摘要
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.
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2014年第2期80-83,共4页
Chinese Critical Care Medicine
基金
国家自然科学基金(81101410)
国家自然科学基金(81160233)
江西省自然科学基金(20122BAB205002)
