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Journal of Chinese Integrative Medicine ›› 2013, Vol. 11 ›› Issue (3): 195-205.doi: 10.3736/jintegrmed2013029

• Research Article • Previous Articles     Next Articles

Constituents of the anti-asthma herbal formula ASHMITM synergistically inhibit IL-4 and IL-5 secretion by murine Th2 memory cells, and eotaxin by human lung fibroblasts in vitro

Bolleddula Jayaprakasama,Nan Yanga, Ming-Chun Wenb, Rong Wangc, Joseph Goldfarbd, Hugh Sampsona, Xiu-Min Lia()   

  1. a. Center for Chinese Herbal Therapy for Asthma and Allergy, Department of Pediatrics, Icahn School ofMedicine at Mount Sinai, New York, NY 10029, USA
    b. Department of Pneumology, Weifang Asthma Hospital, Weifang 261041, Shandong Province, China
    c. Department of Human Genetics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA
    d. Department of Pharmacology and Systems Therapeutics, Icahn School of Medicine at Mount Sinai, NewYork, NY 10029, USA
  • Received:2013-01-22 Accepted:2013-04-07 Online:2013-05-10 Published:2013-05-15


Anti-asthma herbal medicine intervention (ASHMITM), a combination of three traditional Chinese medicinal herbs developed in our laboratory, has demonstrated efficacy in both mouse models of allergic asthma, and a double-blind placebo-controlled clinical trial in patients with asthma. This study was designed to determine if the anti-inflammatory effects of individual herbal constituents of ASHMITM exhibited synergy.


Effects of ASHMI and its components aqueous extracts of Lingzhi (Ganoderma lucidum), Kushen (Sophora flavescens) and Gancao (Glycyrrhiza uralensis), on Th2 cytokine secretion by murine memory Th2 cells (D10.G4.1) and eotaxin-1 secretion by human lung fibroblast (HLF-1) cells were determined by measuring levels in culture supernatants by enzyme-linked immunosorbent assay. Potential synergistic effects were determined by computing interaction indices from concentration-effect curve parameters.


Individual Lingzhi, Kushen and Gancao extracts and ASHMI (the combination of individual extracts) inhibited production of interleukin (IL)-4 and IL-5 by murine memory Th2 cells and eotaxin-1 production by HLF-1 cells. The mean 25%-inhibitory-concentration (IC25) values (mg/mL) for ASHMI, Lingzhi, Kushen and Gancao for IL-4 production were 30.9, 79.4, 123, and 64.6, respectively; for IL-5 production were 30.2, 263, 123.2 and 100, respectively; for eotaxin-1 were 13.2, 16.2, 30.2, and 25.1, respectively. The IC50 values (mg/mL) for ASHMI, Lingzhi, Kushen and Gancao for IL-4 production were 158.5, 239.9, 446.7, and 281.8, respectively; for eotaxin-1 were 38.1, 33.1, 100, and 158.5, respectively. The interaction indices of ASHMI constituents at IC25 were 0.35 for IL-4, 0.21 for IL-5 and 0.59 for eotaxin-1. The interaction indices at IC50 values were 0.50 for IL-4 and 0.62 for eotaxin-1 inhibition. Inhibition of IL-5 did not reach IC50 values. All interaction indices were below 1 which indicated synergy.


By comparing the interaction index values, we find that constituents in ASHMITM synergistically inhibited eotaxin-1 production as well as Th2 cytokine production.

Key words: Traditional Chinese medicine, Herbal medicine, Plant extracts, Anti-asthma herbal medicine intervention (ASHMI), Anti-asthmatic agents, Chemokine CCL11, Interleukin-4, Interleukin-5, in vitro


Figure 1

Consistency of HPLC fingerprints and major chemical constituents in different preparations of ASHMIA: HPLC fingerprints generated from standard ASHMITM, experimental ASHMI, and individual herb extracts. B: Characterization of chemical constituents of major corresponding peaks between standard ASHMITM, experimental ASHMI and individual herb extracts. Peaks 16 and 32 correspond to liquiritin and glycyrrhizin from Gancao; peaks 7 and 19 correspond to matrine and kushenol O from Kushen; peaks 26 and 27 correspond to ganoderic acid D and ganoderic acid A from Lingzhi. HPLC: High pressure liquid chromatography."

