小白菊内酯通过NF-κB信号通路对LPS诱导的气道上皮细胞IL-8分泌水平影响的研究
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摘要
目的探究小白菊内酯(PTL)对脂多糖(LPS)诱导的气道上皮细胞IL-8的分泌水平影响以及具体的作用机制。方法 CCK8法检测0μg/ml、5μg/ml、10μg/ml、20μg/ml、40μg/ml、80μg/ml的LPS对气道上皮细胞的毒性作用,RT-q PCR和Western blot检测不同浓度LPS对IL-8表达水平的影响。添加0μmol/L、10μmol/L、20μmol/L、40μmol/L、80μmol/L、160μmol/L PTL后,CCK8法检测细胞活性变化; RT-q PCR和Western blot检测白介素8(IL-8)表达水平的变化; Western blot检测PTL对NF-κB p65和IκBα蛋白表达水平的影响。结果 LPS呈时间和浓度依赖性抑制气道上皮细胞活性; LPS呈时间依赖性促进IL-8表达水平;添加LPS能够使IκB表达下调,NF-κB p65表达上调。添加适当浓度的PTL能够缓解LPS对细胞的毒性影响,使IL-8表达下调,具有浓度依赖性;并且促使IκB下调,NF-κB p65上调。结论小白菊内酯能够通过NF-κB信号通路抑制气道上皮细胞炎症因子IL-8的分泌水平。
Objective To investigate the effect and mechanism of parithenolide on the level of inflammatory factor in LPS-induced airway epithelium cells. Methods CCK8 was performed to detect the cell activity of 16 HBE cells induced by LPS at the concentration of 0 μg/ml、5 μg/ml、10 μg/ml、20 μg/ml、40 μg/ml、and 80 μg/ml,RT-qPCR and Western blot were used to detect the eapression level of IL-8. After 0μmol/L,10 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L and 160 μmol/L PTL was added to LPS-induced cells,the cell activity was tetected by CCK8,the changes of IL-8 expression were detected by RT-qPCR and Western blot,the expression of NF-κB p65 and IκBα protein was detected by Western blot. Results LPS inhibited the activity of airway epithelial cells in time and concentration dependence. LPS promoted the expression of IL-8 in a time-dependent manner,and LPS could reduce the expression level of IκB and increase the NF-κB p65 expression level. PTL at a proper concentration could alleviate the toxic effects of LPS on cells,reduce the expression of IL-8 in time-dependent and concentration-dependent manner,promote down-regulation of IκB and up-regulation of NF-κB p65. Conclusion Pyrethroid could inhibit the secretion of inflammatory factor IL-8 in airway epithelial cells through the NF-κB signaling pathway.
Objective To investigate the effect and mechanism of parithenolide on the level of inflammatory factor in LPS-induced airway epithelium cells. Methods CCK8 was performed to detect the cell activity of 16 HBE cells induced by LPS at the concentration of 0 μg/ml、5 μg/ml、10 μg/ml、20 μg/ml、40 μg/ml、and 80 μg/ml,RT-qPCR and Western blot were used to detect the eapression level of IL-8. After 0μmol/L,10 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L and 160 μmol/L PTL was added to LPS-induced cells,the cell activity was tetected by CCK8,the changes of IL-8 expression were detected by RT-qPCR and Western blot,the expression of NF-κB p65 and IκBα protein was detected by Western blot. Results LPS inhibited the activity of airway epithelial cells in time and concentration dependence. LPS promoted the expression of IL-8 in a time-dependent manner,and LPS could reduce the expression level of IκB and increase the NF-κB p65 expression level. PTL at a proper concentration could alleviate the toxic effects of LPS on cells,reduce the expression of IL-8 in time-dependent and concentration-dependent manner,promote down-regulation of IκB and up-regulation of NF-κB p65. Conclusion Pyrethroid could inhibit the secretion of inflammatory factor IL-8 in airway epithelial cells through the NF-κB signaling pathway.
引文
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[3] Islam MN,Ishita IJ,Jin SE,et al. Anti-inflammatory activity of edible brown alga Saccharina japonica and its constituents pheophorbide a and pheophytin a in LPS-stimulated RAW 264. 7 macrophage cells[J]. Food Chem Toxicol,2013,55(3):541-548.
