The Role Of Pellino-1 Overexpression On Ubiquitin Of Tumor Necrosis Factor Receptor-Associated Factors 3 And MAPK Signaling Pathway In Endotoxin-Tolerant Kupffer Cells

Objective To investigate the effect of Pellino-1 gene overexpression by lentivirus vector on the ubiquitin of tumor necrosis factor receptor-associated factors 3(TRAF3)and the activation of mitogen-activated protein kinases(MAPK)signaling pathway in endotoxin-tolerant kupffer cells(KCs).Methods Isolated KCs of C57BL/6 mouse were randomly divided into two groups: control group,which transfected with control lentivirus vector for 48 h,then pretreated with 10ng/ml lipopolysaccharide(LPS)for24h,and next treated with 300ng/ml LPS;overexpression group,which transfected with Pellino-1 gene overexpression lentivirus vector for 48 h,then pretreated with 10ng/ml lipopolysaccharide(LPS)for 24 h,and next treated with 300ng/ml LPS.The protein expressions of Pellino-1,K48-Ub,TRAF3,p38,p-p38,c-Jun N-terminal kinase(JNK)and p-JNK were analyzed by western blot.The level of interleukin-1?(IL-1?),tumor necrosis factor-?(TNF-?)and interleukin-10(IL-10)in supernatants were measured by enzyme linked immunosorbent assay(ELISA).Results Compared with control group,the protein expressions of Pellino-1,K48-Ub,p-JNK and p-p38 and the levels of IL-1?(P<0.05),TNF-?(P<0.05)in supernatants was increased in overexpression group,while the protein levels of TRAF3 and the levels of IL-10(P<0.05)in supernatants were decreased.Conclusion Overexpression of Pellino-1 can promote TRAF3 K48 ubiquitination degradation,decrease the protein expression of TRAF3,activate the downstream MAPK signaling pathway,and thus impair the formulation of endotoxin tolerance.