Figure 2

Effects of A) ASHMI; B) Lingzhi; C) Kushen; and D) Gancao on conalbumin-induced interleukin-4D10 cells were cultured for 72 h with or without herbal extracts in the presence of antigen conalbumin and antigen-presenting cells. Interleukin-4 level was measured from the supernatant by enzyme-linked immunosorbent assay and the percentage of inhibition was calculated with respect to untreated cells. Three replications were run at each concentration. Concentration-response curves were constructed using Prism 4.0 (GraphPad Software, San Diego, CA, USA). The dotted lines indicate the 95% confidence intervals."


Figure 3

Effects of A) ASHMI; B) Lingzhi; C) Kushen; and D) Gancao on conalbumin-induced interleukin-5D10 cells were cultured for 72 h with or without herbal extracts in the presence of antigen conalbumin-presenting cells. Interleukin-5 level was measured from the supernatant by enzyme-linked immunosorbent assay and percent inhibition was calculated with respect to negative control. Three replicates were run at each concentration. Concentration-response curves were constructed using Prism 4.0 (GraphPad Software, San Diego, CA, USA). The dotted lines indicate the 95% confidence intervals."


Figure 4

Effects of A) ASHMI; B) Lingzhi; C) Kushen; and D) Gancao on eotaxin-1 production by human lung fibroblastsHuman lung fibroblasts were cultured with or without herbal extracts for 48 h. Eotaxin-1 level in the culture supernatants was measured by enzyme-linked immunosorbent assay. The percentage of inhibition was calculated with respect to untreated cells. Three replicates were run at each concentration. Concentration-response curves were constructed using Prism 4.0 (GraphPad Software, San Diego, CA, USA). The dotted lines indicate the 95% confidence intervals."


