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Use of serial analysis of gene expression to reveal the specific regulation of gene expression profile in asthmatic rats treated by acupuncture.

Yin LM, Jiang GH, Wang Y, Wang Y, Liu YY, Jin WR, Xu YD, Zhang QH, Yang YQ - J. Biomed. Sci. (2009)

Bottom Line: By comparison of P(CK/AS) and P(AS/ASAC), the DAVID genes functional classification was found to be changed from "immune response" to "response to steroid hormone stimulus", and the GO term "antigen processing and presentation of peptide antigen" disappeared in P(AS/ASAC).Totally 3 same KEGG pathways were found among the three groups.Our SAGE research indicates that the gene expression profile of the EAR phase of asthma could be effectively and specifically regulated by acupuncture, which suggests that the gene expression of immune response and steroid hormone may play an important role in the treatment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Shanghai Research Institute of Acupuncture and Meridian, Shanghai University of Traditional Chinese Medicine, Shanghai, PR China. collegeylm@hotmail.com

ABSTRACT

Background: Asthma has become an important public health issue and approximately 300 million people have suffered from the disease worldwide. Nowadays, the use of acupuncture in asthma is increasing. This study intended to systematically analyze and compare the gene expression profiles between the asthmatic and acupuncture-treated asthmatic rat lung, and tried to gain insight into the molecular mechanism underlying the early airway response (EAR) phase of asthma treated by acupuncture.

Methods: Four tag libraries of serial analysis of gene expression (SAGE) were established from lung tissues of control rats (CK), asthmatic rats (AS), asthmatic rats treated by acupuncture (ASAC), and control rats treated by acupuncture (CKAC). Bioinformatic analyses were carried out by using the methods including unsupervised hierarchical clustering, functional annotation tool of the database for annotation, visualization, and integrated discovery (DAVID), gene ontology (GO) tree machine, and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis.

Results: There were totally 186 differentially expressed tags (P < 0.05, P(CK/AS)) between the libraries of CK and AS, 130 differentially expressed tags between libraries of AS/ASAC (P < 0.05, P(AS/ASAC)), and 144 differentially expressed tags between libraries of CK/CKAC (P < 0.05, P(CK/CKAC)). The gene expression profiles of AS and ASAC were more similar than other libraries via unsupervised SAGE clustering. By comparison of P(CK/AS) and P(AS/ASAC), the DAVID genes functional classification was found to be changed from "immune response" to "response to steroid hormone stimulus", and the GO term "antigen processing and presentation of peptide antigen" disappeared in P(AS/ASAC). Totally 3 same KEGG pathways were found among the three groups. Moreover, 21 specific tags of the acupuncture in treating asthma were detected using Venn diagrams.

Conclusion: Our SAGE research indicates that the gene expression profile of the EAR phase of asthma could be effectively and specifically regulated by acupuncture, which suggests that the gene expression of immune response and steroid hormone may play an important role in the treatment.

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Related in: MedlinePlus

Protocol and measurements for the rat model of allergic asthma. Male Sprague-Dawley rats were sensitized intraperitoneally at day 0 with 1 mg of ovalbumin (OVA) precipitated with 10 mg of aluminium hydroxide gel in 1 ml normal saline. Manual acupuncture was performed regularly from the first day after the sensitization and given every other day over a period of two weeks. On day 14, the sensitized animals were challenged with 1 ml/kg of 5% OVA in normal saline (5 mg/kg) by injection into the external jugular vein over 10 s. The respiratory parameters of pulmonary resistance, dynamic compliance and respiratory rate were immediately recorded for 10 min. Rats were killed immediately after the measurement. Rats of CK and CKAC were sensitized and challenged with normal saline instead of OVA. Rats of CKAC received the same acupuncture treatment as those of ASAC.
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Figure 1: Protocol and measurements for the rat model of allergic asthma. Male Sprague-Dawley rats were sensitized intraperitoneally at day 0 with 1 mg of ovalbumin (OVA) precipitated with 10 mg of aluminium hydroxide gel in 1 ml normal saline. Manual acupuncture was performed regularly from the first day after the sensitization and given every other day over a period of two weeks. On day 14, the sensitized animals were challenged with 1 ml/kg of 5% OVA in normal saline (5 mg/kg) by injection into the external jugular vein over 10 s. The respiratory parameters of pulmonary resistance, dynamic compliance and respiratory rate were immediately recorded for 10 min. Rats were killed immediately after the measurement. Rats of CK and CKAC were sensitized and challenged with normal saline instead of OVA. Rats of CKAC received the same acupuncture treatment as those of ASAC.

Mentions: Manual acupuncture was performed for two weeks from the first day after the sensitization, once every other day (Figure 1). The acupuncture points, Dazhui (GV 14, between C7 and T1 vertebrae), bilateral Fengmen (BL12, foveola laterally between T2 and T3 vertebrae) and bilateral Feishu (BL13, foveola laterally between T3 and T4 vertebrae), were selected based on the theory of TCM in treating asthma [22]. In consideration of the specific and non-specific effects of acupuncture [23], the same points in normal rats were selected and acted as control of ASAC, which could distinguish the specificity of acupuncture in treating asthma. The stainless needles (0.30 × 13 mm, Suzhou Medical Appliance Factory, Suzhou, China) were inserted about 5 mm deep into the skin. The needles were twisted approximately 360° evenly at the rate of 60 times/min for 20 s, manipulated every 5 min and withdrawn after 20 min. For the convenient manipulation of the acupuncture points on the back, rat was placed on the suspended shelf (50 × 45 mm, about 50 cm high from the ground, see additional file 1), which could easily make it calm and stand still without anesthesia. Acupuncture was performed by the same experienced practitioner and the animals were handled while awake, with special care to minimize stress. Both rats in ASAC and CKAC received the same acupuncture treatment.


