Fig 1: Schematic diagram on the protective role of palmitoleate against inflammation and inflammasome activity. Palmitoleate blocks NF-κB p65 subunit nuclear translocation, thereby blocking LPS-mediated activation of NF-κB pathway and proinflammatory cytokine gene upregulation. Palmitoleate also blocks NLRP3 inflammasome-mediated cleavage of procaspase 1 and pro-IL-1β to active caspase 1 p10/p20 subunits and IL-1β, respectively, thereby preventing LPS and saturated free fatty acid–induced cellular death and inflammation. Further, palmitoleate also prevents LPS + palmitate (PA)-induced cellular MAPK and inflammatory responses. Illustration was made using BioRender. IL, interleukin; LPS, lipopolysaccharide; MAPK, mitogen-activated protein kinase; NLRP3, NACHT, LRR, and PYD domain–containing protein 3.
Fig 2: Palmitoleate (PO) protects against LPS-induced inflammation by blocking NF-κB nuclear translocation or activation. BMDMs were stimulated with LPS (10 ng/ml) alone or in presence of PO (pretreatment for 16 h at 200 μM) for 5–10 min and analyzed for NF-κB signaling pathway activation. Immunoblot analysis showed decreased levels of IκBα after 10 min of LPS exposure (A). Pretreatment of PO however did not block the degradation of IκBα (A). LPS exposure (5–10 min) also increased phosphorylated forms of IKKα/β and PO did not alter the levels of phospho-IKKα/β after 5–10 min. Immunoblot analysis also showed that total IKK-α, IKK-β, and β-actin levels remained unchanged in all the treatment conditions tested (A). We also observed increased phosphorylation of JNK following LPS exposure, and treatment of palmitoleate did not change the levels of phosphorylated JNK (A). Total JNK level remained the same in all experimental conditions (A). NF-κB p65 nuclear translocation were tested by immunoblot following LPS exposure in BMDMs and observed to be increased in the nuclei, suggesting increased nuclear translocation of p65 subunit. Pretreatment of PO for 16 h followed by LPS exposure showed decreased nuclear levels of p65 subunit in BMDMs (B). Histone deacetylase 1 was used as nuclear loading control and showed no change under any treatment conditions (B). BMDM, bone marrow–derived macrophage; JNK, c-Jun N-terminal kinase; LPS, lipopolysaccharide.
from Cell Signaling Technology for NF-kappaB Pathway Antibody Sampler Kit II