鱼腥草素钠对急性哮喘模型小鼠炎症的影响及机制研究
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摘要
目的观察鱼腥草素钠对急性哮喘模型小鼠的抗炎作用,并探讨作用机制。方法建立卵白蛋白(OVA)诱导小鼠急性哮喘模型,将造模小鼠随机分为模型组,鱼腥草素钠低、高剂量(10、25 mg/kg)组,每组8只,另取正常小鼠8只作为对照组,ig给药2周,对照组与模型组给予等体积生理盐水。采用小鼠肺功能仪器检测小鼠气道高反应性;ELISA法检测血清中特异性OVA-IgE、白细胞介素-4(IL-4)、单核细胞趋化因子-1(MCP-1)浓度;实时荧光定量PCR(qRT-PCR)法检测肺组织中Toll样受体-4(TLR-4)、髓样分化因子88(MyD88)、转铁蛋白6(TRF6)mRNA的表达水平;HE染色后,光镜观察肺组织病理变化。结果与模型组比较,10、25 mg/kg鱼腥草素钠组小鼠在乙酰甲胆碱激发浓度气道高反应值、血清OVE-IgE水平、血清IL-4和MCP浓度均显著降低,具有统计学差异(P<0.05、0.01);与模型组比较,10 mg/kg鱼腥草素钠组肺组织MyD88、TRF6 mRNA表达和25 mg/kg鱼腥草素钠组肺组织TLR-4、MyD88、TRF6 mRNA表达显著降低,具有统计学差异(P<0.05、0.01)。HE染色显示,鱼腥草素钠显著改善模型小鼠的肺组织结构紊乱、肺泡塌陷、支气管周围大量炎症细胞浸润、管腔内有大量分泌物等症状。结论鱼腥草素钠能显著抑制急性哮喘模型小鼠的炎症,机制可能与调节TLR4-NF-κB信号通路有关。
Objective To observe the anti-inflammatory effects of sodium houttuyfonate(SH) on mice with acute asthma and to explore the specific mechanism. Methods OVA-induced acute asthma model in mice was established. The model mice were randomly divided into model group, low and high doses of sodium houttuynin(10, 25 mg/kg) group, 8 mice in each group, and 8 normal mice were taken as control group. After 2 weeks of Ig administration, the control group and model group were given normal saline of equal volume. The airway hyperresponsiveness of mice was measured by pulmonary function instrument, the specific concentrations of OVA-IgE, IL-4 and MCP-1 in serum were detected by ELISA, and the mRNA expression levels of Toll-like receptor-4(TLR-4), myeloid differentiation factor 88(MyD88) and transferrin 6(TRF6) in lung tissue were detected by real-time fluorescence quantitative PCR(qRT-PCR). After HE staining, the pathological changes of lung tissue were observed under light microscope. Results Compared with the model group, the airway hyperresponsiveness, serum OVE-IgE level, serum IL-4 and MCP levels of mice in 10 and 25 mg/kg sodium houttuynin group were significantly decreased(P < 0.05, 0.01); Compared with the model group, the expression of MyD88, TRF6 in lung tissue of 10 mg/kg sodium houttuynin group and TLR-4, MyD88, MyD88, and MCP in lung tissue of 25 mg/kg sodium houttuynin group were significantly decreased(P < 0.05, 0.01). HE staining showed that sodium houttuynin significantly improved the symptoms of pulmonary tissue disorder, alveolar collapse, infiltration of inflammatory cells around the bronchi, and a large number of secretions in the lumen of the model mice. Conclusion Sodium houttuynia significantly inhibited inflammation in an acute asthma mice model and the mechanism may be related to the regulation of the TLR-4-NF-κB signaling pathway.
