表没食子儿茶素没食子酸酯对肥胖型哮喘小鼠气道炎症和Treg/Th17免疫平衡的影响
|
摘要
目的:探讨表没食子儿茶素没食子酸酯(EGCG)对肥胖型哮喘小鼠调节性T淋巴细胞/辅助性T淋巴细胞17 (Treg/Th17)免疫失衡的调节作用,为EGCG治疗肥胖型哮喘提供依据。方法:40只C57BL/6J小鼠分为正常对照组(采用正常饲料喂养)、肥胖组(采用高脂饲料喂养)、肥胖型哮喘组[采用高脂饲料喂养的同时给予卵白蛋白(OVA)致敏和激发,制备肥胖型哮喘模型],EGCG干预组(在OVA激发前1h给予20mg·kg~(-1) EGCG腹腔注射)和地塞米松干预组(在OVA激发前1h给予2mg·kg~(-1)地塞米松腹腔注射),每组8只。采用无创肺功能仪测定各组小鼠的增强呼气间歇(Penh)值;HE染色观察各组小鼠肺组织病理形态表现,HE染色切片半定量方法进行小鼠气道炎症评分;ELISA法检测各组小鼠血清中脂联素、瘦素和支气管肺泡灌洗液(BALF)中白细胞介素10 (IL~(-1)0)、白细胞介素17A (IL~(-1)7A)水平;流式细胞术检测各组小鼠脾组织中Th17和Treg百分比。结果:肥胖组小鼠体质量高于正常对照组(P<0.05),为正常对照组小鼠平均体质量的1.67倍。与正常对照组比较,肥胖型哮喘组小鼠Penh值和气道炎症评分均升高(P<0.05),血清中瘦素水平升高(P<0.05),BALF中IL~(-1)7A水平升高(P<0.05),IL~(-1)0水平降低(P<0.05),脾组织中Th17百分比升高(P<0.05),Treg百分比降低(P<0.05)。与肥胖型哮喘组比较,EGCG干预组小鼠Penh值和气道炎症评分均降低(P<0.05),血清中瘦素水平降低(P<0.05),BALF中IL~(-1)7A水平降低(P<0.05),BALF中IL~(-1)0水平升高(P<0.05),脾组织中Th17百分比降低(P<0.05),脾组织中Treg百分比升高(P<0.05)。结论:在肥胖型哮喘小鼠中存在Treg/Th17免疫失衡,EGCG能够抑制肥胖型哮喘小鼠的气道炎症及气道高反应性,能够改善Treg/Th17免疫失衡。
Objective:To investigate the regulatory effect of epigallocatechin-3-gallate(EGCG)on the immune imbalance of regulatory T lymphocytes/helper T lymphocytes 17(Treg/Th17)in the mice with obese asthma,and to provide the basis for the treatment of obese asthma by EGCG.Methods:A total of 40 C57BL/6J mice were randomly divided into normal control group(fed with chow diet),obese group(fed with high fat diet),obese asthma group [fed with high fat diet and sensitized and challenged with ovalbumin(ova)],EGCG intervention group(20 mg·kg~(-1) EGCG was administered intraperitoneally 1 hbefore challenge of OVA)and dexamethasone intervention group(2 mg·kg~(-1) dexamethasone was administered intraperitoneally 1 hbefore challenge of OVA),and there were 8 mice in each group.The edhanced pause(Penh)values of the mice in various groups were measured with non-invasive lung function instrument;the pathomorphology of lung tissue of the mice in various groups were observed by HE staining,and the inflammation scores were evaluated by semi-quantitative method for HE staining section;the levels of interleukin~(-1)0(IL~(-1)0)and interleukin~(-1)7 A(IL~(-1)7 A)in bronchoalveolar lavage fluid(BALF)and adiponectin and leptin in serum of the mice in various groups were detected by ELISA method,and the percentages of Treg and Th17 in spleen tissue of the mice in various groups were detected by flow cytometery.Results:Compared with normal control group,the body weight of the mice in obese group was increased(P<0.05);it was 1.67 times the average weight of the mice in normal control group.Compared with normal control group,the Penh value and inflammation score of the mice in obese asthma group were increased(P<0.05);the leptin level in serum of the mice in obese asthma group was increased(P<0.05);the IL~(-1)7 level in BALF of the mice in obese asthma group was increased(P<0.05)and IL~(-1)0 level in BALF of the mice in obese asthma group was decreased(P<0.05);the percentage of Th17 in spleen tissue of the mice in obese asthma group was increased(P<0.05)and the percentage of Treg in spleen tissue of the mice in obese asthma group was decreased(P<0.05).Compared with obese asthma group,the Penh value and inflammation score of the mice in EGCG intervention group were decreased(P<0.05);the leptin level in serum of the mice in EGCG intervention group was decreased(P<0.05);the IL~(-1)7 level in BALF of the mice in EGCG intervention group was decreased(P<0.05)and IL~(-1)0 level in BALF of the mice in EGCG intervention group was increased(P<0.05);the percentage of Th17 in spleen tissue of the mice in EGCG intervention group was decreased(P<0.05)and the percentage of Treg in spleen tissue of the mice in EGCG intervention group was increased(P <0.05).Conclusion:Treg/Th17 immune imbalance exists in the mice with obese asthma.EGCG could inhibit the airway inflammation and hyperresponsiveness in the mice with obese asthma and improve the Treg/Th17 immune imbalance.
