汉黄芩素对急性肺损伤小鼠的保护作用
|
摘要
目的:研究汉黄芩素(WOG)对脂多糖(LPS)诱导的急性肺损伤(ALI)小鼠的治疗效果及其可能机制。方法:选用C57/BL6小鼠,采用LPS(1μg/kg)气管内灌注的方式构建ALI模型。实验分为3组:空白对照组(Control组)、LPS诱导的ALI组(LPS组)、WOG治疗组(WOG组)。随后分别采用流式细胞术及ELISA法检测肺泡灌洗液(BALF)中免疫细胞(中性粒细胞、巨噬细胞)含量变化及炎性细胞因子分泌情况。在此基础上,采集各实验组小鼠的肺组织进行HE染色。最后,通过对LPS下游的常见通路,即NF-κB信号通路分析,探究WOG对ALI小鼠的保护作用的分子机制。结果:WOG可以显著降低肺组织的湿重/干重比(P<0.05),减轻肺组织中炎性细胞的浸润,抑制TNF-α、IL-1β、IL-6和IL-10等炎性细胞因子的产生(P<0.01)。Westernblot结果显示,WOG可以显著抑制NF-κB信号通路表达。结论:WOG对LPS诱导的ALI小鼠具有保护作用,其机制与NF-κB信号通路的抑制相关。
Objective: To explore the therapeutic effect of wogonin on mice with LPS-induced acute lung injury and its possible mechanism. Methods: The acute lung injury model was constructed by LPS(1 μg/kg) endotracheal instillation using C57/BL6 mice. The experiment was composed of 3 groups: control group, LPSinduced acute lung injury group(LPS group), and wogonin treatment group(WOG group). Flow cytometry and ELISA were used to detect the content of immune cells(neutrophils and macrophages) and the secretion of inflammatory cytokines in bronchoalveolar lavage fluid(BALF). Lung tissues of mice in each experimental group were also collected for hematoxylin-eosin(HE) staining. Finally, the molecular mechanism of the protective effect of wogonin on acute lung injury mice was investigated by analyzing the common downstream pathway of LPS, namely NF-κB signaling pathway. Results: Wogonin can significantly reduce the wet-to-dry(W/D) ratio of lung tissue(P<0.05), the infiltration of inflammatory cells in lung tissue, and the production of TNF-α, IL-1β, IL-6 and IL-10 inflammatory cytokine(P<0.01). The results of Western blot showed that wogonin can significantly inhibit the NF-κB signaling pathway. Conclusion: Wogonin possesses potential protection against LPS-induced acute lung injury through its anti-inflammatory property, and its potential mechanism is associated with NF-κB signaling pathways.
Objective: To explore the therapeutic effect of wogonin on mice with LPS-induced acute lung injury and its possible mechanism. Methods: The acute lung injury model was constructed by LPS(1 μg/kg) endotracheal instillation using C57/BL6 mice. The experiment was composed of 3 groups: control group, LPSinduced acute lung injury group(LPS group), and wogonin treatment group(WOG group). Flow cytometry and ELISA were used to detect the content of immune cells(neutrophils and macrophages) and the secretion of inflammatory cytokines in bronchoalveolar lavage fluid(BALF). Lung tissues of mice in each experimental group were also collected for hematoxylin-eosin(HE) staining. Finally, the molecular mechanism of the protective effect of wogonin on acute lung injury mice was investigated by analyzing the common downstream pathway of LPS, namely NF-κB signaling pathway. Results: Wogonin can significantly reduce the wet-to-dry(W/D) ratio of lung tissue(P<0.05), the infiltration of inflammatory cells in lung tissue, and the production of TNF-α, IL-1β, IL-6 and IL-10 inflammatory cytokine(P<0.01). The results of Western blot showed that wogonin can significantly inhibit the NF-κB signaling pathway. Conclusion: Wogonin possesses potential protection against LPS-induced acute lung injury through its anti-inflammatory property, and its potential mechanism is associated with NF-κB signaling pathways.
引文
[1]MUTLU G M,BUDINGER G R.Incidence and outcomes of acute lung injury[J].N Engl J Med,2006,354(4):416-417.
[2]LIM B O.Efficacy of wogonin in the production of immunoglobulins and cytokines by mesenteric lymph node lymphocytes in mouse colitis induced with dextran sulfate sodium[J].Biosci Biotechnol Biochem,2004,68(12):2505-2511.
[3]CHI Y S,LIM H,PARK H,et al.Effects of wogonin,a plant flavone from Scutellaria radix,on skin inflammation:in vivo regulation of inflammation-associated gene expression[J].Biochem Pharmacol,2003,66(7):1271-1278.
[4]CHEN Y C,SHEN S C,CHEN L G,et al.Wogonin,baicalin,and baicalein inhibition of inducible nitric oxide synthase and cyclooxygenase-2 gene expressions induced by nitric oxide synthase inhibitors and lipopolysaccharide[J].Biochem Pharmacol,2001,61(11):1417-1427.
[5]FERGUSON N D,FRUTOS-VIVAR F,ESTEBAN A,et al.Acute respiratory distress syndrome:underrecognition by clinicians and diagnostic accuracy of three clinical definitions[J].Crit Care Med,2005,33(10):2228-2234.
[6]RUBENFELD G D,CALDWELL E,PEABODY E,et al.Incidence and outcomes of acute lung injury[J].N Engl JMed,2005,353(16):1685-1693.
