Objective To analyze the expression profile of
long non-coding RNA (
lncRNA) in the lipopolysaecharide (LPS)-induced
inflammation of
monocyte-derived macrophages.
Methods Peripheral blood mononuclear cells were derived from healthy
donor and induced into
macrophages. The
macrophages were divided into blank
control group and LPS (1 mg/L) stimulated 12 hours group.
Culture supernatants and
cell pellets were harvested in each group,
enzyme linked immunosorbent assay (
ELISA) was used to assay the
production changes of
interleukins (
IL-1β and
IL-6), and
tumor necrosis factor-α (TNF-α) in the supernatant. The
technique of
lncRNA microarray was used to test the
lncRNA expression profile in LPS-induced
inflammation of
macrophages and control
macrophages. The raw data of
lncRNA were pretreated for normalization. Five
lncRNA expressions were validated by real-
time quantitative
reverse transcription-
polymerase chain reaction (qRT-
PCR). Furthermore, qRT-
PCR was used to detect the expression of NR_028034 in
macrophages after LPS-induced
inflammation.Results ① The contents of
IL-1β (ng/L562.93±61.17 vs. 59.74±15.68),
IL-6 (ng/L 702.46±92.31 vs. 71.66±18.25) and TNF-α (ng/L 794.50±63.89 vs. 85.12±22.07) in the LPS group were significantly higher than those in the blank
control group (allP < 0.01). These results indicated that the inflammatory model of
human macrophages was constructed successfully. ② Compared with blank
control group, and 1479
lncRNA which have more than 2 folds variation and significant difference (P < 0.05) by
statistical analysis was defined as
lncRNA with differential expression. Among these
lncRNA, LPS group showed 953 up- regulated and 526 down- regulated
genes by 2 folds and 49 up- regulated and 35 down- regulated
genes by 5 folds. ③ qRT-
PCR results were generally consistent with the microarray data. ④ The expression of NR_028034 was increased by (4.41±0.65), (11.56±2.04), (18.58±1.36) folds compared with blank
control group at 3, 6, 12 hours after LPS stimulation (allP < 0.01).Conclusions These data show a significantly altered
lncRNA expression profile in the LPS-induced
inflammation of
monocyte-derived macrophages, suggesting that
lncRNA may be involved in
regulation of
macrophages inflammatory response.