Study On The Effect Of T-614 On LPS-induced NF-κB Signal Transduction In Rat Alveolar Macrophage Cell Line And On Oleic Acid-induced Acute Lung Injury In Rats

Iguratimod (T-614), is a non-steroidal anti-inflammatory drug that hasn't come into the market. Studies show that T-614 has obvious curative effect on rheumatoid arthritis and osteoarthritis, but there are no reports about its effect on lung inflammation. In the present study, we investigate the effect of T-614 on lipopolysaccharide (LPS)-mediated inflammatory process on the level of NF-kB pathway, p38 MAPK pathway and inflammatory factors.In addition, we study the intervene effect of T-614 on animal acute lung injury (ALI) model. 1. The effect of T-614 on LPS-induced rat pulmonary alveolar macrophage cell line NR8383:NR8383 cellls were pretreated with T-614, then were stimulated with LPS. The production of TNF-α in the supernatant of NR8383 was determined with enzyme-linked immunosorbent assay (ELISA).The TNF-α mRNA level was determined by a semi-quantitative PCR assay. NF-κB activity in nuclear extract ofNR8383 was detected with ELISA. The phosphorylation of p38 MAPK was measured by western blotting. The findings indicated that T-614 could inhibit LPS-stimulated mRNA expression and production of TNF-α, and also inhibited the activation of NF-κB, but had no obvious inhibitory effect on the phosphorylation of p38 MAPK. 2. The effect of T-614 on oleic acid-induced ALI rat model:We use an oleic acid-induced ALI rat model to investigate the effect of T-614 in vivo for the first time. T-614 was given before injection of oleic acid at doses of 20, 40, and 80 mg/kg. The changes of lung volume and dynamic lung compliance were real time recorded by anrise2005 pulmonary function test apparatus after injection of oleic acid. We also collected bronchoalveolar lavage fluid(BALF), and measured the protein concentration in BALF. The findings showed that pretreatment of T-614 significantly alleviated oleic acid-induced lung injury accompanied by improvement of lung function and decline of protein concentration in BALF, which suggested that T-614 had a beneficial effect on ALI induced by oleic acid in rat.