Objective:To investigate the regulatory effect of epigallocatechin-3-gallate(EGCG)on the immune imbalance of regulatory T lymphocytes/helper T lymphocytes 17(Treg/Th17)in the mice with obese asthma,and to provide the basis for the treatment of obese asthma by EGCG.Methods:A total of 40 C57BL/6J mice were randomly divided into normal control group(fed with chow diet),obese group(fed with high fat diet),obese asthma group [fed with high fat diet and sensitized and challenged with ovalbumin(ova)],EGCG intervention group(20 mg·kg~(-1) EGCG was administered intraperitoneally 1 hbefore challenge of OVA)and dexamethasone intervention group(2 mg·kg~(-1) dexamethasone was administered intraperitoneally 1 hbefore challenge of OVA),and there were 8 mice in each group.The edhanced pause(Penh)values of the mice in various groups were measured with non-invasive lung function instrument;the pathomorphology of lung tissue of the mice in various groups were observed by HE staining,and the inflammation scores were evaluated by semi-quantitative method for HE staining section;the levels of interleukin~(-1)0(IL~(-1)0)and interleukin~(-1)7 A(IL~(-1)7 A)in bronchoalveolar lavage fluid(BALF)and adiponectin and leptin in serum of the mice in various groups were detected by ELISA method,and the percentages of Treg and Th17 in spleen tissue of the mice in various groups were detected by flow cytometery.Results:Compared with normal control group,the body weight of the mice in obese group was increased(P<0.05);it was 1.67 times the average weight of the mice in normal control group.Compared with normal control group,the Penh value and inflammation score of the mice in obese asthma group were increased(P<0.05);the leptin level in serum of the mice in obese asthma group was increased(P<0.05);the IL~(-1)7 level in BALF of the mice in obese asthma group was increased(P<0.05)and IL~(-1)0 level in BALF of the mice in obese asthma group was decreased(P<0.05);the percentage of Th17 in spleen tissue of the mice in obese asthma group was increased(P<0.05)and the percentage of Treg in spleen tissue of the mice in obese asthma group was decreased(P<0.05).Compared with obese asthma group,the Penh value and inflammation score of the mice in EGCG intervention group were decreased(P<0.05);the leptin level in serum of the mice in EGCG intervention group was decreased(P<0.05);the IL~(-1)7 level in BALF of the mice in EGCG intervention group was decreased(P<0.05)and IL~(-1)0 level in BALF of the mice in EGCG intervention group was increased(P<0.05);the percentage of Th17 in spleen tissue of the mice in EGCG intervention group was decreased(P<0.05)and the percentage of Treg in spleen tissue of the mice in EGCG intervention group was increased(P <0.05).Conclusion:Treg/Th17 immune imbalance exists in the mice with obese asthma.EGCG could inhibit the airway inflammation and hyperresponsiveness in the mice with obese asthma and improve the Treg/Th17 immune imbalance.
