Qingfei Xiaoyan Wan alleviates asthma through multi-target network regulation

BMC Complementary and Alternative Medicine

Hong Wu¹  Gang Bai³  Binfeng Cheng³  Pengwei Pan³  Yingbo Miao³  Zhenying Zhao² 

[1] Department of Pharmacology, Tianjin Medical University, Tianjin, People's Republic of China;Department of Pharmacy, Tianjin People’s Hospital, Tianjin, People's Republic of China;College of Pharmacy, Nankai University, Tianjin, People's Republic of China

关键词: Multi-target network;    MS identification;    2D- electrophoresis;    Microarray;    Traditional Chinese Medicine;    Asthma;   

Others  :  1221036
DOI  :  10.1186/1472-6882-13-206

 received in 2013-03-25, accepted in 2013-08-02,  发布年份 2013

【 摘 要 】

Background

Qingfei Xiaoyan Wan (QFXY), a traditional Chinese formula, is widely used for relieving cough, asthma, upper respiratory tract infection, bronchitis, pneumonia, and etc. in clinic. Comparing with other anti-asthma drugs, it is characterised with moderate and persistent efficacy as well as few side effects, however, the underlying action mechanism still remains elusive. This study aimed to identify QFXY multi-target network regulation as an asthma controller.

Methods

This study established asthma model induced by histamine phosphate and acetylcholine chloride (His&Ach) in guinea pigs, which then were administered orally with QFXY. Hematoxylin-Eosin staining sections were applied for evaluating QFXY effect. In both Model and QFXY groups, customized microarrays and 2D electrophoresis were adopted to detect differentially expressed genes (diff genes) and proteins (diff proteins) respectively, and some diff proteins were identified with MALDI-TOF/MS. The checked diff genes and proteins underwent Cluster, GO and KEGG analysis. Based on GAD and HPRD databases, QFXY-asthma target regulation network was constructed.

Results

His&Ach-induced asthma model of guinea pigs was established. HE sections presented anti-inflammation and anti-remodelling effects of QFXY. Comparing with the Model group, 55 diff genes and 6 diff proteins were identified in QFXY group. Validation by qPCR and Western blot showed the microarray and 2D data reliable. Furthermore, QFXY-asthma target regulation network was achieved.

Conclusions

A primarily combined genomic and proteomic screening of QFXY targets displayed a series of candidate genes and proteins, which indicated that the effect of QFXY relied on the combined mechanism, anti-inflammation and anti-remodelling, as well as influencing signal transduction in vivo.

