苹果多糖抑制脂多糖引起宫颈癌Hela细胞的TLR-4/NF-κB通路活化
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摘要
目的:探讨苹果多糖(AP)对脂多糖(LPS)活化人宫颈癌细胞Hela中TLR-4/NF-κB通路的影响及相关机制。方法:培养Hela细胞,将其分为对照组、LPS处理组(1μg/ml)、AP(0.5 mg/ml)+LPS处理组(1μg/ml),采用免疫荧光法观察NF-κB在细胞质、细胞核内的分布情况,免疫印迹法检测Hela细胞中IκB-α,磷酸化IκB-α,细胞核中NF-κB以及细胞膜上TLR-4的蛋白变化。结果:AP可抑制LPS刺激引起的Hela细胞的NF-κB核转位,可有效抑制IκB-α降解及IκB-α的磷酸化,可降低细胞膜表面的TLR-4表达。结论:AP可能通过抑制脂多糖活化TLR-4分子来抑制NF-κB通路的激活。
Objective: To assess the function of apple polysaccharides( AP) on TLR-4/NF-κB pathway and the possible mechanisms in lipopolysaccharide( LPS) treated human cervical cancer cells Hela. Methods: Hela cells were divided randomly into control group,LPS-treated groups( 1 μg/ml),AP( 0. 5 mg/ml) plus LPS-treated group( 1 μg/ml). Then,the distribution of NF-κB was assessed by immunofluorescence. The expression of IκB-α,phosphorylated IκB-α,NF-κB as well as TLR-4 in the membrane,were measured using Western blot. Results:AP could inhibit LPS-stimulated nuclear translocation of NF-κB,degradation of IκB-α,phosphorylation of IκB-α,and reduce TLR-4 expression. Conclusion: AP may suppress the activation of NF-κB signaling pathway through inhibition of LPS-stimulated TLR-4 expression.
Objective: To assess the function of apple polysaccharides( AP) on TLR-4/NF-κB pathway and the possible mechanisms in lipopolysaccharide( LPS) treated human cervical cancer cells Hela. Methods: Hela cells were divided randomly into control group,LPS-treated groups( 1 μg/ml),AP( 0. 5 mg/ml) plus LPS-treated group( 1 μg/ml). Then,the distribution of NF-κB was assessed by immunofluorescence. The expression of IκB-α,phosphorylated IκB-α,NF-κB as well as TLR-4 in the membrane,were measured using Western blot. Results:AP could inhibit LPS-stimulated nuclear translocation of NF-κB,degradation of IκB-α,phosphorylation of IκB-α,and reduce TLR-4 expression. Conclusion: AP may suppress the activation of NF-κB signaling pathway through inhibition of LPS-stimulated TLR-4 expression.
引文
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[3]Ma YZ,Zhang YJ.Research progress of Toll like receptor 4 and 9in cervical cancer[J].Shandong Medicine,2013,53(41):97-99.[马玉着,章一涓.Toll样受体4和9与宫颈癌的相关研究进展[J].山东医药,2013,53(41):97-99.]
[4]Wang Y,Weng Y,Shi Y,et al.Expression and functional analysis of Toll-like receptor 4 in human cervical carcinoma[J].J Membr Biol,2014,247(7):591-599.
[5]Yan W,Xu FG,Liu AW,et al.Effect of Toll like-receptor 4 on proliferation of cervical cancer cells by NF-κB signaling pathway[J].Chin J Pathophysiol,2015,31(2):301-307.[颜伟,许芳根,刘安文,等.TLR-4通过NF-κB信号途径对宫颈癌细胞增殖的影响[J].中国病理生理杂志,2015,31(2):301-307.]
[6]Li Y,Fan L,Sun Y,et al.An apple oligogalactan suppresses endotoxin-induced cyclooxygenase-2 expression by inhibition of LPS pathways[J].Int J Biol Macromol,2013,61:75-81.
[7]Liu LN,Guo ZW,Zhang Y,et al.Polysaccharide extracted from Rheum tanguticum prevents irradiation-induced immune damage in mice[J].Asian Pac J Cancer Prev,2012,13(4):1401-1405.
[8]Tang Y,Sun Y,Li YH,et al.Effects of apple polysacchsrides on the apoptsis of human colon cancer SW-1116 cells and the possible mechanisms[J].Mil Med J S Chin,2012,26(3):229-232.[唐源,孙阳,李宇华,等.苹果多糖对SW-1116结肠癌细胞凋亡的影响及机制研究[J].华南国防医学杂志,2012,26(3):229-232.]
[9]Janeway CA Jr.Approaching the asymptote evolution and revolution inimmunology[J].Cold Spring Harb Syrup Quant Biol,1989,54(1):1-13.
[10]Lemaitre B,Nicolas E,Michaut L,et al.The dorsoventral regulatory gene cassette spatzle/Toll/cactus controls the potent antifungal response in Drosophila adults[J].Cell,1996,86(6):973-983.
[11]Medzhitov R,Preston-Hurlbua P,Janeway CA Jr.A human homologne of the Drosophila toll protein signals activation of adaptive immunity[J].Nature,1997,388(6640):394-397.
[12]Roy A,Srivastava M,Saqib U,et al.Potential therapeutic targets for inflammation in toll-like receptor 4(TLR-4)-mediated signaling pathways[J].Int Immunopharmacol,2016,40:79-89.
[13]Marey MA,Yousef MS,Kowsar R,et al.Local immune system in oviduct physiology and pathophysiology:attack or tolerance[J]?Domest Anim Endocrinol,2016,56(Suppl):S204-S211.
[14]Mitchell S,Vargas J,Hoffmann A.Signaling via the NF-κB system[J].Wiley Interdiscip Rev Syst Biol Med,2016,8(3):227-241.
[15]Rakoff-Nahoum S,Medzhitov R.Regulation of spontaneous intestinal tumorigenesis through the adaptor protein My D88[J].Science,2007,317(5834):124-127.