[1] Centers for Disease Control and Prevention. National Center for Health Statistics: Asthma 2011.[ 2012-01-18]. .
[2] Eid N, Morton R, Olds B, Clark P, Sheikh S, Looney S . Decreased morning serum cortisol levels in children with asthma treated with inhaled fluticasone propionate[J]. Pediatrics, 2002,109(2):217-221
doi: 10.1542/peds.109.2.217
[3] Visser MJ, van der Veer E, Postma DS, Arends LR, de Vries TW, Brand PL, Duiverman EJ . Side-effects of fluticasone in asthmatic children: no effects after dose reduction[J]. Eur Respir J, 2004,24(3):420-425
doi: 10.1183/09031936.04.00023904
[4] Wen MC, Wei CH, Hu ZQ, Srivastava K, Ko J, Xi ST, Mu DZ, Du JB, Li GH, Wallenstein S, Sampson H, Kattan M, Li XM . Efficacy and tolerability of anti-asthma herbal medicine intervention in adult patients with moderate-severe allergic asthma[J]. J Allergy Clin Immunol, 2005,116(3):517-524
doi: 10.1016/j.jaci.2005.05.029
[5] Li XM . Complementary and alternative medicine in pediatric allergic disorders[J]. Curr Opin Allergy Clin Immunol, 2009,9(2):161-167
doi: 10.1097/ACI.0b013e328329226f
[6] Hsu CH, Lu CM, Chang TT . Efficacy and safety of modified Mai-Men-Dong-Tang for treatment of allergic asthma[J]. Pediatr Allergy Immunol, 2005,16(1):76-81
doi: 10.1111/pai.2005.16.issue-1
[7] Chan CK, Kuo ML, Shen JJ, See LC, Chang HH, Huang JL . Ding Chuan Tang, a Chinese herb decoction, could improve airway hyper-responsiveness in stabilized asthmatic children: a randomized, double-blind clinical trial[J]. Pediatr Allergy Immunol, 2006,17(5):316-322
[8] Chang TT, Huang CC, Hsu CH . Clinical evaluation of the Chinese herbal medicine formula STA-1 in the treatment of allergic asthma[J]. Phytother Res, 2006,20(5):342-347
doi: 10.1002/(ISSN)1099-1573
[9] Busse PJ, Srivastava K, Wei CH, Li GH, Yu N, Liu YH, Wallenstein S, Wisnivesky J, Qu C, Sampson H, Li XM . Safety, tolerability and beneficial immunological responses of complementary ASHMI therapy to inhaled corticosteroids therapy in children with persistent allergic asthma with or without allergic rhinitis[J]. Allergy Clin Immunol, 2010,125(2):S199
[10] Srivastava K, Zhang T, Yang N, Sampson H, Li XM . Anti-Asthma Simplified Herbal Medicine Intervention-induced long-lasting tolerance to allergen exposure in an asthma model is interferon-gamma, but not transforming growth factor-β dependent[J]. Clin Exp Allergy, 2010,40(11):1678-1688
doi: 10.1111/cea.2010.40.issue-11
[11] Zhang T, Srivastava K, Wen MC, Yang N, Cao J, Busse P, Birmingham N, Goldfarb J, Li XM . Pharmacology and immunological actions of a herbal medicine ASHMI on allergic asthma[J]. Phytother Res, 2010,24(7):1047-1055
[12] Bensky D, Barolet R. Chinese herbal medicine: formulas and strategies. Seattle: Eastland Press. 1990.
[13] Wagner H, Ulrich-Merzenich G . Synergy research: approaching a new generation of phytopharmaceuticals[J]. Phytomedicine, 2009,16(2-3):97-110
doi: 10.1016/j.phymed.2008.12.018
[14] Hosoya E, Yamamura Y . Recent advances in the pharmacology of Kampo(Japanese herbal) medicines[M]. Tokyo: Excerpta Medica, 1998,260
[15] Busse PJ, Schofield B, Birmingham N, Yang N, Wen MC, Zhang T, Srivastava K, Li XM . The traditional Chinese herbal formula ASHMI inhibits allergic lung inflammation in antigen-sensitized and antigen-challenged aged mice[J]. Ann Allergy Asthma Immunol, 2010,104(3):236-246
doi: 10.1016/j.anai.2009.12.006
[16] Kaye J, Porcelli S, Tite J, Jones B, Janeway CA . Both a monoclonal antibody and antisera specific for determinants unique to individual cloned helper T cell lines can substitute for antigen and antigen-presenting cells in the activation of T cells[J]. J Exp Med, 1983,158(3):836-856
doi: 10.1084/jem.158.3.836
[17] Srivastava K, Teper AA, Zhang TF, Li S, Walsh MJ, Huang CK, Kattan M, Schofield BH, Sampson HA, Li XM . Immunomodulatory effect of the anti-asthma Chinese herbal formula, MSSM-002 on Th2 cells[J]. J Allergy Clin Immunol, 2004,113(2):268-276
doi: 10.1016/j.jaci.2003.10.062
[18] Szabo SJ, Kim ST, Costa GL, Zhang X, Fathman CG, Glimcher LH . A novel transcription factor, T-bet, directs Th1 lineage commitment[J]. Cell, 2000,100(6):655-669
doi: 10.1016/S0092-8674(00)80702-3