【 授权许可】

   
2013 Zhao et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Bateman ED, Hurd SS, Barnes PJ, Bousquet J, Drazen JM, FitzGerald M, Gibson P, Ohta K, O'Byrne P, Pedersen SE, Pizzichini E, Sullivan SD, Wenzel SE, Zar HJ: Global strategy for asthma management and prevention: GINA executive summary. Eur Respir J 2008, 31(1):143-178.
• [2]Holgate ST, Davies DE: Rethinking the pathogenesis of asthma. Immunity 2009, 31(3):362-367.
• [3]Gern JE, Lemanske RF, Busse WW: Early life origins of asthma. J Clin Invest 1999, 104(7):837-843.
• [4]Claudio L, Tulton L, Doucette J, Landrigan PJ: Socioeconomic factors and asthma hospitalization rates in New York City. J Asthma 1999, 36(4):327-343.
• [5]Bousquet J, Jeffery PK, Busse WW, Johnson M, Vignola AM: Asthma From bronchoconstriction to airways inflammation and remodeling. Am J Respir Crit Care Med 2000, 161(5):1720-1745.
• [6]Kroegel C: Global Initiative for Asthma (GINA) guidelines: 15 years of application. Expert Rev Clin Immunol 2009, 5(3):239-249.
• [7]Dorinsky : The salmeterol multicenter asthma research trial: A comparison of usual pharmacotherapy for asthma or usual pharmacotherapy plus salmeterol (vol 129, pg 15, 2006). Chest 2006, 129(5):1393.
• [8]Gibson PG, Powell H, Ducharme FM: Differential effects of maintenance long-acting beta-agonist and inhaled corticosteroid on asthma control and asthma exacerbations. J Allergy Clin Immun 2007, 119(2):344-350.
• [9]Lemanske RF, Sorkness CA, Mauger EA, Lazarus SC, Boushey HA, Fahy JV, Drazen JM, Chinchilli VM, Craig T, Fish JE, Ford JG, Israel E, Kraft M, Martin RJ, Nachman SA, Peters SP, Spahn JD, Szefler SJ: Inhaled corticosteroid reduction and elimination in patients with persistent asthma receiving salmeterol - A randomized controlled trial. Jama-J Am Med Assoc 2001, 285(20):2594-2603.
• [10]Wechsler ME, Pauwels R, Drazen JM: Leukotriene modifiers and Churg-Strauss syndrome - Adverse effect or response to corticosteroid withdrawal? Drug Saf 1999, 21(4):241-251.
• [11]Barnes PJ: Theophylline - New perspectives for an old drug. Am J Resp Crit Care 2003, 167(6):813-818.
• [12]Hou Y, Li R, Shi Q, Cheng B, Wang L, Bai R: Investigation of the Anti-asthmatic Mechanism of QingFeiXiaoYanWan. Acta Scientiarum Naturalium Universitatis Nankaiensis 2009, 5:103-107.
• [13]Cheng B, Hou Y, Wang L, Dong L, Peng J, Bai G: Dual-bioactivity-based liquid chromatography-coupled quadrupole time-of-flight mass spectrometry for NF-kappaB inhibitors and beta2AR agonists identification in Chinese Medicinal Preparation Qingfei Xiaoyan Wan. Anal Bioanal Chem 2012, 404(8):2445-2452.
• [14]Vogel G, Vogel WH, Vogel HG: Drug Discovery and Evaluation: Pharmacological Assays. Berlin: Springer; 1997.
• [15]Fan X, Shi M, Wang Y, Liang Q, Luo G: Transcriptional profiling analysis of HMP-treated rats with experimentally induced myocardial infarction. J Ethnopharmacol 2011, 137(1):199-204.
• [16]Mutter GL, Zahrieh D, Liu CM, Neuberg D, Finkelstein D, Baker HE, Warrington JA: Comparison of frozen and RNALater solid tissue storage methods for use in RNA expression microarrays. BMC Genomics 2004, 5:88-94. BioMed Central Full Text
• [17]Pedotti P, Hoen PA T, Vreugdenhil E, Schenk GJ, Vossen RH, Ariyurek Y, De Hollander M, Kuiper R, Van Ommen G, Den Dunnen JT, Boer JM, De Menezes RX: Can subtle changes in gene expression be consistently detected with different microarray platforms? BMC Genomics 2008, 9:124-136. BioMed Central Full Text
• [18]Basic BLAST. http://blast.ncbi.nlm.nih.gov/Blast.cgi webcite
• [19]Molecule Annotation System. http://bioinfo.capitalbio.com/mas3/ webcite
• [20]Fan X, Li X, Lv S, Wang Y, Zhao Y, Luo G: Comparative proteomics research on rat MSCs differentiation induced by Shuanglong Formula. J Ethnopharmacol 2010, 131(3):575-580.
• [21]Human Protein Reference Database. http://www.hprd.org/ webcite
• [22]Genetic Association Database. http://geneticassociationdb.nih.gov/ webcite
• [23]Cline MS, Smoot M, Cerami E, Kuchinsky A, Landys N, Workman C, Christmas R, Avila-Campilo I, Creech M, Gross B, Hanspers K, Isserlin R, Kelley R, Killcoyne S, Lotia S, Maere S, Morris J, Ono K, Pavlovic V, Pico AR, Vailaya A, Wang PL, Adler A, Conklin BR, Hood L, Kuiper M, Sander C, Schmulevich I, Schwikowski B, Warner GJ, et al.: Integration of biological networks and gene expression data using Cytoscape. Nat Protoc 2007, 2(10):2366-2382.
• [24]NCBI EST database. http://www.ncbi.nlm.nih.gov/nucest/ webcite
• [25]KEGG PATHWAY Database. http://www.genome.jp/kegg/pathway.html webcite
• [26]GenMapp: Gene Map Annotation and Pathway Profiler. http://www.genmapp.org/default.html webcite
• [27]BioCarta. http://www.biocarta.com/genes/index.asp webcite
• [28]Bates JH, Rincon M, Irvin CG: Animal models of asthma. Am J Physiol Lung Cell Mol Physiol 2009, 297(3):L401-L410.