引文
[1]MONIKO M,PAWLICZAK R.Obesity and asthma:risk,control and treatment[J].Postepy Dermatol Alergol,2018,35(6):563-571.
[2]Global strategy for asthma management and prevention.2014Global Initiative for Asthma[R/OL].http://www.ginasthma.Org/.2014.8.12.
[3]SASAKI M,YOSHIDA K,ADACHI Y,et al.Factors associated with asthma control in children:findings from a national Web-based survey[J].Pediatr Allergy Immunol,2014,25(8):804-809.
[4]ZHAO J,LLOYD C M,NOBLE A.Th17responses in chronic allergic airway inflammation abrogate regulatory T cellmediated tolerance and contribute to airway remodeling[J].Mucosal Immunol,2013,6(2):335-346.
[5]JIANG H,WU X B,ZHU H Y,et al.FOXP3(+)Treg/Th17cell imbalance in lung tissues of mice with asthma[J].Int J Clin Exp Med,2015,8(3):4158-4163.
[6]JOEL A M,ALLISON P W,LUIZA R,et al.Induction of IL-17Aprecedes development of airway hyperresponsiveness during diet-induced obesity and correlates with complement factor D[J].Front Immunol,2014,9(5):440.
[7]METIN D,ERKUT K,BURCU O,et al.CD4+,CD25+,FOXP3+T regulatory cell levels in obese,asthmatic,asthmatic obese,and healthy children[J].Inflammation,2015,38(4):1473-1478.
[8]GUPTA R K,GUPTA K,DWIVEDI P D.Pathophysiology of IL-33and IL-17in allergic disorders[J].Cytokine Growth Factor Rev,2017,38:22-36.
[9]ORDOEZ C L,SHAUGHNESSY T E,MATTHAY M A,et al.Increased neutrophil numbers and IL-8levels in airway secretions in acute severe asthma:Clinical and biologic significance[J].Am J Respir Crit Care Med,2000,161(4Pt 1):1185-1190.
[10]ITO K,HERBERT C,SIEGLE J S,et al.Steroid-resistant neutrophilic inflammation in a mouse model of an acute exacerbation of asthma[J].Am J Respir Cell Mol Biol,2008,39(5):543-550.
[11]YANG N,ZHANG H,CAI X X,et al.Epigallocatechin-3-gallate inhibits inflammation and epithelial-mesenchymal transition through the PI3K/AKT pathway via upregulation of PTEN in asthma[J].Int J Mol Med,2018,41(2):818-828.
[12]BYUN J K,YOON B Y,JHUNA J Y,et al.Epigallocatechin-3-gallate ameliorates both obesity and autoinflammatory arthritis aggravated by obesity by altering the balance among CD4+T-cell subsets[J].Immunol Lett,2014,157(1/2):51-59.
[13]HENDERSON W R JR,TANG L O,CHU S J,et al.A role for cysteinyl leukotrienes in airway remodeling in a mouse asthma model[J].Am J Respir Crit Care Med,2002,165(1):108-116.
[14]KIM H Y,LEE H J,CHANG Y J,et al.Interleukin-17-producing innate lymphoid cells and the NLRP3inflammasome facilitate obesity-associated airway hyperreactivity[J].Nat Med,2014,20(1):54-61.
[15]JUNG S H,KWON J M,SHIM J W,et al.Effects of dietinduced mild obesity on airway hyperreactivity and lung inflammation in mice[J].Yonsei Med J,2013,54(6):1430-1437.
