副干酪乳杆菌L9对卵白蛋白诱发的小鼠过敏性气道炎的调节作用
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摘要
过敏性气道炎(哮喘)是一种以炎症细胞浸润,气道高反应性为特征,Th2主导气道炎症性疾病,主要是敏感人群因吸入花粉、尘螨等外界抗原而引起过敏反应。目前常见的治疗方式是糖皮质激素类药物,其副作用大,患者存在类固醇药物抗性等缺点。研究表明特定益生菌菌株具有免疫调节功能,在缓解过敏性气道炎症状等方面有着巨大的潜力。本研究采用卵白蛋白致敏小鼠急性哮喘模型,通过灌胃4.0×109CFU/d和4.0×107CFU/d活菌以及热致死副干酪乳杆菌L9,研究L9对哮喘的改善作用。结果表明:灌胃L9可以显著降低过敏性气道炎小鼠的气道高反应性,降低支气管肺泡灌洗液中嗜酸性粒细胞比例(P<0.05),减轻肺部中小型气道附近炎症细胞的浸润;L9通过降低小鼠血清中的总的IgE(P<0.001)和IgG1(P<0.05),上调血清中IgG2a(P<0.05),纠正小鼠体内的系统性免疫失衡;L9降低支气管肺泡灌洗液中Th2细胞分泌的细胞因子IL-4、IL-5、IL-13的含量(P<0.05),提高Th1细胞分泌的IFN-γ(P<0.05)和Treg细胞相关的TGF-β(P<0.05),纠正小鼠肺部的Th2/Th1免疫失衡。灌胃热致死L9同样可以显著改善哮喘,纠正系统性免疫失衡和Th2/Th1免疫失衡。结论:副干酪乳杆菌L9通过调节Th1/Th2免疫平衡减缓小时过敏性气道炎症状。
Asthma is an obstructive airway disease featuring airway inflammation, air hyperesponssiveness(AHR)and inclined Th2 immune responses. Asthma is a common allergic reaction among antigen-sensitive people stimulated by pollen, dermatophagoid mite or other allergen. Glucocorticoid therapy is the first choice for asthmatics while more and more researchers are looking for some substitutes because of its side effects and steroid resistance among patients. With more and more evidence accumulated, probiotics show a brilliant therapy potential in modulating immune system and alleviating symptoms of allergic asthma. With 20 d acute asthma model of Ovalbumin-sensitized mice applied, effects of both viable and heat-inactivated L9 at oral dose of 4.0×109 CFU/d and 4.0×107 CFU/d were verificated in this study. Allergic mice with intragastric gavage of Lactobacillus paracasei L9 had significantly lower AHR, lower cell counts of eosinophil in BALF(P<0.05), lower level of lung inflammation. L9 considerably decreased total IgE(P<0.001) and IgG1(P<0.05) in serum and increased IgG2 a(P<0.05) so as to balance systematic immune system. With downward levels of IL-4, Il-5, IL-13(P<0.05) and upward levels of IFN-γ(P<0.05) and TGF-β(P<0.05) in Bronchoalveolar lavage fluid(BALF), L9 adjusted biased Th1/Th2 immune system of lung. Dose-effect relation is observed as well. Besides, Heatinactivated Lactobacillus paracasei L9 also modulated asthma with balancing systematic immune system and Th1/Th2 system. Conclusion: Lactobacillus paracasei L9 is able to adjust Th2/Th1 immune system to alleviate inflammation symptoms of allergic asthma.
Asthma is an obstructive airway disease featuring airway inflammation, air hyperesponssiveness(AHR)and inclined Th2 immune responses. Asthma is a common allergic reaction among antigen-sensitive people stimulated by pollen, dermatophagoid mite or other allergen. Glucocorticoid therapy is the first choice for asthmatics while more and more researchers are looking for some substitutes because of its side effects and steroid resistance among patients. With more and more evidence accumulated, probiotics show a brilliant therapy potential in modulating immune system and alleviating symptoms of allergic asthma. With 20 d acute asthma model of Ovalbumin-sensitized mice applied, effects of both viable and heat-inactivated L9 at oral dose of 4.0×109 CFU/d and 4.0×107 CFU/d were verificated in this study. Allergic mice with intragastric gavage of Lactobacillus paracasei L9 had significantly lower AHR, lower cell counts of eosinophil in BALF(P<0.05), lower level of lung inflammation. L9 considerably decreased total IgE(P<0.001) and IgG1(P<0.05) in serum and increased IgG2 a(P<0.05) so as to balance systematic immune system. With downward levels of IL-4, Il-5, IL-13(P<0.05) and upward levels of IFN-γ(P<0.05) and TGF-β(P<0.05) in Bronchoalveolar lavage fluid(BALF), L9 adjusted biased Th1/Th2 immune system of lung. Dose-effect relation is observed as well. Besides, Heatinactivated Lactobacillus paracasei L9 also modulated asthma with balancing systematic immune system and Th1/Th2 system. Conclusion: Lactobacillus paracasei L9 is able to adjust Th2/Th1 immune system to alleviate inflammation symptoms of allergic asthma.