• [29]Canning BJ: Modeling asthma and COPD in animals: a pointless exercise? Curr Opin Pharmacol 2003, 3(3):244-250.
• [30]Szelenyi I: Animal models of bronchial asthma. Inflamm Res 2000, 49(12):639-654.
• [31]Holgate ST: Pathogenesis of asthma. Clin Exp Allergy 2008, 38(6):872-897.
• [32]Kitanaka J, Kitanaka N, Takemura M, Wang XB, Hembree CM, Goodman NL, Uhl GR: Isolation and Sequencing of a putative promoter region of the murine G protein beta 1 subunit (GNB1) gene. Dna Sequence 2002, 13(1):39-45.
• [33]Moya-Quiles MR, Ballesta-Martinez MJ, Lopez-Gonzalez V, Glover G, Guillen-Navarro E: A compound heterozygous mutation in the EDAR gene in a Spanish family with autosomal recessive hypohidrotic ectodermal dysplasia. Arch Dermatol Res 2010, 302(4):307-310.
• [34]Shaul YD, Seger R: The MEK/ERK cascade: From signaling specificity to diverse functions. Bba-Mol Cell Res 2007, 1773(8):1213-1226.
• [35]McCubrey JA, Milella M, Tafuri A, Martelli AM, Lunghi P, Bonati A, Cervello M, Lee JT, Steelman LS: Targeting the Raf/MEK/ERK pathway with small-molecule inhibitors. Curr Opin Investig Drugs 2008, 9(6):614-630.
• [36]Verhaeghe C, Remouchamps C, Hennuy B, Vanderplasschen A, Chariot A, Tabruyn SP, Oury C, Bours V: Role of IKK and ERK pathways in intrinsic inflammation of cystic fibrosis airways. Biochem Pharmacol 2007, 73(12):1982-1994.
• [37]Perkins ND: Post-translational modifications regulating the activity and function of the nuclear factor kappa B pathway. Oncogene 2006, 25(51):6717-6730.
• [38]Weiss T, Shalit I, Blau H, Werber S, Halperin D, Levitov A, Fabian I: Anti-inflammatory effects of moxifloxacin on activated human monocytic cells: Inhibition of NF-KB and mitogen-activated protein kinase activation and of synthesis of proinflammatory ctokines. Antimicrob Agents Ch 2004, 48(6):1974-1982.
• [39]Lee FS, Peters RT, Dang LC, Maniatis T: MEKK1 activates both I kappa B kinase alpha and I kappa B kinase beta. P Natl Acad Sci Usa 1998, 95(16):9319-9324.
• [40]Fontana J, Fulton D, Chen Y, Fairchild TA, McCabe TJ, Fujita N, Tsuruo T, Sessa WC: Domain mapping studies reveal that the M domain of hsp90 serves as a molecular scaffold to regulate Akt-dependent phosphorylation of endothelial nitric oxide synthase and NO release. Circ Res 2002, 90(8):866-873.
• [41]Ozes ON, Mayo LD, Gustin JA, Pfeffer SR, Pfeffer LM, Donner DB: NF-kappa B activation by tumour necrosis factor requires the Akt serine-threonine kinase. Nature 1999, 401(6748):82-85.
• [42]Sato S, Fujita N, Tsuruo T: Modulation of Akt kinase activity by binding to Hsp90. P Natl Acad Sci Usa 2000, 97(20):10832-10837.
• [43]Gettins P, Olson ST: Exosite Determinants of Serpin Specificity. J Biol Chem 2009, 284(31):20441-20445.
• [44]Ekeowa UI, Marciniak SJ, Lomas DA: alpha(1)-antitrypsin deficiency and inflammation. Expert Rev Clin Immunol 2011, 7(2):243-252.
• [45]Janciauskiene SM, Bals R, Koczulla R, Vogelmeier C, Kohnlein T, Welte T: The discovery of alpha 1-antitrypsin and its rote in health and disease. Resp Med 2011, 105(8):1129-1139.
• [46]Shigetomi H, Onogi A, Kajiwara H, Yoshida S, Furukawa N, Haruta S, Tanase Y, Kanayama S, Noguchi T, Yamada Y, Oi H, Kobayashi H: Anti-inflammatory actions of serine protease inhibitors containing the Kunitz domain. Inflamm Res 2010, 59(9):679-687.
• [47]Zhang B, Hu Y, Ma JX: Anti-inflammatory and antioxidant effects of SERPINA3K in the retina. Invest Ophthalmol Vis Sci 2009, 50(8):3943-3952.
• [48]Meyer N, Akdis CA: Vascular Endothelial Growth Factor as a Key Inducer of Angiogenesis in the Asthmatic Airways. Curr Allergy Asthma Rep 2013, 13(1):1-9.
• [49]Budd DC, Holmes AM: Targeting TGF beta superfamily ligand accessory proteins as novel therapeutics for chronic lung disorders. Pharmacol Therapeut 2012, 135(3):279-291.
• [50]Engelse MA, Neele JA, Bronckers A, Pannekoek H, De Vries C: Vascular calcification: expression patterns of the osteoblast-specific gene core binding factor alpha-1 and the protective factor matrix gla protein in human atherogenesis. Cardiovasc Res 2001, 52(2):281-289.
• [51]Bostrom K, Zebboudj AF, Yao YC, Lin TS, Torres A: Matrix GLA protein stimulates VEGF expression through increased transforming growth factor-beta 1 activity in endothelial cells. J Biol Chem 2004, 279(51):52904-52913.
• [52]Boulet LP, Turcotte H, Laviolette M, Naud F, Bernier MC, Martel S, Chakir J: Airway hyperresponsiveness, inflammation, and subepithelial collagen deposition in recently diagnosed versus long-standing mild asthma - Influence of inhaled corticosteroids. Am J Resp Crit Care 2000, 162(4):1308-1313.
• [53]Vignola AM, Mirabella F, Costanzo G, Di Giorgi R, Gjomarkaj M, Bellia V, Bonsignore G: Airway remodeling in asthma. Chest 2003, 123S(3):417S-422S.
• [54]Burgess JK: The role of the extracellular matrix and specific growth factors in the regulation of inflammation and remodelling in asthma. Pharmacol Therapeut 2009, 122(1):19-29.