金雀异黄素对脂多糖诱导人肺泡上皮细胞A549炎症及凋亡的影响
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摘要
目的观察金雀异黄素对脂多糖(LPS)诱导人肺泡上皮细胞A549发生炎症与凋亡的影响,探讨其改善肺损伤的作用机制。方法 CCK-8法检测不同浓度金雀异黄素和/或LPS干预细胞12 h后的活力;RT-PCR检测金雀异黄素对LPS诱导的炎性因子白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)的表达;TUNEL法检测细胞凋亡;Western blot检测信号通路的变化。结果金雀异黄素(1、5、10μmol/L)对细胞活力无明显影响,LPS(1μg/m L)可显著降低细胞活力,而金雀异黄素与LPS共同干预则细胞活力呈浓度依赖性升高;RT-PCR结果显示,LPS可显著提高炎性因子的基因表达,而金雀异黄素则可呈浓度依赖和时间依赖抑制其表达;TUNEL染色结果显示,金雀异黄素(10μmol/L)与LPS联合干预12 h可显著减少LPS诱导的细胞凋亡;Western blot结果显示,金雀异黄素(10μmol/L)与LPS(1μg/m L)协同刺激A549细胞30 min可显著抑制LPS诱导的IκBα、p65信号通路的磷酸化水平。结论金雀异黄素可抑制LPS诱导的人肺泡上皮细胞发生炎症和凋亡,从而发挥保护作用。
Objective To investigate the effects of Genistein(GEN) acting on human lung adenocarcinoma A549 cells induced by LPS; To discuss its action of mechanism for improving injuries of lung. Methods The activity of cell treated with different concentrations of Genistein was detected by CCK-8 method and / or LPS intervention for 12 h. The expressions of inflammatory cytokines IL-1β, IL-6, and TNF-α induced by LPS were detected by RT-PCR. TUNEL was used to detect apoptosis, and Western blot was used to detect the changes of signal pathway. Results GEN(1, 5, 10 μmol/L) alone had no effects on cell activity; LPS(1 μg/m L) reduced cell viability significantly, while co-treated the A549 cells with GEN and LPS could reverse this condition in a concentration-dependent manner; RT-PCR results showed that LPS could significantly increase the level of gene expression of inflammatory factors, while GEN could inhibited this phenomenon in both concentration-and time-dependent manner; the results of TUNEL staining showed that GEN(10 μmol/L) combined with LPS for 12 h could significantly decrease the apoptosis rate; the results of Western blot showed that GEN possibly inhibited the inflammation and apoptosis through inhibiting the phosphorylation of IκBα, p65 signaling pathways. Conclusion GEN has anti-inflammation and anti-apoptosis effects on A549 cells induced by LPS, so as to play a protective rate.
Objective To investigate the effects of Genistein(GEN) acting on human lung adenocarcinoma A549 cells induced by LPS; To discuss its action of mechanism for improving injuries of lung. Methods The activity of cell treated with different concentrations of Genistein was detected by CCK-8 method and / or LPS intervention for 12 h. The expressions of inflammatory cytokines IL-1β, IL-6, and TNF-α induced by LPS were detected by RT-PCR. TUNEL was used to detect apoptosis, and Western blot was used to detect the changes of signal pathway. Results GEN(1, 5, 10 μmol/L) alone had no effects on cell activity; LPS(1 μg/m L) reduced cell viability significantly, while co-treated the A549 cells with GEN and LPS could reverse this condition in a concentration-dependent manner; RT-PCR results showed that LPS could significantly increase the level of gene expression of inflammatory factors, while GEN could inhibited this phenomenon in both concentration-and time-dependent manner; the results of TUNEL staining showed that GEN(10 μmol/L) combined with LPS for 12 h could significantly decrease the apoptosis rate; the results of Western blot showed that GEN possibly inhibited the inflammation and apoptosis through inhibiting the phosphorylation of IκBα, p65 signaling pathways. Conclusion GEN has anti-inflammation and anti-apoptosis effects on A549 cells induced by LPS, so as to play a protective rate.
引文
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[9]马莹莹莹,张育,许金鑫鑫,等.金雀异黄素素对CIA大鼠促炎炎和抗炎因子分泌泌的影响响[J].实用药物与与临床,2014,17(10):1247-1251..
[10]冯冯赞杰,卢志顺,刘刘洋,等.金雀异黄黄素抑制人血管平平滑肌细胞中核因因子κBB的激活[J].中国国动脉硬化杂志,22009,17(5):3633-366.
[2]XIIE X,SUN S,ZHONG W,e t al.Zingeroo ne attenuates s lipopp olysaccharide--induced acute lung injury i n mice[J].Int t Immunn opharmacol,20114,19(1):103-109.
[3]LOUUT,JIANG W,XUU D,et al.Inhi ibitory effects of polydatin on n lipop olysaccharide-s timulated RAW264.7 cells[J]..Inflammation,2015,,38(3):1213-12220.
[4]TAOOW,SU Q,WANGH,et al.Platy ycodin D attenu ates acute lung g injurr y by suppressi ng apoptosis an nd inflammationn in vivo and in n vitroo[J].Int Immun nopharmacol,2015,27(1):138-1447.
[5]BAO S,ZOU Y,WANG B,et a l.Ginsenosidee Rg1 improves s lipopp olysaccharide--induced acute lung injury by inhibiting g inflaa mmatory respo onses and mod ulating infiltt ration of M22macroo phages[J].Int t Immunopharmac ol,2015,28(1):429-434.
[6]LIY,WU Q,DENG Y,et al.D(-)-Sa licin inhibits the LPS-induced d inflaa mmation in RRAW264.7 cells and mouse m odels[J].Int t Immunn opharmacol,20115,26(2):286-294.
[7]王玲玲,刘辉国.金雀异异黄素对肺泡Ⅱ型型上皮细胞发生上上皮-间质转化的的影响[J].中国中医药信信息杂志,2016,23(10):63-66.
[8]金永永生,刘超美.金雀雀异黄素的药理作作用[J].国外医药药:植物药分册,2002,,17(5):190-1933.
[9]马莹莹莹,张育,许金鑫鑫,等.金雀异黄素素对CIA大鼠促炎炎和抗炎因子分泌泌的影响响[J].实用药物与与临床,2014,17(10):1247-1251..
[10]冯冯赞杰,卢志顺,刘刘洋,等.金雀异黄黄素抑制人血管平平滑肌细胞中核因因子κBB的激活[J].中国国动脉硬化杂志,22009,17(5):3633-366.