[16]PARK H J,LEE J H,PARK Y H,et al.Roflumilast ameliorates airway hyperresponsiveness caused by diet-induced obesity in a murine model[J].Am J Respir Cell Mol Biol,2016,55(1):82-91.
[17]SNCHEZ-ZAUCO N,DEL RIO-NAVARRO B,GALLARDO-CASAS C,et al.High expression of Toll-like receptors 2 and 9 and Th1/Th2 cytokines profile in obese asthmatic children[J].Allergy Asthma Proc,2014,35(3):34-41.
[18]LEIRIA L O,MARTINS M A,SAAD M J.Obesity and asthma:beyond T(H)2inflammation[J].Metab Clin Exp,2015,64(2):172-181.
[19]SCOTT H A,GIBSON P G,GARG M L,et al.Airway inflammation is augmented by obesity and fatty acids in asthma[J].Eur Respir J,2011,38(3):594-602.
[20]TELENGA E D,TIDEMAN S W,KERSTJENS H A,et al.Obesity in asthma:more neutrophilic inflammation as a possible explanation for a reduced treatment response[J].Allergy,2012,67(8):1060-1068.
[21]MATHEWS J A,WURMBRAND A P,NETO F L,et al.Induction of IL-17A precedes development of airway hyperresponsiveness during diet induced obesity and correlates with complement factor D[J].Front Immunol,2014,15(5):440.
[22]KANELLOPOULOU C,MULJO S A.Fine-tuning Th17cells:to be or not to be pathogenic[J].Immunity,2016,44(6):1241-1243.
[23]ZHANG W,ZHANG X,SHENG A,et al.γ-Secretase gamma-secretase inhibitor alleviates acute airway inflammation of allergic asthma in mice by downregulating Th17 cell differentiation[J].Mediators Inflam,2015:258168.
[24]LISHEN D,XINYUAN K,FEN L,et al.Epigallocatechin gallate improves airway inflammation through TGF-β1signaling pathway in asthmatic mice[J].Mol Med Rep,2018,18(2):2088-2096.
[25]CHOI Y S,BAE C H,SONG S Y,et al.The effect of Epigallocatechin-3-gallate in allergic airway inflammation[J].Rhinology,2014,52(4):406-412.
[2]Global strategy for asthma management and prevention.2014Global Initiative for Asthma[R/OL].http://www.ginasthma.Org/.2014.8.12.
[3]SASAKI M,YOSHIDA K,ADACHI Y,et al.Factors associated with asthma control in children:findings from a national Web-based survey[J].Pediatr Allergy Immunol,2014,25(8):804-809.
[4]ZHAO J,LLOYD C M,NOBLE A.Th17responses in chronic allergic airway inflammation abrogate regulatory T cellmediated tolerance and contribute to airway remodeling[J].Mucosal Immunol,2013,6(2):335-346.
[5]JIANG H,WU X B,ZHU H Y,et al.FOXP3(+)Treg/Th17cell imbalance in lung tissues of mice with asthma[J].Int J Clin Exp Med,2015,8(3):4158-4163.
[6]JOEL A M,ALLISON P W,LUIZA R,et al.Induction of IL-17Aprecedes development of airway hyperresponsiveness during diet-induced obesity and correlates with complement factor D[J].Front Immunol,2014,9(5):440.
[7]METIN D,ERKUT K,BURCU O,et al.CD4+,CD25+,FOXP3+T regulatory cell levels in obese,asthmatic,asthmatic obese,and healthy children[J].Inflammation,2015,38(4):1473-1478.
[8]GUPTA R K,GUPTA K,DWIVEDI P D.Pathophysiology of IL-33and IL-17in allergic disorders[J].Cytokine Growth Factor Rev,2017,38:22-36.
