白藜芦醇对脂多糖诱导的肺泡巨噬细胞微小RNA-146a表达的影响  

Effect of resveratrol on expression of lipopolysaccharide-induced microRNA-146a in alveolarmacrophages

作　　者：曾振国[1] 邵强[1] 李勇[2] 丁成志[1] 卿城[1] 刘芬[1] 詹以安[1] 聂成[1] 揭克敏[4] 龚洪翰[3] 钱克俭[1] 

机构地区：[1]南昌大学第一附属医院重症医学科,南昌330006 [2]南昌大学第一附属医院重症肿瘤科,南昌330006 [3]南昌大学第一附属医院重症影像科,南昌330006 [4]南昌大学医学院生物化学与分子生物学教研室

出　　处：《中华急诊医学杂志》2014年第1期30-33,共4页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金（81060152）;江西省自然科学基金（2009GZY0215）

摘　　要：目的观察白藜芦醇对脂多糖（LPS）诱导肺泡巨噬细胞微小RNA-146a（miR-146a）表达的影响。方法将体外培养的大鼠肺泡巨噬细胞（NR8383）按2×106个／孔接种于六孔板，细胞贴壁90min后，分为磷酸盐缓冲液（PBS）对照组、终质量浓度LPS（1wg／mL）处理组、自藜芦醇（1ug／mL）预处理30min＋终质量浓度LPS（1wg／mL）处理组、白藜芦醇（10wg／mL）预处理30min＋终质量浓度LPS（1gg／mL）处理组。细胞培养6h后，收集细胞和细胞培养上清液，采用实时荧光定量PCR检测细胞中miR-146a的表达变化，采用酶联免疫吸附实验（ELISA）检测细胞培养上清液中TNF-a蛋白的表达变化。结果与PBS对照组相比，LPS处理组细胞中miR．146a和TNF-a蛋白表达量均明显升高[miR-146a表达倍数（5．92±1．57）倍vs．（1．03±0．58）倍，P〈0．01；TNF-a质量浓度（644．2±36．82）pg／mLvs．（26．15．4-13．43）pg／mL，P〈0．01]。与LPS处理组相比，白藜芦醇预处理组（1wg／mL和10ug／mL）细胞中miR-146a表达量均明显升高[miR-146a表达倍数（8．67±1．18）倍、（11．174-1．40）倍VS．（5．92±1．57）倍，P〈0．01]，TNF-a蛋白质量浓度均显著降低[TNF-d质量浓度（447．7±41．48）pg／mL、（345．0±42．54）pg／mLvs．（644．2．4-36．82）pg／mL，P〈0．01]，且相比于较低质量浓度白藜芦醇组（1ug/mL组），较高质量浓度白藜芦醇组（10ug/mL）更能诱导miR-146a的表达、抑制TNF—a蛋白的分泌（P〈0．01）。结论白藜芦醇可以上调肺泡巨噬细胞内miR-146a的表达水平，推测miR-146a参与了白藜芦醇的抗炎过程。Objective To observe the effect of resveratrol on expression of lipopolysaccharide （LPS） -induced microRNA-146a （miR-146a） in alveolar maerophages. Methods Rat alveolar maerophages （ NR8383 ） were seeded at 2 106cell/pore in 6 well plates in vitro and incubated for 90 rain. NR8383 were randomly divided into four groups： control group, NR8383 stimulated with phosphate buffer solution （PBS） ; LPS-stimulated group, NR8383 stimulated with 1 ug/mL of LPS; resveratrol-treated group, NR8383 treated with different concentrations of resveratrol （ 1 txg/mL or 10 ug/mL） for 30 rain, then stimulated with 1 tLg/ mL of LPS. After incubation for 6 h, the cells were harvested and the supematant of cell culture medium was collected. The expression of miR-146a of cells was detected by using fluoroilluminance real-time quantitativePCR device, and the levels of TNF-a protein in the supernatant were assayed by using enzyme-linked immunosorbent assay （ELISA）. Results Compared with control group, the expression of miR-146a and the level of TNF-a were significantly increased in LPS stimulated group （miR-146a＇ s expression folds： 5.92 ±1.57 vs 1.03±0.58, P〈0.01; TNF-ct： 644.2 ±36.82 pg/mL vs. 26. 15 ~ 13.43 pg/mL, P〈 0. 01 ）. Compared with LPS-stimulated group, the expressions of miR-146a were significantly increased in resveratrol-treated groups （1 Ixg/mL and 10 txg/mL） （miR-146a s expression folds： 8.67 ± 1.18, 1l. 17 ±1.40 vs. 5.92 ± 1.57, both P 〈 0.01 ）, but the level of TNF-a in the supernatant was significantly reduced （TNF-a： 447. 7± 41.48 pg/mL, 345.0 ~ 42. 54 pg/mL vs. 644. 2 ± 36. 82 pg/mL, both P 〈 0.01） . Compared with the low concentration of group （1 txg/mL）, the expression of miR-146a were significantly increased and the production of TNF-a in the supernatant of cells was significantly reduced in high concentration of resveratrol group （10 ug/mL） （P 〈0.01）. Conclusions Resveratrol could up- regulate the expression of miR-146a in alveolar ma

关 键 词：白藜芦醇 微小RNA-146a 肺泡巨噬细胞 肿瘤坏死因子-A 

分 类 号：R285.5[医药卫生—中药学]