铁线蕨总黄酮对哮喘大鼠模型及其尿液代谢物的影响
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摘要
目的探讨铁线蕨总黄酮(TXJF)对哮喘模型大鼠及其尿液代谢物的影响。方法选取60只健康雄性SD大鼠,采用OVA致敏及雾化吸入方法,建立哮喘大鼠模型,随机分为正常对照组,阳性对照组(醋酸地塞米松),哮喘模型组,铁线蕨总黄酮(TXJF)组(高、中、低剂量)各组10只,TXJF组以TXJF进行灌胃给药,检测各组大鼠肺泡灌洗液(BALF)中的炎症细胞数目及分类,肺组织病理形态学改变,测定血清免疫球蛋白E(IgE),从而进行模型的评价及观察TXJF对哮喘模型大鼠的影响。收集大鼠尿液,利用核磁共振波普(1 H-NMR)技术检测氢谱,从而建立大鼠尿液代谢指纹谱,正交偏最小二乘判别分析法(OPLS-DA),运用偏最小二乘判别分析法(PLS-DA)观察各组大鼠尿液中代谢产物的差异性。结果与正常对照组相比较,哮喘模型组大鼠出现哮喘样症状,BALF中细胞总数及炎症细胞百分率升高,肺组织出现病理学改变,血清IgE水平升高;TXJF组大鼠哮喘样症状有所好转,BALF中细胞总数及炎症细胞百分率下降、肺组织病理改变情况减轻;与哮喘模型组比较,TXJF组血清IgE水平下降。哮喘模型组尿液中异亮氨酸、柠檬酸、琥珀酸、甘氨酸、丙氨酸含量降低,乳酸含量升高。与哮喘模型组相比较,高剂量TXJF组中异亮氨酸、琥珀酸、柠檬酸、肌酸、牛磺酸、丙氨酸、甘氨酸含量升高,乳酸等含量降低,差异具有统计学意义(P<0.05)。结论 TXJF能够改善哮喘大鼠的一般情况,减轻肺组织病理改变,降低血清IgE水平,降低肺泡灌洗液(BALF)中的炎症细胞数目及分类,并且对哮喘大鼠尿液代谢物具有显著的影响,从而起到治疗哮喘的作用,其作用机制可能与作用于哮喘大鼠体内的氨基酸代谢、能量代谢等多种代谢途径有关联。
Objective To investigate the effect of total flavonoids of Adiantum capillus-veneris L.on asthma rat models and their urine metabolites.Methods Fifty healthy male Sprague-Dawley rats were to sensitize and inhale by OVA to establish an asthmatic rat model.The rats were randomly divided into normal control group,positive control group(dexamethasone acetate group),asthma model group,and TXJF(high,medium,and low-dose)groups(each group has 10 rats),and TXJF was administered intragastrically.The number and classification of inflammatory cells in bronchoalveolar lavage fluid(BALF),the histopathological changes of lung tissue and the serum IgE level were detected to model evaluation and observe the effect of TXJF on asthma model rats.The rat urine and the proton spectrum was detected by nuclear magnetic resonance(1 H-NMR)technique to establish the metabolic fingerprint of rat urine.The difference in metabolites between urine metabolites of rats in each group were observed using the Orthogonal Partial Least Squares Discriminant Analysis(OPLS-DA)and Partial Least Squares Discriminant Analysis(PLS-DA).Results Compared with the normal control group,the asthma model group showed asthma-like symptoms,the total number of cells and inflammatory cells in the BALF increased,the pathological changes appeared in the lung tissue,serum IgE levels increased;asthma-like symptoms in the TXJF group rats were improved,the percentage of total cells and inflammatory cells in the BALF decreased,and the pathological changes in the lung tissue alleviated.Compared with the asthma model group,serum IgE levels in the TXJF group decreased.The levels of isoleucine,citric acid,succinic acid,glycine,and alanine in the urine of the asthma model group decreased,and the lactate content increased.Compared with the asthma model group,the content of isoleucine,succinic acid,citric acid,creatine,taurine,alanine,and glycine in the high-dose TXJF group increased,and the content of lactic acid decreased.The difference was statistically significant(P <0.05).Conclusion TXJF can improve the general condition of asthmatic rats,reduce the pathological changes of lung tissue,reduce the level of serum IgE,decrease the number and classification of inflammatory cells in BALF,and have significant effect on urinary metabolites in asthmatic rats,thus it plays a role in the treatment of asthma.The mechanism of action may be related to various metabolic pathways such as amino acid metabolism and energy metabolism in asthmatic rats.
Objective To investigate the effect of total flavonoids of Adiantum capillus-veneris L.on asthma rat models and their urine metabolites.Methods Fifty healthy male Sprague-Dawley rats were to sensitize and inhale by OVA to establish an asthmatic rat model.The rats were randomly divided into normal control group,positive control group(dexamethasone acetate group),asthma model group,and TXJF(high,medium,and low-dose)groups(each group has 10 rats),and TXJF was administered intragastrically.The number and classification of inflammatory cells in bronchoalveolar lavage fluid(BALF),the histopathological changes of lung tissue and the serum IgE level were detected to model evaluation and observe the effect of TXJF on asthma model rats.The rat urine and the proton spectrum was detected by nuclear magnetic resonance(1 H-NMR)technique to establish the metabolic fingerprint of rat urine.The difference in metabolites between urine metabolites of rats in each group were observed using the Orthogonal Partial Least Squares Discriminant Analysis(OPLS-DA)and Partial Least Squares Discriminant Analysis(PLS-DA).Results Compared with the normal control group,the asthma model group showed asthma-like symptoms,the total number of cells and inflammatory cells in the BALF increased,the pathological changes appeared in the lung tissue,serum IgE levels increased;asthma-like symptoms in the TXJF group rats were improved,the percentage of total cells and inflammatory cells in the BALF decreased,and the pathological changes in the lung tissue alleviated.Compared with the asthma model group,serum IgE levels in the TXJF group decreased.The levels of isoleucine,citric acid,succinic acid,glycine,and alanine in the urine of the asthma model group decreased,and the lactate content increased.Compared with the asthma model group,the content of isoleucine,succinic acid,citric acid,creatine,taurine,alanine,and glycine in the high-dose TXJF group increased,and the content of lactic acid decreased.The difference was statistically significant(P <0.05).Conclusion TXJF can improve the general condition of asthmatic rats,reduce the pathological changes of lung tissue,reduce the level of serum IgE,decrease the number and classification of inflammatory cells in BALF,and have significant effect on urinary metabolites in asthmatic rats,thus it plays a role in the treatment of asthma.The mechanism of action may be related to various metabolic pathways such as amino acid metabolism and energy metabolism in asthmatic rats.
