inflammatory response to IL-1b in WT, LDHA KO, NAMPT KO, NFKBIZ KO and FX11 treated chondrocytes

Our goal is to study the impacts of IL-1b treatment on chondrocyte physiology. Furthermore, we knocked out several genes that we believe are important for modulating the inflammatory response. We find that IL-1b treatment induces metabolic reprogramming towards aerobic glycoylsis. We found that inhibition of LDHA using inbhitiro FX11 inhibited the inflammatory response of chondrocytes. We hypothesized that this effect was mediated by interaction with NADH, leading to regulation of IkB-z protein. Thus, deletion of LDHA, NAMPT or NFKBIZ was performed and found to inhibit inflammatory response genes induced by IL-1b, with similar gene expression changes amongst the three suggesting that they may work in the same pathway. Chondrocytes were isolated from sterna of WT animals and treated with IL-1b (10 ng/mL) in the presence or absence of FX11 (40 uM) for 24 hours. For other samples, chondrocytes were isolated from sterna of LDHA flox, NAMPT flox or NFKBIZ flox pups. Chondrocytes were adenovirally transduced with GFP or Cre expressing vectors to delete genes. Cells were then treated with IL-1b for 24 hours. No passaging was perfomred on chondrocytes, all cultures were performed at normoxia.