柚皮素对RSV A2株感染引起气道黏液高分泌的抑制作用

摘要
图/表
参考文献
相关文章 (2)

全文: PDF (1732 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的：探讨柚皮素（Nar）对呼吸道合胞病毒（RSV）感染引起的气道黏液高分泌的作用。方法：通过RSV A2株感染人肺腺癌上皮细胞构建气道黏液高分泌细胞模型，在RSV感染及Nar干预作用下，采用实时荧光定量PCR、ELISA和细胞免疫荧光技术检测各组细胞黏蛋白（MUC）5AC的表达情况。结果：RSV感染R1组[感染复数（MOI）=0.5]、R2组（MOI=1.0）、R3组（MOI=5.0）和对照组（C组）MUC5AC mRNA相对表达量分别为1.45±0.10、2.67±0.09、4.82±0.27和1，各组MUC5AC蛋白的表达量分别为（38.61±0.41）μg/L、（44.11± 0.96）μg/L、（51.01±0.48）μg/L和（34.38±0.95）μg/L，RSV感染组和C组相比差异均有统计学意义（P＜0.05）。R2组和不同浓度Nar干预组N1组（Nar 30 μmol/L+RSV MOI=1.0）、N2组（Nar 100 μmol/L+RSV MOI=1.0）、N3组（Nar 30 μmol/L）、N4组（Nar 100 μmol/L）及DMSO组（D组）的MUC5AC mRNA水平依次为2.52±0.10、2.31±0.06、1.90±0.01、0.99±0.04、0.85±0.03、1.20±0.03，MUC5AC蛋白水平依次为（46.72±0.87）μg/L、（43.60±0.45）μg/L、（40.55±0.45）μg/L、（38.73±0.55）μg/L、（37.08±1.35）μg/L、（37.12±0.91）μg/L，C组MUC5AC蛋白水平为（37.62±1.18）μg/L，与C组相比，R2组MUC5AC mRNA和蛋白水平显著增加（P＜0.05），N1、N2组与R2组相比，MUC5AC mRNA和蛋白水平显著降低，随Nar浓度增加，抑制效果更明显，呈剂量相关性（P＜0.05），N3、N4组与C组相比，MUC5AC mRNA和蛋白表达水平下降，但差异无统计学意义（P＞0.05），D组与C组差异无统计学意义（P＞0.05）。荧光显微镜观察结果显示RSV感染后细胞质内MUC5AC蛋白表达增加，而Nar可下调其表达。结论：RSV A2株感染A549细胞会导致MUC5AC mRNA和蛋白表达增加，Nar通过抑制其表达，抑制RSV感染引起的气道黏液高分泌。

	服务
	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	孟珊珊1
	2
	吕芳芳1
	胡晓光1
	张海邻1
	李昌崇1

关键词 ：柚皮素, 呼吸道合胞病毒, 气道黏液高分泌, 黏蛋白5AC

Abstract：Objective: To investigate the expression levels of MUC5AC protein and mRNAin mucous hypersecretion model induced by respiratory syncytial virus (RSV), and explore the effect of Naringenin (Nar) on mucous hypersecretion induced by RSV. Methods: Human lung adenocarcinoma epithelial cells (A549) were subcultured, infected with RSV and treated with Nar 30-100 μmol/L. After it was treated for 24 hrs, the expression of MUC5AC at mRNA and protein level in the groups were determined by real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA). The morphological changes of MUC5AC protein were observed with immunofluorescence technology. Results: The expressions of MUC5AC protein and mRNA in all RSV infected groups were significantly higher than that in group C in a dose-dependent manners (all P<0.05). Nar of 30 and 100 μmol/Lsignificantly and dose-dependently decreased RSV-induced secretion of MUC5AC protein in cell supernatant and expression of MUC5AC mRNA (P<0.05), but still higher than that in group C (P<0.05). We also observed that RSV increased MUC5AC protein expression in the cytoplasm of A549 by immunofluorescence technology, and the protein expression decreased after pretreatment with Nar. Conclusion: Naringenin can inhibite the RSV-induced mucous hypersecretion of A549 cell.

