The mechanism underlying
lipopolysaccharide (LPS)-induced aberrant proliferation of lung fibroblasts in Gram-negative bacilli-associated
pulmonary fibrosis is unknown. High-mobility group box 1 (
HMGB1) is a ubiquitous
nuclear protein that is released from the nuclei of lung fibroblasts after LPS stimulation. It can exasperate LPS-induced
inflammation and hasten cell proliferation. Thus, this study investigated the effects of LPS- and/or HMGB1-stimulating murine lung fibroblasts on gene expression using various assays in vitro. Thiazolyl-
diphenyl-tetrazolium
bromide (MTT) assay data showed that either LPS or
HMGB1 could induce lung fibroblast proliferation. Endogenous
HMGB1 secreted from lung fibroblasts was detected by
enzyme-linked
immunosorbent assay (ELISA) 48 h after LPS stimulation. Pretreatment with an anti-HMGB1 antibody inhibited the proliferative effects of LPS on lung fibroblasts.
DNA microarray data showed that the NF-κB signaling genes were upregulated in cells after stimulated with LPS,
HMGB1, or both. Secretion of
matrix metalloproteinase (MMP)-2 and MMP-9, and
tissue inhibitor of metalloproteinase 2 (TIMP-2) was significantly upregulated
after treatment with LPS,
HMGB1, or their combination. However, an NF-κB inhibitor was able to downregulate levels of these
proteins. In addition, levels of
Toll-like receptor 4 (TLR4),
Toll-like receptor 2 (TLR2), and receptors for
advanced glycation end products (RAGE)
mRNA and
proteins were also upregulated in these cells after LPS treatment and further upregulated by LPS plus
HMGB1. In conclusion, the data from the current study demonstrate that LPS-induced lung fibroblast secretion of endogenous
HMGB1 can augment the proproliferative effects of LPS and, therefore, may play a key role in exacerbation of
pulmonary fibrosis. The underlying molecular mechanisms are related to the activation of the TLR4/NF-κB signaling pathway and its downstream targets.