[4] Qin X,Qiu C,Zhao L. Lysophosphatidylcholine perpetuates macrophage polarization toward classically activated phenotype in inflammation[J]. Cell Immunol,2014,289(1-2):185-190.
[5] Schepetkin IA,Kirpotina LN,Mitchell PT,et al. The natural sesquiterpene lactones arglabin,grosheimin,agracin,parthenolide,and estafiatin inhibit T cell receptor(TCR)activation[J]. Phytochemistry,2018,146:36-46.
[6] Ren Y,Gallucci JC,Li X,et al. Crystal Structures and Human Leukemia Cell Apoptosis Inducible Activities of Parthenolide Analogues Isolated from Piptocoma rufescens[J]. J Nat Prod,2018,81(3):554-561.
[7] Li H,Lu H,Lv M,et al. Parthenolide facilitates apoptosis and reverses drug-resistance of human gastric carcinoma cells by inhibiting the STAT3 signaling pathway[J]. Oncol Lett,2018,15(3):3572-3579.
[8] Zhang M,Liu RT,Zhang P,et al. Parthenolide inhibits the initiation of experimental autoimmune neuritis[J]. J Neuroimmunol,2017,305:154-161.
[9] Onozato T,Nakamura CV,Cortez DA,et al. Tanacetum vulgare:antiherpes virus activity of crude extract and the purified compound parthenolide[J]. Phytother Res,2007,23(6):791-796.
[10] Zhu Z,Gu Y,Zhao Y,et al. GYF-17,a chloride substituted 2-(2-phenethyl)-chromone,suppresses LPS-induced inflammatory mediator production in RAW264. 7 cells by inhibiting STAT1/3 and ERK1/2 signaling pathways[J]. Int Immunopharmacol,2016,35:185-192.
[11] Grammas P,Ovase R. Inflammatory factors are elevated in brain microvessels in Alzheimer's disease[J]. Neurobiol Aging,2001,22(6):837-842.
[12] Aghadavod E,Khodadadi S,Baradaran A,et al. Role of Oxidative Stress and Inflammatory Factors in Diabetic Kidney Disease[J]. Iran J Kidney Dis,2016,10(6):337-343.
[13] Yang Y,Gao SG,Zhang FJ,et al. Effects of osteopontin on the expression of IL-6 and IL-8 inflammatory factors in human knee osteoarthritis chondrocytes[J]. Cell Biochem Biophys,2014,70(1):703.
[14] Wang JE,Jrgensen PF,Ellingsen EA,et al. Peptidoglycan primes for LPS-induced release of proinflammatory cytokines in whole human blood[J]. Shock,2001,16(3):178-182.
[15] Jeyamohan S,Moorthy RK,Kannan MK,et al. Parthenolide induces apoptosis and autophagy through the suppression of PI3K/Akt signaling pathway in cervical cancer[J]. Biotechnol Lett,2016,38(8):1-10.
[16] Zhang S,Lin ZN,Yang CF,et al. Suppressed NF-κB and sustained JNK activation contribute to the sensitization effect of parthenolide to TNF-α-induced apoptosis in human cancer cells[J]. Carcinogenesis,2004,25(11):2191-2199.
[17] Saadane A,Eastman J,Berger M,et al. Parthenolide inhibits ERK and AP-1 which are dysregulated and contribute to excessive IL-8 expression and secretion in cystic fibrosis cells[J]. J Inflamm(Lond),2011,8:26.
[18] Catrysse L,Loo GV. Inflammation and the Metabolic Syndrome:The Tissue-Specific Functions of NF-κB[J]. Trends in Cell Biol,2017,27(6):417-429.
[19] Vageli DP,Doukas SG,Sasaki CT. Inhibition of NF-κB prevents the acidic bile-induced oncogenic mRNA phenotype,in human hypopharyngeal cells[J]. Oncotarget,2018,9(5):5876-5891.
[20] Li X,Zhao Y,Tian B,et al. Modulation of gene expression regulated by the transcription factor NF-κB/Rel A[J]. J Biol Chem,2014,289(17):11927-11944.
[21] Virlos I,Mazzon E,Serraino I,et al. Pyrrolidine dithiocarbamate reduces the severity of cerulein-induced murine acute pancreatitis[J].Shock,2003,20(6):544-550.