花青素通过Nrf-2/HO-1信号通路调控哮喘气道炎症
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摘要
目的探讨花青素在哮喘小鼠模型中氧化应激的作用以及可能的作用机制。方法50只雌性Balb/c小鼠随机分成对照组、模型组、花青素低剂量组、花青素中剂量组、花青素高剂量组。体外卵清蛋白(OVA)诱导建立哮喘模型,Diff-quik染色后观察细胞总数和各分类细胞数,肺组织切片HE染色观察肺部炎症情况。ELISA方法检测花青素对支气管肺泡灌洗液(BALF)中白细胞介素-4(IL-4)、IL-5和IL-13表达的影响。使用活性氧检测试剂盒通过荧光光谱法定量活性氧(ROS)。Western blot方法检测NF-κB表达水平的变化。Western blot和免疫组化观察Nrf2和HO-1的表达改变。结果与正常组相比,模型组中炎症细胞计数、IL-4、IL-5和IL-13表达升高,ROS含量升高,细胞核内NF-κB p65表达增高(P<0.05);与模型组相比,花青素中、高剂量组中,炎症细胞数、IL-4、IL-5和IL-13的表达下降,ROS含量下降,细胞核内NF-κB p65表达下降,HO-1表达水平和Nrf2核转位明显升高(P<0.05)。结论花青素通过激活Nrf-2/HO-1信号通路调控哮喘小鼠的气道炎症。
Asthma is a phenotypically heterogeneous disorder of the airways, characterized by inflammatory and structural changes. Anthocyanins provide significant protection against many chronic diseases. The objective of this study is to investigate the possible mechanisms of anthocyanins in reducing airway inflammation of asthmatic mice. Diff-quik was used to observe the numbers of the total cells and classified cells; the lung tissue inflammation was observed by HE staining. ELISA was used to detect the levels of interleukin-4(IL-4), IL-5 and IL-13 in BALF.Reactive oxygen species(ROS) was quantified by the reactive oxygen detection kit with Flow cytometer; Western blot was used to detect the level of NF-κB in nucleus; while the expression of Nrf2 and HO-1 were determined by Western blotting and immunohistochemistry. Data showed that anthocyanins could inhibit the airway inflammation of the nuclear transfer of NF-κB and the increase of ROS in the asthmatic mice, whereas improve the nuclear location of Nrf2 and protein levels of HO-1. These results suggested that anthocyanins can regulate the airway inflammation through Nrf2/HO-1 signaling in asthmatic mice.
Asthma is a phenotypically heterogeneous disorder of the airways, characterized by inflammatory and structural changes. Anthocyanins provide significant protection against many chronic diseases. The objective of this study is to investigate the possible mechanisms of anthocyanins in reducing airway inflammation of asthmatic mice. Diff-quik was used to observe the numbers of the total cells and classified cells; the lung tissue inflammation was observed by HE staining. ELISA was used to detect the levels of interleukin-4(IL-4), IL-5 and IL-13 in BALF.Reactive oxygen species(ROS) was quantified by the reactive oxygen detection kit with Flow cytometer; Western blot was used to detect the level of NF-κB in nucleus; while the expression of Nrf2 and HO-1 were determined by Western blotting and immunohistochemistry. Data showed that anthocyanins could inhibit the airway inflammation of the nuclear transfer of NF-κB and the increase of ROS in the asthmatic mice, whereas improve the nuclear location of Nrf2 and protein levels of HO-1. These results suggested that anthocyanins can regulate the airway inflammation through Nrf2/HO-1 signaling in asthmatic mice.
引文
[1]李彬彬,史菲.肿瘤坏死因子超家族成员14在哮喘中的研究进展[J].免疫学杂志,2017,33(7):629-633.
[2]Ma Y,Zhang J,Liu Y,et al.Caffeic acid phenethyl ester alleviates asthma by regulating the airway microenvironment via the ROS-responsive MAPK/Akt pathway[J].Free Radical Biol Med,2016,101:163-175.
[3]Loboda A,Damulewicz M,Pyza E,et al.Role of Nrf2/HO-1 system in development,oxidative stress response and diseases:an evolutionarily conserved mechanism[J].Cell Mol Life Sci,2016,73(17):3221-3247.
[4]Anette K,Lars R,Petter L,Ingvild P,et al.Anthocyanins inhibit nuclear factor-κB activation in monocytes and reduce plasma concentrations of pro-inflammatory mediators in healthy adults[J].J Nutr,2007,137(8):1951-1954.
[5]Park S,Shin W,Seo J,et al.Anthocyanins inhibit airway inflammation and hyperresponsiveness in a murine asthma model[J].Food Chem Toxicol,2007,45(8):1459-1467.
[6]Lee KS,Park HS,Park SJ,et al.An antioxidant modulates expression of receptor activator of NF-kappaB in asthma[J].Exp Mol Med,2006,38(3):217-229.