Key words： Naringenin RSV mucous hypersecretion MUC5AC

收稿日期: 2015-03-18

基金资助:卫计委国家临床重点专科开放基金资助项目（20130217）。

通讯作者: 张海邻，主任医师，硕士生导师，Email：zhlwz97@hotmail.com。

作者简介: 孟珊珊（1988-），女，山东泰安人，住院医师。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2015/V45/I9/631

[1] Nair H, Nokes DJ, Gessner BD, et al. Global burden of acute lower respiratory infections due to respiratory syncytial virus in young children: a systematic review and meta-analysis[J].Lancet, 2010, 375(9725): 1545-1555.
[2] 林立, 李昌崇. 呼吸道合胞病毒感染发病机制[J]. 中华儿科杂志, 2006, 44(9): 673-675.
[3] Stokes KL, Currier MG, Sakamoto K, et al. The respiratory syncytial virus fusion protein and neutrophils mediate the airway mucin response to pathogenic respiratory syncytial virus infection[J]. J Virol, 2013, 87(18): 10070-10082.
[4] Nadel JA. Mucous hypersecretion and relationship to cough[J]. Pulm Pharmacol Ther, 2013, 26(5): 510-513.
[5] Voynow JA, Rubin BK. Mucins, mucus, and sputum[J].Chest, 2009, 135(2): 505-512.
[6] Nie YC, Wu H, Li PB, et al. Anti-inflammatory effects of naringin in chronic pulmonary neutrophilic inflammation in cigarette smoke-exposed rats[J]. J Med Food, 2012, 15(10):894-900.
[7] Lin BQ, Li PB, Wang YG, et al. The expectorant activity of naringenin[J]. Pulm Pharmacol Ther, 2008, 21(2): 259-263.
[8] Gao S, Li P, Yang H, et al. Antitussive effect of naringin on experimentally induced cough in Guinea pigs[J]. Planta Med, 2011, 77(1): 16-21.
[9] Rose MC, Voynow JA. Respiratory tract mucin genes and mucin glycoproteins in health and disease[J]. Physiol Rev, 2006, 86(1): 245-278.
[10] Miller AL, Strieter RM, Gruber AD, et al. CXCR2 regulates respiratory syncytial virus-induced airway hyperreactivity and mucus overproduction[J]. J Immunol, 2003, 170(6):3348-3356.
[11] 鲍一笑, 董晓艳, 吕婕. 呼吸道合胞病毒诱导气道上皮粘液分泌及调节机制研究[J]. 临床儿科杂志, 2006, 24(1): 25-27.
[12] Lukacs NW, Moore ML, Rudd BD, et al. Differential immune responses and pulmonarypathophysiology are induced by two different strains of respiratory syncytial virus[J]. Am J Pathol, 2006, 169(3): 977-986.
[13] Mata M, Morcillo E, Gimeno C, et al. N-acetyl-L-cysteine (NAC) inhibit mucin synthesis and pro-inflammatory mediators in alveolar type II epithelial cells infected with influenza virus A and B and with respiratory syncytial virus (RSV)[J]. Biochem Pharmacol, 2011, 82(5): 548-555.
[14] Martin L, Chi MH, Luongo C, et al. A chimeric A2 strain of respiratory syncytial virus (RSV) with the fusion protein of RSV strain line 19 exhibits enhanced viral load, mucus, and airway dysfunction[J]. J Virol, 2009, 83(9): 4185-4194.
[15] Shi Y, Dai J, Liu H, et al. Naringenin inhibits allergen-induced airway inflammation and airway responsiveness and inhibits NF-kappaB activity in a murine model of asthma[J].
Can J Physiol Pharmacol, 2009, 87(9): 729-735.
[16] Nie YC, Wu H, Li PB, et al. Naringin attenuates EGF-induced MUC5AC secretion in A549 cells by suppressing the cooperative activities of MAPKs-AP-1 and IKKs-IkappaB-NF-kappaB signaling pathways[J]. Eur J Pharmacol, 2012, 690(1-3): 207-213.