Objective To observe the anti-inflammatory effects of sodium houttuyfonate(SH) on mice with acute asthma and to explore the specific mechanism. Methods OVA-induced acute asthma model in mice was established. The model mice were randomly divided into model group, low and high doses of sodium houttuynin(10, 25 mg/kg) group, 8 mice in each group, and 8 normal mice were taken as control group. After 2 weeks of Ig administration, the control group and model group were given normal saline of equal volume. The airway hyperresponsiveness of mice was measured by pulmonary function instrument, the specific concentrations of OVA-IgE, IL-4 and MCP-1 in serum were detected by ELISA, and the mRNA expression levels of Toll-like receptor-4(TLR-4), myeloid differentiation factor 88(MyD88) and transferrin 6(TRF6) in lung tissue were detected by real-time fluorescence quantitative PCR(qRT-PCR). After HE staining, the pathological changes of lung tissue were observed under light microscope. Results Compared with the model group, the airway hyperresponsiveness, serum OVE-IgE level, serum IL-4 and MCP levels of mice in 10 and 25 mg/kg sodium houttuynin group were significantly decreased(P < 0.05, 0.01); Compared with the model group, the expression of MyD88, TRF6 in lung tissue of 10 mg/kg sodium houttuynin group and TLR-4, MyD88, MyD88, and MCP in lung tissue of 25 mg/kg sodium houttuynin group were significantly decreased(P < 0.05, 0.01). HE staining showed that sodium houttuynin significantly improved the symptoms of pulmonary tissue disorder, alveolar collapse, infiltration of inflammatory cells around the bronchi, and a large number of secretions in the lumen of the model mice. Conclusion Sodium houttuynia significantly inhibited inflammation in an acute asthma mice model and the mechanism may be related to the regulation of the TLR-4-NF-κB signaling pathway.
引文
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[5] Murray C S, Woodcock A, Langley S J, et al. Secondary prevention of asthma by the use of Inhaled Fluticasone propionate in Wheezy INfants(IFWIN):double-blind,randomised, controlled study[J]. Lancet, 2006, 368(9537):754-762.
[6]陈叶,贺守第,胡智立,等.香菇多糖对卵白蛋白诱导小鼠哮喘模型树突细胞TIM4表达及炎症影响[J].中国临床药理学与治疗学, 2018, 23(9):998-1002.
[7]李斌恺,赖克方,洪燕华,等.支气管哮喘小鼠气道反应性无创检测方法的建立[J].中华哮喘杂志:电子版,2009, 3(4):243-247.
[8] Pan P, Wang Y J, Han L, et al. Effects of sodium houttuyfonate on expression of NF-κB and MCP-1 in membranous glomerulonephritis[J]. J Ethnopharmacol,2010, 131(1):203-209.
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[12] Sugita K, Steer C A, Martinez-Gonzalez I, et al. Type 2innate lymphoid cells disrupt bronchial epithelial barrier integrity by targeting tight junctions through IL-13 in asthmatic patients[J]. J Allergy Clin Immunol, 2018, 141(1):300-310, e11.
[13] Murdoch J R, Lloyd C M. Chronic inflammation and asthma[J]. Mutat Res, 2010, 690(1/2):24-39.
[14] Matsushita H, Ohta S, Shiraishi H, et al. Endotoxin tolerance attenuates airway allergic inflammation in model mice by suppression of the T-cell stimulatory effect of dendritic cells[J]. Int Immunol, 2010, 22(9):739-747.
[15] Medzhitov R, Preston-Hurlburt P, Janeway C A Jr. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity[J]. Nature, 1997, 388(6640):394-397.
[16] Chirico V, Lacquaniti A, Leonardi S, et al. Acute pulmonary exacerbation and lung function decline in patients with cystic fibrosis:high-mobility group box 1(HMGB1)between inflammation and infection[J]. Clin Microbiol Infect, 2015, 21(4):368.e1-e9.
[17] Wang W Q, Hu X Y, Shen P, et al. Sodium houttuyfonate inhibits LPS-induced inflammatory response via suppressing TLR4/NF-κB signaling pathway in bovine mammary epithelial cells[J]. Microb Pathog, 2017, 107:12-16.
