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[Differential expression profile of long non-coding RNA in the lipopolysaecharide-induced inflammation of
Zhen Deng1, Fangyi Yao, Jianqing Ye
1Department of Clinical Laboratory, the First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, China (Deng Z, Ye JQ, Xu JQ, Luo Q, Huang ZK); Department of Critical Care Medicine, the First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, China (Qing C); Department of Clinical Laboratory, Jiangxi Province Blood Center, Nanchang 330006, Jiangxi, China (Yao FY). Corresponding author: Huang Zikun,
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/L: 562.93±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 (all P < 0.01). These results indicated that the inflammatory model of human macrophages was constructed successfully. (2) Compared with blank control group, and 1 479 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 (all P < 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.
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