[7]刘彦群,延光海,秦向征,等.芝麻素通过调节p38和NF-κB激活抑制肥大细胞炎症细胞因子释放[J].免疫学杂志,2014,30(12):1073-1076.
[8]李良昌,金光玉,郑明昱,等.Rottlerin在哮喘气道重塑模型中对PKC-δ/mTOR/NF-κB信号通路的影响[J].免疫学杂志,2013,29(7):588-592.
[9]Gowd V,Jia Z,Chen W.Anthocyanins as promising molecules and dietary bioactive components against diabetes-A review of recent advances[J].Trends Food Sci Technol,2017,68:1-13.
[10]刘英姿,罗有梁,周铁忠.原花青素A-1调节ConA刺激小鼠脾细胞分泌Th1/Th2细胞因子的影响[J].免疫学杂志,2012,28(11):942-945.
[11]周丹阳,曹琦,黄茂,等.原花青素抑制哮喘小鼠气道炎症及气道高反应性[J].南京医科大学学报(自然科学版),2011,31(7):981-985.
[12]Qi S,Xin Y,Guo Y,et al.Ampelopsin reduces endotoxic inflammation via repressing ROS-mediated activation of PI3K/Akt/NF-κB signaling pathways[J].Int Immunopharmacol,2012,12(1):278-287.
[13]Ray PD,Huang B,Tsuji Y.Reactive oxygen species(ROS)homeostasis and redox regulation in cellular signaling[J].Cell Signal,2012,24(5):981-990.
[14]Bellezza I,Giambanco I,Minelli A,et al.Nrf2-Keap1signaling in oxidative and reductive stress[J].BBA Mol Cell Res,2018,1865(5):721-733.
[15]Song Y,Huang L,Yu J.Effects of blueberry anthocyanins on retinal oxidative stress and inflammation in diabetes through Nrf2/HO-1 signaling[J].J Neuroimmunol,2016,301:1-6.
[2]Ma Y,Zhang J,Liu Y,et al.Caffeic acid phenethyl ester alleviates asthma by regulating the airway microenvironment via the ROS-responsive MAPK/Akt pathway[J].Free Radical Biol Med,2016,101:163-175.
[3]Loboda A,Damulewicz M,Pyza E,et al.Role of Nrf2/HO-1 system in development,oxidative stress response and diseases:an evolutionarily conserved mechanism[J].Cell Mol Life Sci,2016,73(17):3221-3247.
[4]Anette K,Lars R,Petter L,Ingvild P,et al.Anthocyanins inhibit nuclear factor-κB activation in monocytes and reduce plasma concentrations of pro-inflammatory mediators in healthy adults[J].J Nutr,2007,137(8):1951-1954.
[5]Park S,Shin W,Seo J,et al.Anthocyanins inhibit airway inflammation and hyperresponsiveness in a murine asthma model[J].Food Chem Toxicol,2007,45(8):1459-1467.
[6]Lee KS,Park HS,Park SJ,et al.An antioxidant modulates expression of receptor activator of NF-kappaB in asthma[J].Exp Mol Med,2006,38(3):217-229.
[7]刘彦群,延光海,秦向征,等.芝麻素通过调节p38和NF-κB激活抑制肥大细胞炎症细胞因子释放[J].免疫学杂志,2014,30(12):1073-1076.
[8]李良昌,金光玉,郑明昱,等.Rottlerin在哮喘气道重塑模型中对PKC-δ/mTOR/NF-κB信号通路的影响[J].免疫学杂志,2013,29(7):588-592.
[9]Gowd V,Jia Z,Chen W.Anthocyanins as promising molecules and dietary bioactive components against diabetes-A review of recent advances[J].Trends Food Sci Technol,2017,68:1-13.
[10]刘英姿,罗有梁,周铁忠.原花青素A-1调节ConA刺激小鼠脾细胞分泌Th1/Th2细胞因子的影响[J].免疫学杂志,2012,28(11):942-945.
[11]周丹阳,曹琦,黄茂,等.原花青素抑制哮喘小鼠气道炎症及气道高反应性[J].南京医科大学学报(自然科学版),2011,31(7):981-985.
[12]Qi S,Xin Y,Guo Y,et al.Ampelopsin reduces endotoxic inflammation via repressing ROS-mediated activation of PI3K/Akt/NF-κB signaling pathways[J].Int Immunopharmacol,2012,12(1):278-287.
[13]Ray PD,Huang B,Tsuji Y.Reactive oxygen species(ROS)homeostasis and redox regulation in cellular signaling[J].Cell Signal,2012,24(5):981-990.
[14]Bellezza I,Giambanco I,Minelli A,et al.Nrf2-Keap1signaling in oxidative and reductive stress[J].BBA Mol Cell Res,2018,1865(5):721-733.
[15]Song Y,Huang L,Yu J.Effects of blueberry anthocyanins on retinal oxidative stress and inflammation in diabetes through Nrf2/HO-1 signaling[J].J Neuroimmunol,2016,301:1-6.
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