[7]WARE L B,MATTHAY M A.The acute respiratory distress syndrome[J].N Engl J Med,2000,342(18):1334-1349.
[8]MCLEISH K R,KLEIN J B,LEDERER E D,et al.Azotemia,TNF alpha,and LPS prime the human neutrophil oxidative burst by distinct mechanisms[J].Kidney Int,1996,50(2):407-416.
[9]张佳莹,魏苗苗,初晓,等.甘草黄酮对小鼠急性肺损伤保护机制的研究[J].中国农学通报,2012,28(8):56-62.
[10]徐昉,杨远征,刘琼.姜黄素对脓毒症小鼠急性肺损伤的保护作用及对细胞间黏附分子-1和肿瘤坏死因子-α表达的影响[J].中国生物制品学杂志,2011,24(1):75-78.
[11]CHEN C C,HUNG T H,WANG Y H,et al.Wogonin improves histological and functional outcomes,and reduces activation of TLR4/NF-kappaB signaling after experimental traumatic brain injury[J].PLoS One,2012,7(1):e30294.
[12]YANG R,YANG L,SHEN X,et al.Suppression of NF-kappaB pathway by crocetin contributes to attenuation of lipopolysaccharide-induced acute lung injury in mice[J].Eur JPharmacol,2012,674(2-3):391-396.
[13]FAN J,YE R D,MALIK A.Transcriptional mechanisms of acute lung injury[J].Am J Physiol Lung Cell Mol Physiol,2001,281(5):L1037-1050.
[14]BHATIA M,MOOCHHALA S.Role of inflammatory mediators in the pathophysiology of acute respiratory distress syndrome[J].J Pathol,2004,202(2):145-156.
[15]KOLB M,MARGETTS P J,ANTHONY D C,et al.Transient expression of IL-1beta induces acute lung injury and chronic repair leading to pulmonary fibrosis[J].J Clin Invest,2001,107(12):1529-1536.
[16]IANARO A M,TERSIGNI F,D’ACQUIST O.New insight in LPS antagonist[J].Mini Rev Med Chem,2009,9(3):306-317.
[2]LIM B O.Efficacy of wogonin in the production of immunoglobulins and cytokines by mesenteric lymph node lymphocytes in mouse colitis induced with dextran sulfate sodium[J].Biosci Biotechnol Biochem,2004,68(12):2505-2511.
[3]CHI Y S,LIM H,PARK H,et al.Effects of wogonin,a plant flavone from Scutellaria radix,on skin inflammation:in vivo regulation of inflammation-associated gene expression[J].Biochem Pharmacol,2003,66(7):1271-1278.
[4]CHEN Y C,SHEN S C,CHEN L G,et al.Wogonin,baicalin,and baicalein inhibition of inducible nitric oxide synthase and cyclooxygenase-2 gene expressions induced by nitric oxide synthase inhibitors and lipopolysaccharide[J].Biochem Pharmacol,2001,61(11):1417-1427.
[5]FERGUSON N D,FRUTOS-VIVAR F,ESTEBAN A,et al.Acute respiratory distress syndrome:underrecognition by clinicians and diagnostic accuracy of three clinical definitions[J].Crit Care Med,2005,33(10):2228-2234.
[6]RUBENFELD G D,CALDWELL E,PEABODY E,et al.Incidence and outcomes of acute lung injury[J].N Engl JMed,2005,353(16):1685-1693.
[7]WARE L B,MATTHAY M A.The acute respiratory distress syndrome[J].N Engl J Med,2000,342(18):1334-1349.
[8]MCLEISH K R,KLEIN J B,LEDERER E D,et al.Azotemia,TNF alpha,and LPS prime the human neutrophil oxidative burst by distinct mechanisms[J].Kidney Int,1996,50(2):407-416.
[9]张佳莹,魏苗苗,初晓,等.甘草黄酮对小鼠急性肺损伤保护机制的研究[J].中国农学通报,2012,28(8):56-62.
[10]徐昉,杨远征,刘琼.姜黄素对脓毒症小鼠急性肺损伤的保护作用及对细胞间黏附分子-1和肿瘤坏死因子-α表达的影响[J].中国生物制品学杂志,2011,24(1):75-78.
[11]CHEN C C,HUNG T H,WANG Y H,et al.Wogonin improves histological and functional outcomes,and reduces activation of TLR4/NF-kappaB signaling after experimental traumatic brain injury[J].PLoS One,2012,7(1):e30294.
[12]YANG R,YANG L,SHEN X,et al.Suppression of NF-kappaB pathway by crocetin contributes to attenuation of lipopolysaccharide-induced acute lung injury in mice[J].Eur JPharmacol,2012,674(2-3):391-396.
[13]FAN J,YE R D,MALIK A.Transcriptional mechanisms of acute lung injury[J].Am J Physiol Lung Cell Mol Physiol,2001,281(5):L1037-1050.
[14]BHATIA M,MOOCHHALA S.Role of inflammatory mediators in the pathophysiology of acute respiratory distress syndrome[J].J Pathol,2004,202(2):145-156.
[15]KOLB M,MARGETTS P J,ANTHONY D C,et al.Transient expression of IL-1beta induces acute lung injury and chronic repair leading to pulmonary fibrosis[J].J Clin Invest,2001,107(12):1529-1536.
[16]IANARO A M,TERSIGNI F,D’ACQUIST O.New insight in LPS antagonist[J].Mini Rev Med Chem,2009,9(3):306-317.