引文
[1]吴彩霞,张勇,顾雪竹,等.铁线蕨化学成分研究和药理作用研究进展[J].中国医药导报,2014,11(2):39-40.
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[7]KASABRI V,ALHALLAQ E K,BUSTANJI Y K,et al.Antiobesity and antihyperglycaemic effects of Adiantum capillusveneris extracts:in vitro and in vivo evaluations[J].Pharm Biol,2017,55(1):164-172.
[8]阿迪拉木·阿依甫江,麦合苏木·艾克木,古孜力努尔·依马木,等.维药铁线蕨总黄酮抗炎平喘作用研究[J].新疆医科大学学报,2016,39(1):17-20.
[9]陈馨馨,李友林.基于肺脏特性中药干预哮喘缓解期家兔的尿液代谢组学研究[J].中国老年保健医学,2017,15(3):5-7.
[10]TAHERI S,ALIANNEJED R,REZAEI-TAVIRANI M,et al.The metabolomics of airway diseases,including COPD,asthma and cystic fibrosis[J].Biomarkers,2015,20(1):15-16.
[11]SAUDE E J,OBEFUNA I P,SOMORJAI R L,et al.Metabolomic biomarkers in a model of asthma exacerbation:urine nuclear magnetic resonance[J].Am J Respir Crit Care Med,2009,179(1):25-34.
[12]阿迪拉木·阿依甫江.维药铁线蕨总黄酮的提取、纯化及药效学研究[D].乌鲁木齐:新疆医科大学,2016.
[13]LINDBERG W,PERSSON J,WOLD S,et al.Partial leastsquares method for spectrofluorimetric analysis of mixtures of humic acid and lignin sulfonate[J].Analy Chem,1983,55(4):643-648.
[14]TRYGG J,WOLD S.Orthogonal projections to latent structures(OPLS)[J].J Chemometrics,2002,16(3):119-128.
[2]中华人民共和国卫生部药典委员会.中华人民共和国卫生部药品标准[S].乌鲁木齐:新疆科技卫生出版社,1999:312-313.
[3]SANTHOSH K S,NAGARAJAN N.Screening of preliminary phytochemical constituents and antimicrobial activity of Adiantum capillus-veneris L.[J].Res Antimicmb,2012,16(1):56-61.
[4]GUHA P,MUKHOPADHYAY R,GUPTA K,et al.Antifungal activity of the crude extracts and extracted phenols from gametophytes and sporophytes of two species of Adiantum[J].Taiwania,2005,50(4):272-283.
[5]IBRAHEIM Z Z,AHMED A S,GOUDA Y G,et al.Phytochemical and biological studies of Adiantum capillus-veneris L.[J].Saudi Pharm J,2011,19(2):65-74.
[6]SINGH M,SINGH N,KHARE P B,et al.Antimicrobial activity of some important Adiantum species used traditionally in indigenous systems of medicine[J].J Ethno Pharm,2008,115(2):327-329.
[7]KASABRI V,ALHALLAQ E K,BUSTANJI Y K,et al.Antiobesity and antihyperglycaemic effects of Adiantum capillusveneris extracts:in vitro and in vivo evaluations[J].Pharm Biol,2017,55(1):164-172.
[8]阿迪拉木·阿依甫江,麦合苏木·艾克木,古孜力努尔·依马木,等.维药铁线蕨总黄酮抗炎平喘作用研究[J].新疆医科大学学报,2016,39(1):17-20.
[9]陈馨馨,李友林.基于肺脏特性中药干预哮喘缓解期家兔的尿液代谢组学研究[J].中国老年保健医学,2017,15(3):5-7.
[10]TAHERI S,ALIANNEJED R,REZAEI-TAVIRANI M,et al.The metabolomics of airway diseases,including COPD,asthma and cystic fibrosis[J].Biomarkers,2015,20(1):15-16.
[11]SAUDE E J,OBEFUNA I P,SOMORJAI R L,et al.Metabolomic biomarkers in a model of asthma exacerbation:urine nuclear magnetic resonance[J].Am J Respir Crit Care Med,2009,179(1):25-34.
[12]阿迪拉木·阿依甫江.维药铁线蕨总黄酮的提取、纯化及药效学研究[D].乌鲁木齐:新疆医科大学,2016.
[13]LINDBERG W,PERSSON J,WOLD S,et al.Partial leastsquares method for spectrofluorimetric analysis of mixtures of humic acid and lignin sulfonate[J].Analy Chem,1983,55(4):643-648.
[14]TRYGG J,WOLD S.Orthogonal projections to latent structures(OPLS)[J].J Chemometrics,2002,16(3):119-128.