[19] Conroy DM, Williams TJ . Eotaxin and the attraction of eosinophils to the asthmatic lung[J]. Respir Res, 2001,2(3):150-156
doi: 10.1186/rr52
[20] Sato E, Nelson DK, Koyama S, Hoyt JC, Robbins RA . Inflammatory cytokines modulate eotaxin release by human lung fibroblast cell line[J]. Exp Lung Res, 2001,27(2):173-183
doi: 10.1080/019021401750069401
[21] Jayaprakasam B, Doddaga S, Wang R, Holmes D, Goldfarb J, Li XM . Licorice flavonoids inhibit eotaxin-1 secretion by human fetal lung fibroblasts in vitro[J]. J Agric Food Chem, 2009,57(3):820-825
doi: 10.1021/jf802601j
[22] Matsui S, Matsumoto H, Sonoda Y, Ando K, Aizu-Yokota E, Sato T, Kasahara T . Glycyrrhizin and related compounds down-regulate production of inflammatory chemokines IL-8 and eotaxin 1 in a human lung fibroblast cell line[J]. Int Immunopharmacol, 2004,4(13):1633-1644
doi: 10.1016/j.intimp.2004.07.023
[23] Matsui S, Sonoda Y, Sekiya T, Aizu-Yokota E, Kasahara T . Glycyrrhizin derivative inhibits eotaxin 1 production via STAT6 in human lung fibroblasts[J]. Int Immunopharmacol, 2006,6(3):369-375
doi: 10.1016/j.intimp.2005.08.025
[24] Chinese Pharmacopoeia Commission. Pharmacopoeia of the People’s Republic of China. Beijing: People’s Medical Publishing House. 2005.
[25] Kelly-Pieper K, Patil SP, Busse P, Yang N, Sampson H, Li XM, Wisnivesky JP, Kattan M . Safety and tolerability of an antiasthma herbal formula(ASHMI) in adult subjects with asthma: a randomized, double-blinded, placebo-controlled, dose-escalation phase I study[J]. J Altern Complement Med, 2009,15(7):735-743
doi: 10.1089/acm.2008.0543
[26] U.S.Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research. Guidance for industry: botanical drug products (Revised). 2004.
[27] Li XM, Brown L . Efficacy and mechanisms of action of traditional Chinese medicines for treating asthma and allergy[J]. J Allergy Clin Immunol, 2009,123(2):297-306
doi: 10.1016/j.jaci.2008.12.026
[28] Ministry of Foreign Trade and Economic Cooperation PRC. Green trade standards of importing & exporting medicinal plants & preparations. 2001. Chinese.
[29] Raman P, Patino LC, Nair MG . Evaluation of metal and microbial contamination in botanical supplements[J]. J Agric Food Chem, 2004,52(26):7822-7827
doi: 10.1021/jf049150+
[30] Dolan SP, Nortrup DA, Bolger PM, Capar SG . Analysis of dietary supplements for arsenic, cadmium, mercury, and lead using inductively coupled plasma mass spectrometry[J]. J Agric Food Chem, 2003,51(5):1307-1312
doi: 10.1021/jf026055x
[31] Caldas ED, Machado LL . Cadmium, mercury and lead in medicinal herbs in Brazil[J]. Food Chem Toxicol, 2004,42(4):599-603
doi: 10.1016/j.fct.2003.11.004
[32] Li XM, Schofield BH, Wang QF, Kim KH, Huang SK . Induction of pulmonary allergic responses by antigen-specific Th2 cells[J]. J Immunol, 1998,160(3):1378-1384
[33] Jayaprakasam B, Zhang Y, Seeram NP, Nair MG . Growth inhibition of human tumor cell lines by withanolides from Withania somnifera leaves[J]. Life Sci, 2003,74(1):125-132
doi: 10.1016/j.lfs.2003.07.007
[34] Berenbaum MC . Criteria for analyzing interactions between biologically active agents[J]. Adv Cancer Res, 1981,35:269-335
doi: 10.1016/S0065-230X(08)60912-4
[35] Liu C, Yang N, Folder W, Cohn J, Wang R, Li X . Ganoderic acid C isolated from Ganoderma lucidum suppresses LPS-induced macrophage TNF-α production by down-regulating MAPK, NF-kappaB and AP-1 signaling pathways[J]. J Allergy Clin Immunol, 2012,129(2):S127
[36] Mould AW, Ramsay AJ, Matthaei KI, Young IG, Rothenberg ME, Foster PS . The effect of IL-5 and eotaxin expression in the lung on eosinophil trafficking and degranulation and the induction of bronchial hyperreactivity[J]. J Immunol, 2000,164(4):2142-2150
doi: 10.4049/jimmunol.164.4.2142
[37] Kan WL, Cho CH, Rudd JA, Lin G . Study of the anti-proliferative effects and synergy of phthalides from Angelica sinensis on colon cancer cells[J]. J Ethnopharmacol, 2008,120(1):36-43
doi: 10.1016/j.jep.2008.07.027
[38] Gao JL, He TC, Li YB, Wang YT . A traditional Chinese medicine formulation consisting of Rhizoma Corydalis and Rhizoma Curcumae exerts synergistic anti-tumor activity[J]. Oncol Rep, 2009,22(5):1077-1083
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