引文
[1] BOCHNER B S, BUSSE W W. Allergy and asthma[J]. Journal of Allergy and Clinical Immunology,2005, 115(5):953-959.
[2] BRAMAN S S. The global burden of asthma[J].Chest Journal, 2006, 130(1_suppl):4S-12S.
[3] BEASLEY R. Worldwide variation in prevalence of symptoms of asthma, allergic rhinoconjunctivitis,and atopic eczema:ISAA C[J]. The Lancet, 1998,351(9111):1225-32.
[4] SAGAR S, MORGAN M E, CHEN S, et al. Bifidobacterium breve and Lactobacillus rhamnosus treatment is as effective as budesonide at reducing inflammation in a murine model for chronic asthma[J]. Respiratory Research, 2014, 15(1):1.
[5] MCGUIRK P, MILLS K H. Pathogen-specific regulatory T cells provoke a shift in the Th1/Th2paradigm in immunity to infectious diseases[J].Trends in Immunology, 2002, 23(9):450-455.
[6] YAMANAKA T, HELGELAND L, FARSTAD I N,et al. Microbial colonization drives lymphocyte accumulation and differentiation in the follicle-associated epithelium of Peyer’s patches[J]. The Journal of Immunology. 2003, 170(2):816-822.
[7] VON DER WEID T, BULLIARD C, SCHIFFRIN E J. Induction by a lactic acid bacterium of a population of CD4+T cells with low proliferative capacity that produce transforming growth factorβand interleukin-10[J]. Clinical and Diagnostic Laboratory Immunology, 2001, 8(4):695-701.
[8] CHRISTENSEN H R, FR覫KI覷R H, PESTKA J J.Lactobacilli differentially modulate expression of cytokines and maturation surface markers in murine dendritic cells[J]. The Journal of Immunology. 2002,168(1):171-178.
[9] FUJIWARA D, INOUE S, WAKABAYASHI H, et al. The anti-allergic effects of lactic acid bacteria are strain dependent and mediated by effects on both Th1/Th2 cytokine expression and balance[J].International archives of allergy and immunology,2004, 135(3):205-215.
[10] SEGAWA S, NAKAKITA Y, TAKATA Y, et al.Effect of oral administration of heat-killed Lactobacillus brevis SBC8803 on total and ovalbuminspecific immunoglobulin E production through the improvement of Th1/Th2 balance[J]. International Journal of Food Microbiology, 2008, 121(1):1-10.
[11] SUNADA Y, NAKAMURA S, KAMEI C. Effects of Lactobacillus acidophilus strain L-55 on experimental allergic rhinitis in BALB/c mice[J]. Biological and Pharmaceutical Bulletin, 2007, 30(11):2163-2166.
[12] SHIDA K, TAKAHASHI R, IWADATE E, et al.Lactobacillus casei strain Shirota suppresses serum immunoglobulin E and immunoglobulin G1 responses and systemic anaphylaxis in a food allergy model[J]. Clinical&Experimental Allergy, 2002, 32(4):563-570.
[13] VAN OVERTVELT L, MOUSSU H, HORIOT S, et al. Lactic acid bacteria as adjuvants for sublingual allergy vaccines[J]. Vaccine, 2010, 28(17):2986-2992.
[14] ASHRAF R, SHAH N P. Immune system stimulation by probiotic microorganisms[J]. Critical Reviews in Food Science and Nutrition, 2014, 54(7):938-956.
[15] JANG S O, KIM H J, KIM Y J, et al. Asthma prevention by Lactobacillus rhamnosus in a mouse model is associated with CD4+CD25+Foxp3+T cells[J]. Allergy, Asthma&Immunology Research, 2012,4(3):150-156.
[16] PELLATON C, NUTTEN S, THIERRY A C, et al.Intragastric and intranasal administration of Lactobacillus paracasei NCC2461 modulates allergic airway inflammation in mice[J]. International Journal of Inflammation, 2012, 2012:1-8
[17] HUANG W K, CAI J H, KORIUI M, et al. Antiasthmatic of Lactobacillus paracasei subsp. Paracasei NTU101 in OVA-induced asthmatic mice[J]. The FASEB Journal, 2013, 27(1 Supplement):1057.
