[Serum of obesity induce the activation of TLR4/NF-κB signaling pathway on THP-1 cell line].

OBJECTIVE

To investigate the activation of TLR4/NF-κB signaling pathway and the level of inflammation in volunteers with varying degrees of metabolic disorders and the effect of intervention by TLR4 monoclonal antibody.

METHODS

THP-1 cell line was cultured with 10% serum of volunteers with various degrees of metabolic disorders (each 10 cases) for 24, 48 h. TLR4 and NF-κB p65 phosphorylation protein were measured by Western blot. TLR4 mRNA was measured by RT-PCR. The expression of IL-1β and TNF-α were detected by ELISA.

RESULTS

The TLR4 mRNA and protein, the level of NF-κB p65 phosphorylation protein in THP-1 cell line and IL-1β and TNF-α expression in culture supernatant in normal, simple obesity, obesity with hyperglycemia, obesity with hyperlipidemia, obesity with three metabolic disorders groups had statistical significance (P < 0.05). The TLR4 mRNA and protein, the level of NF-κB p65 phosphorylation protein and the IL-1β and TNF-α expression in obesity with three metabolic disorders group were higher than those in other group, and those were time-dependent (P < 0.05). TNF-α expression in normal, simple obesity, obesity with hyperglycemia, obesity with hyperlipidemia, obesity with three metabolic disorders group at 48 h were (222 ± 32), (246 ± 52), (322 ± 32), (322 ± 34) and(490 ± 83)ng/L, respectively, IL-1β (ng/L) were (94 ± 19), (133 ± 19), (174 ± 22), (180 ± 30), (279 ± 38) (P < 0.05).

CONCLUSIONS

The activation of TLR4/NF-κB signaling pathway on THP-1 cell line which is cultured by serum with various degrees of metabolic disorders is different, with the increasing of severity of metabolic disorders, TLR4/NF-κB signaling pathway activation levels also incr.