Fen Liu, Rong Jiang, Zhenguo Zeng, Ning Zhao, Liang Xia, Cheng Nie, Kejian Qian
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2014 Feb;26(2):80-3.
To observe the kinetic changes in microRNA-132 (miR-132) expression in rat alveolar macrophages after lipopolysaccharide (LPS)-induced inflammation, and to investigate initially on the role of miR-132 in alveolar macrophages inflammatory response.
The rat alveolar macrophages NR8383 cultured without pyrogen in vitro were divided into blank control group and LPS (1 mg/L) stimulated 3, 6, 12, 24 hours groups. Culture supernatants and cell pellets were collected at each time point respectively. Enzyme-linked immunosorbent assay (ELISA) was used to assay the production changes in tumor necrosis factor-α (TNF-α), interleukins (IL-1β and IL-6) in the supernatant. Real-time quantitative polymerase chain reaction (PCR) was used to detect the expression of miR-132 in the cells.
After stimulating rat alveolar macrophages with LPS, the production of TNF-α (ng/L: 364.83 ± 46.29 vs. 34.07 ± 8.62, P<0.01), IL-1β (ng/L: 153.83 ± 43.67 vs. 32.33 ± 10.62, P<0.05) and IL-6 (ng/L: 183.85 ± 43.52 vs. 42.62 ± 11.21, P<0.05) were all increased significantly at 3 hours post LPS stimulation compared with blank control group, reached the peak at 12 hours (TNF-α: 605.09 ± 57.13, IL-1β: 377.09 ± 28.55, IL-6: 558.04 ± 77.45, all P<0.01), and descended at 24 hours (TNF-α: 281.95 ± 41.61, IL-1β: 263.17 ± 51.36, IL-6: 438.74 ± 79.94) but the levels remained significantly higher than blank control group (all P<0.01). The expression of miR-132 started to rise at 3 hours after LPS stimulation compared with blank control group [(1.12 ± 0.11) folds, P=0.995], and presented a gradual increasing trend at 6, 12, 24 hours [(5.98 ± 0.65), (7.64 ± 0.53), (8.92 ± 0.83) folds, all P<0.01].
The expression of miR-132 increased gradually over time after LPS-induced inflammation of rat alveolar macrophages, suggesting that miR-132 may be involved in regulation of rat alveolar macrophages inflammatory response.
观察脂多糖（LPS）诱导炎症后大鼠肺泡巨噬细胞中微小RNA-132（miR-132）表达的动态变化，并初步探讨miR-132在肺泡巨噬细胞炎症反应中的作用。
将体外无热源培养的大鼠肺泡巨噬细胞NR8383分为空白对照组和LPS（1 mg/L）刺激3、6、12、24小时组。分别在各时间点收集细胞培养上清液和细胞沉淀。采用酶联免疫吸附测定（ELISA）法检测上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素（IL-1β和IL-6）的产生变化。采用实时定量聚合酶链反应（PCR）法检测细胞中miR-132的表达。
LPS刺激大鼠肺泡巨噬细胞后，与空白对照组相比，LPS刺激后3小时TNF-α（ng/L：364.83±46.29比34.07±8.62，P<0.01）、IL-1β（ng/L：153.83±43.67比32.33±10.62，P<0.05）和IL-6（ng/L：183.85±43.52比42.62±11.21，P<0.05）的产生均显著增加，在12小时达到峰值（TNF-α：605.09±57.13，IL-1β：377.09±28.55，IL-6：558.04±77.45，均P<0.01），并在24小时下降（TNF-α：281.95±41.61，IL-1β：263.１7±51.36，IL-6：438.74±79.94），但水平仍显著高于空白对照组（均P<0.01）。与空白对照组相比，LPS刺激后3小时miR-132的表达开始升高[（1.12±0.11）倍，P=0.995]，并在6、12、24小时呈逐渐升高趋势[（5.98±0.65）、（7.64±0.53）、（8.92±0.83）倍，均P<0.01]。
LPS诱导大鼠肺泡巨噬细胞炎症后，miR-132的表达随时间逐渐增加，提示miR-132可能参与大鼠肺泡巨噬细胞炎症反应的调节。