[18]赵春贞,姚红伊,王砚,等. TLR4在LPS所致小鼠肺部炎症中的表达及鱼腥草素钠的影响[A].中国药理学会.第九次全国会员代表大会暨全国药理学术会议论文集[C].北京:医药导报杂志社, 2007.
[2]儿童哮喘组,中国疾病预防控制中心环境与健康相关产品安全所.第三次中国城市儿童哮喘流行病学调查[J].中华儿科杂志, 2013, 51(10):729-735.
[3] Guilbert T W, Morgan W J, Zeiger R S, et al. Long-term inhaled corticosteroids in preschool children at high risk for asthma[J]. N Engl J Med, 2006, 354(19):1985-1997.
[4]陈婧,方建国,吴方建,等.鱼腥草抗炎药理作用机制的研究进展[J].中草药, 2014, 45(2):284-289.
[5] Murray C S, Woodcock A, Langley S J, et al. Secondary prevention of asthma by the use of Inhaled Fluticasone propionate in Wheezy INfants(IFWIN):double-blind,randomised, controlled study[J]. Lancet, 2006, 368(9537):754-762.
[6]陈叶,贺守第,胡智立,等.香菇多糖对卵白蛋白诱导小鼠哮喘模型树突细胞TIM4表达及炎症影响[J].中国临床药理学与治疗学, 2018, 23(9):998-1002.
[7]李斌恺,赖克方,洪燕华,等.支气管哮喘小鼠气道反应性无创检测方法的建立[J].中华哮喘杂志:电子版,2009, 3(4):243-247.
[8] Pan P, Wang Y J, Han L, et al. Effects of sodium houttuyfonate on expression of NF-κB and MCP-1 in membranous glomerulonephritis[J]. J Ethnopharmacol,2010, 131(1):203-209.
[9] Hamid Q, Tulic M. Immunobiology of asthma[J]. Annu Rev Physiol, 2009, 71:489-507.
[10] Lumb, Andrew B. Nunn's Applied Respiratory Physiology[M]. Churchill Livingstone:Elsevier, 2010.
[11] ChungK F.Targetingtheinterleukinpathwayinthetreatment ofasthma[J].Lancet,2015,386(9998):1086-1096.
[12] Sugita K, Steer C A, Martinez-Gonzalez I, et al. Type 2innate lymphoid cells disrupt bronchial epithelial barrier integrity by targeting tight junctions through IL-13 in asthmatic patients[J]. J Allergy Clin Immunol, 2018, 141(1):300-310, e11.
[13] Murdoch J R, Lloyd C M. Chronic inflammation and asthma[J]. Mutat Res, 2010, 690(1/2):24-39.
[14] Matsushita H, Ohta S, Shiraishi H, et al. Endotoxin tolerance attenuates airway allergic inflammation in model mice by suppression of the T-cell stimulatory effect of dendritic cells[J]. Int Immunol, 2010, 22(9):739-747.
[15] Medzhitov R, Preston-Hurlburt P, Janeway C A Jr. A human homologue of the Drosophila Toll protein signals activation of adaptive immunity[J]. Nature, 1997, 388(6640):394-397.
[16] Chirico V, Lacquaniti A, Leonardi S, et al. Acute pulmonary exacerbation and lung function decline in patients with cystic fibrosis:high-mobility group box 1(HMGB1)between inflammation and infection[J]. Clin Microbiol Infect, 2015, 21(4):368.e1-e9.
[17] Wang W Q, Hu X Y, Shen P, et al. Sodium houttuyfonate inhibits LPS-induced inflammatory response via suppressing TLR4/NF-κB signaling pathway in bovine mammary epithelial cells[J]. Microb Pathog, 2017, 107:12-16.
[18]赵春贞,姚红伊,王砚,等. TLR4在LPS所致小鼠肺部炎症中的表达及鱼腥草素钠的影响[A].中国药理学会.第九次全国会员代表大会暨全国药理学术会议论文集[C].北京:医药导报杂志社, 2007.