[9]ORDOEZ C L,SHAUGHNESSY T E,MATTHAY M A,et al.Increased neutrophil numbers and IL-8levels in airway secretions in acute severe asthma:Clinical and biologic significance[J].Am J Respir Crit Care Med,2000,161(4Pt 1):1185-1190.
[10]ITO K,HERBERT C,SIEGLE J S,et al.Steroid-resistant neutrophilic inflammation in a mouse model of an acute exacerbation of asthma[J].Am J Respir Cell Mol Biol,2008,39(5):543-550.
[11]YANG N,ZHANG H,CAI X X,et al.Epigallocatechin-3-gallate inhibits inflammation and epithelial-mesenchymal transition through the PI3K/AKT pathway via upregulation of PTEN in asthma[J].Int J Mol Med,2018,41(2):818-828.
[12]BYUN J K,YOON B Y,JHUNA J Y,et al.Epigallocatechin-3-gallate ameliorates both obesity and autoinflammatory arthritis aggravated by obesity by altering the balance among CD4+T-cell subsets[J].Immunol Lett,2014,157(1/2):51-59.
[13]HENDERSON W R JR,TANG L O,CHU S J,et al.A role for cysteinyl leukotrienes in airway remodeling in a mouse asthma model[J].Am J Respir Crit Care Med,2002,165(1):108-116.
[14]KIM H Y,LEE H J,CHANG Y J,et al.Interleukin-17-producing innate lymphoid cells and the NLRP3inflammasome facilitate obesity-associated airway hyperreactivity[J].Nat Med,2014,20(1):54-61.
[15]JUNG S H,KWON J M,SHIM J W,et al.Effects of dietinduced mild obesity on airway hyperreactivity and lung inflammation in mice[J].Yonsei Med J,2013,54(6):1430-1437.
[16]PARK H J,LEE J H,PARK Y H,et al.Roflumilast ameliorates airway hyperresponsiveness caused by diet-induced obesity in a murine model[J].Am J Respir Cell Mol Biol,2016,55(1):82-91.
[17]SNCHEZ-ZAUCO N,DEL RIO-NAVARRO B,GALLARDO-CASAS C,et al.High expression of Toll-like receptors 2 and 9 and Th1/Th2 cytokines profile in obese asthmatic children[J].Allergy Asthma Proc,2014,35(3):34-41.
[18]LEIRIA L O,MARTINS M A,SAAD M J.Obesity and asthma:beyond T(H)2inflammation[J].Metab Clin Exp,2015,64(2):172-181.
[19]SCOTT H A,GIBSON P G,GARG M L,et al.Airway inflammation is augmented by obesity and fatty acids in asthma[J].Eur Respir J,2011,38(3):594-602.
[20]TELENGA E D,TIDEMAN S W,KERSTJENS H A,et al.Obesity in asthma:more neutrophilic inflammation as a possible explanation for a reduced treatment response[J].Allergy,2012,67(8):1060-1068.
[21]MATHEWS J A,WURMBRAND A P,NETO F L,et al.Induction of IL-17A precedes development of airway hyperresponsiveness during diet induced obesity and correlates with complement factor D[J].Front Immunol,2014,15(5):440.
[22]KANELLOPOULOU C,MULJO S A.Fine-tuning Th17cells:to be or not to be pathogenic[J].Immunity,2016,44(6):1241-1243.
[23]ZHANG W,ZHANG X,SHENG A,et al.γ-Secretase gamma-secretase inhibitor alleviates acute airway inflammation of allergic asthma in mice by downregulating Th17 cell differentiation[J].Mediators Inflam,2015:258168.
[24]LISHEN D,XINYUAN K,FEN L,et al.Epigallocatechin gallate improves airway inflammation through TGF-β1signaling pathway in asthmatic mice[J].Mol Med Rep,2018,18(2):2088-2096.
[25]CHOI Y S,BAE C H,SONG S Y,et al.The effect of Epigallocatechin-3-gallate in allergic airway inflammation[J].Rhinology,2014,52(4):406-412.