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Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (4): 341-346.doi: 10.3736/jcim20100407

• Original Experimental Research • Previous Articles     Next Articles

Effects of Cordyceps extract on cytokines and transcription factors in peripheral blood mononuclear cells of asthmatic children during remission stage

 Wen Suna, Jian Yua, Yu-min Shia, Hao Zhangb, Ying Wangc, Bing-bing Wuc   

  1. a Department of Traditional Chinese Medicine,Children’s Hospital, Fudan University, Shanghai 201102, China;
    b Department of Respiratory Diseases, Children’s Hospital, Fudan University, Shanghai 201102, China;
    c Pediatric Institute, Children’s Hospital, Fudan University, Shanghai 201102, China;
  • Received:2009-11-18 Accepted:2010-02-08 Online:2010-04-20 Published:2010-04-15
  • Contact: Jian Yu E-mail:yuj20061966@yahoo.com.cn

Objective:To explore the effects of Cordyceps extract in regulating the imbalance of Th1/Th2 ratio and inhibiting the inflammatory reaction, and to find the theoretical basis of Cordyceps extract for treating asthma in remission stage.
Methods:A total of 20 peripheral venous blood samples (3 mL) were collected from 20 asthmatic children during remission stage, and peripheral blood mononuclear cells (PBMCs) were separated by Ficoll method. PBMCs were separated into three groups (blank group, low-dose group and high-dose group). The PBMCs were incubated in vitro for 48 hours in the absence (blank group) or presence (low-dose group and high-dose group) of Cordyceps extract at different concentrations (10, 20 μg/mL). The expressions of interferon-γ (IFN-γ), interleukin-4 (IL-4), IL-10, T-box expressed in T cells (T-bet), GATA-binding protein-3 (GATA-3) and forkhead/winged-helix transcription factor-3 (Foxp3) mRNAs in PBMCs were measured by real-time fluorescent quantitative polymerase chain reaction, and the contents of IFN-γ, IL-4 and IL-10 in supernatants were measured by enzyme-linked immunosorbent assay.
Results:The expressions of IFN-γ mRNA showed no significant differences among the three groups. The expressions of the IL-4 mRNA in the high-dose group and the low-dose group were lower than that in the blank group (P=0.014, P=0.011). The expression of IL-10 mRNA in the high-dose group was higher than that in the blank group (P=0.034). And the differences of the IFN-γ mRNA/IL-4 mRNA ratio presented no statistic significance among the three groups. The level of IL-4 content in the high-dose group was lower than that in the blank group (P=0.018), but the level of IL-10 content, and ratio of IFN-γ/IL-4 in the high-dose group were higher than those in the blank group (P=0.011, P=0.045). The differences of the IFN-γ presented no statistic significance among the three groups. The T-bet/GATA-3 ratio and Foxp3 mRNA expression in the high-dose group were higher than those in the blank group (P=0.001, P=0.015). There was significant difference in expression of GATA-3 mRNA between the high-dose group and the blank group (P=0.028), and between the low-dose group and the blank group (P=0.019). The expression differences of T-bet mRNA were insignificant between any two groups.
Conclusion:Cordyceps extract can inhibit the proliferation and differentiation of Th2 cells and reduce the expression of related cytokines by down-regulating the expression of GATA-3 mRNA and up-regulating the expression of Foxp3 mRNA in PBMCs. Meanwhile, it can alleviate the chronic allergic inflammation by increasing the content of IL-10.

Key words: Cordyceps sinensis, Asthma, Monocytes, Remission stage, Cytokines, Transcription factors

Table 1

Sequence of primers and length of the products"

Gene Sequence of primer (5′→3′) Length of product (bp)
β-actin Upstream: CAC CAA CTG GGA CGA CAT 138
Downstream:ATC TGG GTC ATC TTC TCG C
IFN-γ Upstream: CCA ACG CAA AGC AAT ACA 133
Downstream: CAG GCA GGA CAA CCA TTA
IL-4 Upstream: ACC GAG TTG ACC GTA ACA GAC 151
Downstream: TGT GGA ACT GCT GTG CAG T
IL-10 Upstream: CCA AGC TGA GAA CCA AGA CC 152
Downstream: TTA AGG CAT TCT TCA CCT GCT
T-bet Upstream: CCG GCT GCA TAT CGT TGA 132
Downstream: TGA GTA ATC TCG GCA TTC TGG
GATA-3 Upstream: GAC AGA ACC GGC CCC TCA TTA 221
Downstream: CAT TTT TCG GTT TCT GGT CTG GAT
Foxp3 Upstream: TGG TGC TGG AGA AGG AGA A 298
Downstream: AGC GTG GCG TAG GTG AAA

Table 2

Expressions of IL-4, IFN-γ and IL-10 mRNAs in PBMCs in different groups ($\bar{x}$±s)"

Group n IFN-γ mRNA IL-4 mRNA IFN-γ mRNA/IL-4 mRNA IL-10 mRNA
Blank 20 1.49±0.52 1.11±0.47 1.56±0.78 2.66±0.38
Low-dose Cordyceps extract 20 1.47±0.52 0.78±0.35* 2.23±0.75 2.79±0.44
High-dose Cordyceps extract 20 1.50±0.47 0.76±0.41* 2.20±0.91 2.98±0.44*

Table 3

Contents of IL-4, IFN-γ and IL-10 in supernatant in different groups ($\bar{x}$±s, ng/L)"

Group n IFN-γ IL-4 IFN-γ/IL-4 IL-10
Blank 20 130.37±12.17 48.97±3.17 2.75±0.95 237.20±18.80
Low-dose Cordyceps extract 20 133.41±8.23 42.07±2.60 3.40±1.14 306.11±29.71
High-dose Cordyceps extract 20 134.74±8.61 38.49±2.47* 3.70±1.05* 328.11±32.83*

Table 4

Expressions of T-bet, GATA-3 and Foxp3 mRNAs in PBMCs in different groups ($\bar{x}$±s)"

Group n T-bet mRNA GATA-3 mRNA T-bet mRNA/GATA-3 mRNA Foxp3 mRNA
Blank 20 2.94±0.23 2.59±0.21 1.16±0.05 2.31±0.30
Low-dose Cordyceps extract 20 2.83±0.21 1.97±0.14* 1.46±0.07** 2.50±0.26
High-dose Cordyceps extract 20 2.96±0.22 2.01±0.13* 1.50±0.08** 2.56±0.29*
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