perifosine (Akt inhibitor), using the transfection of Atg5, short hairpin RNA targeting LBP (shLBP), and Atg5 (shAtg5). The expression amounts of LBP, inflammatory markers , brown fat markers, fat metabolic process marker, autophagy markers, insulin signaling-related molecules , p-mTOR, mTOR, p-Akt, Akt, p-PI3K, and PI3K were Perifosine quantified or based on Western blot, qRT-PCR, and immunofluorescence assay. The development of fat was examined through Oil red O staining assay. The intake of oxygen was assessed utilizing a Seahorse XF96 analyzer, and also the uptake of glucose was evaluated by [3H]-2-deoxy-D-glucose uptake assay. Lack of LBP promoted adipose browning, oxygen consumption, glucose uptake, and insulin sensitivity in differentiated MEFs, where it inhibited inflammation and autophagy. All the effects above were reversed by Atg5 overexpression. Meanwhile, the knockdown of Atg5 strengthened the activation of PI3K/Akt/mTOR path caused through the depletion of LBP, while Perifosine partially reversed the activation of differentiated MEFs. The knockdown of LBP facilitated adipose browning, glucose uptake, and oxygen consumption in MEFs through the activation of PI3K/Akt/mTOR path and also the inhibition of autophagy.