[18] LIU Y W, LIAO T W, CHEN Y H, et al. Oral administration of heat-inactivated Lactobacillus plantarum K37 modulated airway hyperresponsiveness in ovalbumin-sensitized BALB/c mice[J]. PloS one, 2014,9(6):e100105.
[19]何敏,王婷,张红萍,等.益生菌预防和治疗支气管哮喘的系统评价[J].中国循证医学杂志, 2012,12(4):460-469
[20] YANG J, REN F, ZHANG H, et al. Induction of regulatory dendritic cells by Lactobacillus paracasei L9 prevents allergic sensitization to bovineβ-lactoglobulin in mice[J]. J Microbiol Biotechnol, 2015,25:1687-1696.
[21] ZHU Y, ZHU J, ZHAO L,et al. Effect of oral administration of Lactobacillus paracasei L9 on mouse systemic immunity and the immune response in the intestine[J]. Archives of Biological Sciences, 2016,68(2):311-318.
[22]郝明明.生命早期益生菌干预对预防过敏性疾病的研究[D].石家庄:河北医科大学, 2013.
[23] SUDO N, SAWAMURA S A, TANAKA K,et al.The requirement of intestinal bacterial flora for the development of an IgE production system fully susceptible to oral tolerance induction[J]. The Journal of Immunology. 1997, 159(4):1739-1745.
[24] GALDEANO C M, PERDIGON G. The probiotic bacterium Lactobacillus casei induces activation of the gut mucosal immune system through innate immunity[J]. Clinical and Vaccine Immunology, 2006,13(2):219-226.
[25] VIEIRA A T, TEIXEIRA M M, MARTINS F d S.The role of probiotics and prebiotics in inducing gut immunity[J]. Frontiers in Immunology, 2013, 12(4):445.
[26] FREI R, AKDIS M, O'MAHONY L. Prebiotics,probiotics, synbiotics, and the immune system:ex perimental data and clinical evidence[J]. Current Opinion in Gastroenterology, 2015, 31(2):153-158.
[27] FORSYTHE P. Probiotics and lung immune respons es[J]. Annals of the American Thoracic Society, 2014,11(Supplement 1):S33-S37.
[28] JOHNSON J L, JONES M B, COBB B A. Bacterial capsular polysaccharide prevents the onset of asthma through T cell activation[J]. Glycobiology, 2015, 25(4):368-375.
[29] TSUCHIYA K, SIDDIQUI S, RISSE P A, et al.The presence of LPS in OVA inhalations affects airway inflammation and AHR but not remodeling in a rodent model of asthma[J]. American Journal of Phys iology-Lung Cellular and Molecular Physiology,2012, 303(1):L54-L63.
[2] BRAMAN S S. The global burden of asthma[J].Chest Journal, 2006, 130(1_suppl):4S-12S.
[3] BEASLEY R. Worldwide variation in prevalence of symptoms of asthma, allergic rhinoconjunctivitis,and atopic eczema:ISAA C[J]. The Lancet, 1998,351(9111):1225-32.
[4] SAGAR S, MORGAN M E, CHEN S, et al. Bifidobacterium breve and Lactobacillus rhamnosus treatment is as effective as budesonide at reducing inflammation in a murine model for chronic asthma[J]. Respiratory Research, 2014, 15(1):1.
[5] MCGUIRK P, MILLS K H. Pathogen-specific regulatory T cells provoke a shift in the Th1/Th2paradigm in immunity to infectious diseases[J].Trends in Immunology, 2002, 23(9):450-455.
[6] YAMANAKA T, HELGELAND L, FARSTAD I N,et al. Microbial colonization drives lymphocyte accumulation and differentiation in the follicle-associated epithelium of Peyer’s patches[J]. The Journal of Immunology. 2003, 170(2):816-822.
[7] VON DER WEID T, BULLIARD C, SCHIFFRIN E J. Induction by a lactic acid bacterium of a population of CD4+T cells with low proliferative capacity that produce transforming growth factorβand interleukin-10[J]. Clinical and Diagnostic Laboratory Immunology, 2001, 8(4):695-701.
[8] CHRISTENSEN H R, FR覫KI覷R H, PESTKA J J.Lactobacilli differentially modulate expression of cytokines and maturation surface markers in murine dendritic cells[J]. The Journal of Immunology. 2002,168(1):171-178.
[9] FUJIWARA D, INOUE S, WAKABAYASHI H, et al. The anti-allergic effects of lactic acid bacteria are strain dependent and mediated by effects on both Th1/Th2 cytokine expression and balance[J].International archives of allergy and immunology,2004, 135(3):205-215.
