Role of heme oxygenase-1 on lipopolysaccharide-induced activation of NLRP3 inflammasomes in alveolar macrophages of rats / 中华麻醉学杂志

ABSTRACT

Objective To evaluate the role of heme oxygenase-1 (HO-1) on lipopolysaccharide (LPS)-induced activation of NOD-like receptor pyrin domain-containing 3 (NLRP3) inflammasomes in alveolar macrophages of rats.Methods NR8383 cells of rat alveolar macrophages cultured in vitro were seeded in 96-well plates at a density of 4× 104 cells/ml and divided into 4 groups (n =48 each) using a random number table method:control group (group C),group LPS (group L),LPS+HO-1 siRNA group (group L+HO-1 siRNA),and LPS+Con siRNA group (group L+Con siRNA).The cells were cultured in normal culture atmosphere in group C.NR8383 cells were stimulated with 10 μg/ml LPS in L,L+HO-1 siRNA and L+Con siRNA groups.Cells were transfected with HO-1 siRNA at 48 h before stimulation with LPS in group L+HO-1 siRNA and with Con siRNA at 48 h before stimulation with LPS in group L+Con siRNA.The ceils were collected and incubated for 24 h after stimulation with LPS for measurement of the cell viability,expression of HO-1 and NLRP3 mRNA (by real-time polymerase chain reaction),expression of HO-1,NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC)and caspase-1 (by Western blot),and concentrations of interleukin-1beta (IL-1β) and interleukin-18 (IL-18) in the supernatant (by enzyme-linked immunosorbent assay).Results Compared with group C,the cell viability was significantly decreased,the expression of HO-1 and NLRP3 protein and mRNA,ASC and caspase-1 was up-regulated,and the IL-1β and IL-18 concentrations were increased in L,L+HO-1 siRNA and L+Con siRNA groups (P＜0.05).Compared with group L,the cell viability was significantly decreased,the expression of HO-1 protein and mRNA was down-regulated,and the expression of NLRP3 protein and mRNA,ASC and caspase-1 was up-regulated,and the IL-1β and IL-18 concentrations were increased in group L+HO-1 siRNA (P＜0.05),and no significant change was found in the parameters mentioned above in group L+Con siRNA (P＞0.05).Conclusion HO-1 is involved in LPS-induced activation of NLRP3 inflammasomes in alveolar macrophages of rats.