| 陶 梅,赵曙光,赵 丽,张 哲,李武良,王晓叶.FABP4对脂多糖诱导Kupffer细胞NF-κB活化和炎症的影响[J].,2019,19(10):1845-1851 |
| FABP4对脂多糖诱导Kupffer细胞NF-κB活化和炎症的影响 |
| The Role of FABP4 on LPS induced Kupffer Cells NF-κB Pathway Activation and Inflammation Response |
| 投稿时间：2018-09-24 修订日期：2018-10-20 |
| DOI：10.13241/j.cnki.pmb.2019.10.009 |
| 中文关键词: NAFLD Kupffer细胞 FABP4 NF-κB 炎症反应 |
| 英文关键词: NAFLD Kupffer cells FABP4 NF-κB Inflammation |
| 基金项目:陕西省社会发展科技攻关项目（2016SF-303）；西安市科技计划项目（2016045SF/YX01(4)） |
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| 中文摘要: |
| 摘要 目的：研究脂肪型脂肪酸结合蛋白（FABP4）对脂多糖（LPS）诱导Kupffer细胞（KCs）NF-κB通路活化和炎症反应的影响。方法：通过梯度离心的方法分离大鼠KCs，按照1×105接种于6孔板，贴壁后饥饿24 h，不同浓度脂多糖（LPS，0、5、10和20 ng/mL）刺激24 h，提取蛋白和RNA，通过Western-Blot检测NF-κB通路蛋白表达变化，利用荧光定量PCR检测IL-1β和IL-6 mRNA表达变化；利用RNAi沉默KCs FABP4表达，通过Western-Blot和荧光定量PCR检测其对LPS诱导NF-κB通路活化的影响；分别利用FABP4细胞因子刺激和慢病毒上调FABP4的表达，通过Western-Blot和荧光定量PCR检测其对KCs NF-κB通路和炎症反应的影响。结果：LPS能够以浓度依赖的方式（0、5、10和20 ng/mL）诱导KCs FABP4 mRNA和蛋白的表达，以20 ng/mL最为明显(P<0.05)；沉默FABP4可以显著减弱LPS（20 ng/mL）诱导的p-p65和p-IκBα的表达，以及炎症细胞因子IL-1β和IL-6的释放(P<0.05)；外源性FABP4（10 ng/mL和20 ng/mL）刺激24h后，能够明显诱导p-p65和p-IκBα的表达，促进炎症因子（IL-1β和IL-6）的合成(P<0.05)；利用慢病毒上调FABP4，可以显著诱导p-p65和p-IκBα的表达以及炎症因子（IL-1β和IL-6）的表达(P<0.05)，而抗氧化剂NAC（10 μM）处理，则显著减弱此效应(P<0.05)。结论：FABP4介导了LPS刺激KCs NF-κB通路的活化和炎症反应。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the role of FABP4 on LPS induced Kupffer cells NF-κB pathway activation and inflammation response. Methods: Liver Kupffer cells were isolated from SD rats using gradient centrifugation. Isolated KCs were inoculated into 6 well cell culture plate with the density of 1×105/mL，then were treated with different concentrations LPS(0, 5, 10, 20 ng/mL) for 24 hours．NF-κB pathway activation was examined through Western-Blot. IL-1β and IL6 expressions were detected through qRT-PCR. Then KCs were treated with LPS for 24 hours after FABP4 knockdown and NF-κB pathway activation was examined. Further, KCs were treated with recombination FABP4 cytokines (10 and 20 ng/mL) or transfected with FABP4 lentivirus and NF-κB pathway activation was examined. Lastly, KCs transfection with FABP4 lentivirus were treated with NAC (10 μM) and then NF-κB pathway activation were examined. Results: Exogenous LPS (0, 5, 10, 20 ng/mL) treatment could significantly promote the expression of FABP4 in Kupffer cells on both mRNA and protein levels in concentration-dependent pattern with most obvious at 20 ng/mL (P<0.05). Knockdown of FABP4 could significantly alleviate LPS induced p65 and IκB phosphorylation and inflammation factors (IL-1β and IL6) expressions in KCs (P<0.05). Exogenous FABP4 cytokines (10 and 20 ng/mL) treatment or transfection with FABP4 lentivirus could significantly promote p65 and IκB phosphorylation with IL-1β and IL6 expressions in KCs (P<0.05). And NAC (10 μM) treatment could significantly alleviate NF-κB pathway activation and inflammation factors expressions in KCs after FABP4 lentivirus transfection (P<0.05). Conclusion: FABP4 is involved in LPS induced Kupffer cells NF-κB pathway activation and inflammation response. |
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