[10] SEGAWA S, NAKAKITA Y, TAKATA Y, et al.Effect of oral administration of heat-killed Lactobacillus brevis SBC8803 on total and ovalbuminspecific immunoglobulin E production through the improvement of Th1/Th2 balance[J]. International Journal of Food Microbiology, 2008, 121(1):1-10.
[11] SUNADA Y, NAKAMURA S, KAMEI C. Effects of Lactobacillus acidophilus strain L-55 on experimental allergic rhinitis in BALB/c mice[J]. Biological and Pharmaceutical Bulletin, 2007, 30(11):2163-2166.
[12] SHIDA K, TAKAHASHI R, IWADATE E, et al.Lactobacillus casei strain Shirota suppresses serum immunoglobulin E and immunoglobulin G1 responses and systemic anaphylaxis in a food allergy model[J]. Clinical&Experimental Allergy, 2002, 32(4):563-570.
[13] VAN OVERTVELT L, MOUSSU H, HORIOT S, et al. Lactic acid bacteria as adjuvants for sublingual allergy vaccines[J]. Vaccine, 2010, 28(17):2986-2992.
[14] ASHRAF R, SHAH N P. Immune system stimulation by probiotic microorganisms[J]. Critical Reviews in Food Science and Nutrition, 2014, 54(7):938-956.
[15] JANG S O, KIM H J, KIM Y J, et al. Asthma prevention by Lactobacillus rhamnosus in a mouse model is associated with CD4+CD25+Foxp3+T cells[J]. Allergy, Asthma&Immunology Research, 2012,4(3):150-156.
[16] PELLATON C, NUTTEN S, THIERRY A C, et al.Intragastric and intranasal administration of Lactobacillus paracasei NCC2461 modulates allergic airway inflammation in mice[J]. International Journal of Inflammation, 2012, 2012:1-8
[17] HUANG W K, CAI J H, KORIUI M, et al. Antiasthmatic of Lactobacillus paracasei subsp. Paracasei NTU101 in OVA-induced asthmatic mice[J]. The FASEB Journal, 2013, 27(1 Supplement):1057.
[18] LIU Y W, LIAO T W, CHEN Y H, et al. Oral administration of heat-inactivated Lactobacillus plantarum K37 modulated airway hyperresponsiveness in ovalbumin-sensitized BALB/c mice[J]. PloS one, 2014,9(6):e100105.
[19]何敏,王婷,张红萍,等.益生菌预防和治疗支气管哮喘的系统评价[J].中国循证医学杂志, 2012,12(4):460-469
[20] YANG J, REN F, ZHANG H, et al. Induction of regulatory dendritic cells by Lactobacillus paracasei L9 prevents allergic sensitization to bovineβ-lactoglobulin in mice[J]. J Microbiol Biotechnol, 2015,25:1687-1696.
[21] ZHU Y, ZHU J, ZHAO L,et al. Effect of oral administration of Lactobacillus paracasei L9 on mouse systemic immunity and the immune response in the intestine[J]. Archives of Biological Sciences, 2016,68(2):311-318.
[22]郝明明.生命早期益生菌干预对预防过敏性疾病的研究[D].石家庄:河北医科大学, 2013.
[23] SUDO N, SAWAMURA S A, TANAKA K,et al.The requirement of intestinal bacterial flora for the development of an IgE production system fully susceptible to oral tolerance induction[J]. The Journal of Immunology. 1997, 159(4):1739-1745.
[24] GALDEANO C M, PERDIGON G. The probiotic bacterium Lactobacillus casei induces activation of the gut mucosal immune system through innate immunity[J]. Clinical and Vaccine Immunology, 2006,13(2):219-226.
[25] VIEIRA A T, TEIXEIRA M M, MARTINS F d S.The role of probiotics and prebiotics in inducing gut immunity[J]. Frontiers in Immunology, 2013, 12(4):445.
[26] FREI R, AKDIS M, O'MAHONY L. Prebiotics,probiotics, synbiotics, and the immune system:ex perimental data and clinical evidence[J]. Current Opinion in Gastroenterology, 2015, 31(2):153-158.
[27] FORSYTHE P. Probiotics and lung immune respons es[J]. Annals of the American Thoracic Society, 2014,11(Supplement 1):S33-S37.
[28] JOHNSON J L, JONES M B, COBB B A. Bacterial capsular polysaccharide prevents the onset of asthma through T cell activation[J]. Glycobiology, 2015, 25(4):368-375.
[29] TSUCHIYA K, SIDDIQUI S, RISSE P A, et al.The presence of LPS in OVA inhalations affects airway inflammation and AHR but not remodeling in a rodent model of asthma[J]. American Journal of Phys iology-Lung Cellular and Molecular Physiology,2012, 303(1):L54-L63.