The Defensive Action of LYRM03 on LPS-Induced Acute Lung Injury by NF-κB/TLR4/NLRP3 Signals

Abstract

The aim of the current investigation was to study the role of 3-amino-2-hydroxy-4-phenyl-valyl-isoleucine (LYRM03) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) and investigate its potential pathogenesis. An LPS-induced ALI model was produced with LPS (5 mg/kg) followed by 24 h of injury. Rats were randomly assigned to 6 groups for <i>in vivo</i> experiments: (1) Sham, (2) LYRM03 (20 mg/kg), (3) LPS, (4) LPS plus LYRM03 (5 mg/kg), (5) LPS plus LYRM03 (10 mg/kg), and (6) LPS plus LYRM03 (20 mg/kg). The rat alveolar macrophage cell line (NR8383) cells were divided into 6 groups for <i>in vitro</i> experiments: (1) Sham, (2) LYRM03 (200 μmol/L), (3) LPS (100 ng/mL), (4) LPS plus LYRM03 (50 μmol/L), (5) LPS plus LYRM03 (100 μmol/L), and (6) LPS plus LYRM03 (200 μmol/L). Further study about siRNA targeting NF-κB p65, TLR4, and NLRP3 to explore the potential mechanism of LYRM03 in the LPS-induced ALI models have been done. Therefore, LYRM03 decreased LPS-induced ALI and NR8383 activation as demonstrated through hematoxylin-eosin staining and western blot analysis <i>in vivo</i> and <i>in vitro</i>. LYRM03 ameliorated the content of protein in bronchoalveolar lavage fluid, myeloperoxidase in the lung and malondialdehyde (MDA) in serum. In addition, LYRM03 ameliorated the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18) in the serum of rats and the supernatant of NR8383 cells. Moreover, LYRM03 significantly inhibited the activities of nuclear factor kappa B (NF-κB), myeloid differentiation factor 88 (MyD88), and toll-like receptor 4 (TLR4). LYRM03 also reduced the increase in the inflammasome, including apoptosis-related speck-like protein containing CARD (ASC), and NOD-like receptor 3 (NLRP3), in LPS-stimulated rats and NR8383 cells. The extent of injury and lung injury scores in the LYRM03 (20 mg/kg) + siRNA targeting NF-κB p65, TLR4, or NLRP3 + LPS-treated rats were higher than that in the LYRM03 (20 mg/kg) + LPS-treated rats. In summary, LYRM03 conferred an intensely lung defensive action on LPS-induced ALI <i>in vivo</i> and <i>in vitro</i>, which could be associated with the abatement of TLR4-induced NLRP3/NF-κB.