微小RNA-132在脂多糖诱导大鼠肺泡巨噬细胞炎症反应中的表达变化

The expression changes in microRNA-132 in the lipopolysaccharide-induced inflammation of rat alveolarmacrophages

目的观察脂多糖（LPS）诱导大鼠肺泡巨噬细胞炎症反应后微小RNA-132（miR-132）的动态变化，以初步探讨miR-132在肺泡巨噬细胞炎症反应中的作用。方法将体外去致热源培养的大鼠肺泡巨噬细胞株NR8383分为空白对照组和以终浓度1mg／LLPS刺激3、6、12及24h组，分别收集各时间点上清液及细胞，采用酶联免疫吸附试验（ELISA）检测上清液中炎症因子肿瘤坏死因子-0I（TNF-仅）、白细胞介素（IL-113和IL-6）的含量，采用实时定量聚合酶链反应（PCR）检测细胞中miR-132的表达。结果与空白对照组相比，LPS刺激大鼠肺泡巨噬细胞后3h上清液中TNF-α．含量（ng／L：364．83±46．29比34．07±8．62，P〈O．01）、IL-113含量（ng／L：153．83±43．67比32．334-10．62，P〈0．05）、IL-6含量（ng／L：183．854-43．52比42．62±11．21，P〈0．05）即明显升高，随时间延长均逐渐升高至12h达峰值（TNF-α：605．09±57．13，IL-1B：377．09±28．55，IL-6：558．04±77．45，均P〈O．01），24h时均有所下降（TNF-α：281．95±41．61，IL-18：263．17±51．36，IL-6：438．74±79．94），但仍显著高于空白对照组（均P〈0．01）。LPS刺激后3h，miR-132表达水平较空白对照组上调了（1．12±0．11）倍（P=0．995），6、12、24hmiR-132表达较空白对照组分别上调了（5．98±0．65）、（7．64±0．53）和（8．92±0．83）倍（均P〈0．01）。结论LPS诱导大鼠肺泡巨噬细胞炎症反应后，miR-132表达随时间延长逐步上调，其可能参与调控大鼠肺泡巨噬细胞炎症反应。

Objective 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. Methods 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-1 [3 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. Results 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-113 （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〈O.05） were all increased significantly at 3 hours post LPS stimulation compared with blank control group, reached the peak at 12 hours （TNF-a： 605.09 ± 57.13, IL-113：377.09 ± 28.55, IL-6：558.04 -＋ 77.45, all P〈0.01 ）, and descended at 24 hours （TNF-α： 281.95 ± 41.61, IL-113：263.17 ± 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=0.9953, 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]. Conclusion The expression of miR-132 increased gradually over time after LPS-induced inflammation of rat alveolar maerophages, suggesting that miR-132 may be involved in regulation of rat alveolar macrophages inflammatory response.