| 呼吸道合胞病毒感染后IL-17、hCLCA1表达变化及罗格列酮的抑制效应 |
| 投稿时间：2016-10-17 修订日期：2016-11-20 点此下载全文 |
| 引用本文：杨康康,董琳,丁洁.呼吸道合胞病毒感染后IL-17、hCLCA1表达变化及罗格列酮的抑制效应[J].医学研究杂志,2017,46(9):107-111 |
| DOI：
10.11969/j.issn.1673-548X.2017.09.029 |
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| 基金项目:浙江省温州市科技局公益技术研究科技合作项目（H20140002） |
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| 中文摘要:目的 研究呼吸道合胞病毒（RSV）感染A549细胞后过氧化物酶体增殖物激活受体γ（PPARγ）、IL-17及人钙激活氯离子通道辅助蛋白1（hCLCA1） mRNA和蛋白表达变化及罗格列酮和IL-17抗体的干预作用。方法 将传代细胞随机分6组：罗格列酮/RSV组、罗格列酮/GW9662/RSV组、DMSO/RSV组、IL-17抗体/RSV组、GW9662/RSV组、细胞对照组。DMSO/RSV组予0.02% DMSO预处理0.5h，罗格列酮/RSV组予罗格列酮（10μmol/L）预处理0.5h，罗格列酮/GW9662/RSV组在应用罗格列酮前先以GW9662（10μmol/L）预处理0.5h，IL-17抗体/RSV组在RSV感染前予IL-17抗体预处理2h。各组培养6、12、24h收获细胞及上清液待测。应用实时荧光定量RT-PCR检测PPARγ、IL-17、hCLCA1 mRNA表达，Western blot法检测hCLCA1蛋白水平，ELISA检测IL-17蛋白表达。结果 与细胞对照组相比，DMSO/RSV组PPARγ、IL-17、hCLCA1 mRNA和蛋白表达在3个时间点均明显升高，随时间增加表达量增加，24h达高峰，与12h比较，差异有统计学意义（P均< 0.05）；在同一时间点上罗格列酮/RSV组、IL-17抗体/RSV组IL-17、hCLCA1mRNA和蛋白均明显低于DMSO/RSV组，与罗格列酮/GW9662/RSV组相比，罗格列酮/RSV组IL-17、hCLCA1mRNA和蛋白表达均明显降低。罗格列酮及IL-17抗体对IL-17 mRNA和蛋白的抑制作用在干预后6h最强，与12、24h相比较，差异有统计学意义（P<0.05）。结论 罗格列酮可抑制RSV感染后IL-17、hCLCA1mRNA及蛋白表达增加，机制可能为上调PPARγ基因的表达，使PPARγ活性增强，在转录水平下调IL-17表达，从而抑制气道黏液高分泌。 |
| 中文关键词:呼吸道合胞病毒 IL-17 hCLCA1 A549细胞 |
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| Changes of IL-17and HCLCA1 Induced by RSV Infection and the Inhibition Effect of Rosiglitazone |
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| Abstract:Objective To observe the expressions of peroxisome proliferator activatedγ (PPARγ), IL-17 and calcium activated family member 1(hCLCA1) mRNA and protein in A549 cell infected with respiratory syncytial virus(RSV),and to explore the effect of rosiglitazone and anti-IL-17 Ab. Methods A549 cells were seeded in 6-well culture plates and randomly divided into six groups. DMSO/RSV group received medium containing 0.02% DMSO 30min prior to RSV infection. Rosiglitazone/RSV group received rosiglitazone (10μmol/L) 30 min prior to RSV infection. GW9662/rosiglitazone/RSV group was first exposed to GW9662(10μmol/L)for 30 min, prior to rosiglitazone and RSV infection. Anti-IL-17 Ab (0.15μmol/L) was administered to anti-IL-17 Ab/RSV group 2 hour prior to RSV infection.Supernatants were harvested at 6h,12h and 24h post-RSV infection, retrospectively. The expression of PPARγ, IL-17,hCLCA1 mRNA were measured by real-time quantitative PCR. Protein levels of hCLCA1 and IL-17 were measured by Western blot and ELISA, retrospectively. Results The levels of PPARγ, IL-17and hCLCA1 mRNA and protein were higher in DMSO/RSV group than those in cell control group (P<0.05) at three time points. The expression of IL-17,hCLCA1 mRNA and protein were lower both in rosiglitazone/RSV group and anti-IL-17 Ab/RSV group than those in DMSO/RSV group at the same time (P<0.05). While, both IL-17,hCLCA1 protein and mRNA expression in GW9662/Rosiglitazone/RSV group were similar to that in DMSO/RSV group (P<0.05). Compared with that at 12h and 24h,rosiglitazone and anti-IL-17 Ab had the strongest inhibition effect onthe expression of IL-17 mRNA and protein at 6h (P<0.05). Conclusion Rosiglitazone can inhibit RSV-induced expression of IL-17, hCLCA1 and may inhibit airway mucus hypersecretion by increasing the activity of PPARγ. |
| keywords:Respiratory syncytial virus IL-17 hCLCA1 A549cell |
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