Use of serial analysis of gene expression to reveal the specific regulation of gene expression profile in asthmatic rats treated by acupuncture.

Yin LM, Jiang GH, Wang Y, Wang Y, Liu YY, Jin WR, Xu YD, Zhang QH, Yang YQ - J. Biomed. Sci. (2009)

Protocol and measurements for the rat model of allergic asthma. Male Sprague-Dawley rats were sensitized intraperitoneally at day 0 with 1 mg of ovalbumin (OVA) precipitated with 10 mg of aluminium hydroxide gel in 1 ml normal saline. Manual acupuncture was performed regularly from the first day after the sensitization and given every other day over a period of two weeks. On day 14, the sensitized animals were challenged with 1 ml/kg of 5% OVA in normal saline (5 mg/kg) by injection into the external jugular vein over 10 s. The respiratory parameters of pulmonary resistance, dynamic compliance and respiratory rate were immediately recorded for 10 min. Rats were killed immediately after the measurement. Rats of CK and CKAC were sensitized and challenged with normal saline instead of OVA. Rats of CKAC received the same acupuncture treatment as those of ASAC.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2698896&req=5

Figure 1: Protocol and measurements for the rat model of allergic asthma. Male Sprague-Dawley rats were sensitized intraperitoneally at day 0 with 1 mg of ovalbumin (OVA) precipitated with 10 mg of aluminium hydroxide gel in 1 ml normal saline. Manual acupuncture was performed regularly from the first day after the sensitization and given every other day over a period of two weeks. On day 14, the sensitized animals were challenged with 1 ml/kg of 5% OVA in normal saline (5 mg/kg) by injection into the external jugular vein over 10 s. The respiratory parameters of pulmonary resistance, dynamic compliance and respiratory rate were immediately recorded for 10 min. Rats were killed immediately after the measurement. Rats of CK and CKAC were sensitized and challenged with normal saline instead of OVA. Rats of CKAC received the same acupuncture treatment as those of ASAC.
Mentions: Manual acupuncture was performed for two weeks from the first day after the sensitization, once every other day (Figure 1). The acupuncture points, Dazhui (GV 14, between C7 and T1 vertebrae), bilateral Fengmen (BL12, foveola laterally between T2 and T3 vertebrae) and bilateral Feishu (BL13, foveola laterally between T3 and T4 vertebrae), were selected based on the theory of TCM in treating asthma [22]. In consideration of the specific and non-specific effects of acupuncture [23], the same points in normal rats were selected and acted as control of ASAC, which could distinguish the specificity of acupuncture in treating asthma. The stainless needles (0.30 × 13 mm, Suzhou Medical Appliance Factory, Suzhou, China) were inserted about 5 mm deep into the skin. The needles were twisted approximately 360° evenly at the rate of 60 times/min for 20 s, manipulated every 5 min and withdrawn after 20 min. For the convenient manipulation of the acupuncture points on the back, rat was placed on the suspended shelf (50 × 45 mm, about 50 cm high from the ground, see additional file 1), which could easily make it calm and stand still without anesthesia. Acupuncture was performed by the same experienced practitioner and the animals were handled while awake, with special care to minimize stress. Both rats in ASAC and CKAC received the same acupuncture treatment.

Bottom Line: By comparison of P(CK/AS) and P(AS/ASAC), the DAVID genes functional classification was found to be changed from "immune response" to "response to steroid hormone stimulus", and the GO term "antigen processing and presentation of peptide antigen" disappeared in P(AS/ASAC).Totally 3 same KEGG pathways were found among the three groups.Our SAGE research indicates that the gene expression profile of the EAR phase of asthma could be effectively and specifically regulated by acupuncture, which suggests that the gene expression of immune response and steroid hormone may play an important role in the treatment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Shanghai Research Institute of Acupuncture and Meridian, Shanghai University of Traditional Chinese Medicine, Shanghai, PR China. collegeylm@hotmail.com

ABSTRACT

Background: Asthma has become an important public health issue and approximately 300 million people have suffered from the disease worldwide. Nowadays, the use of acupuncture in asthma is increasing. This study intended to systematically analyze and compare the gene expression profiles between the asthmatic and acupuncture-treated asthmatic rat lung, and tried to gain insight into the molecular mechanism underlying the early airway response (EAR) phase of asthma treated by acupuncture.

Methods: Four tag libraries of serial analysis of gene expression (SAGE) were established from lung tissues of control rats (CK), asthmatic rats (AS), asthmatic rats treated by acupuncture (ASAC), and control rats treated by acupuncture (CKAC). Bioinformatic analyses were carried out by using the methods including unsupervised hierarchical clustering, functional annotation tool of the database for annotation, visualization, and integrated discovery (DAVID), gene ontology (GO) tree machine, and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis.

Results: There were totally 186 differentially expressed tags (P < 0.05, P(CK/AS)) between the libraries of CK and AS, 130 differentially expressed tags between libraries of AS/ASAC (P < 0.05, P(AS/ASAC)), and 144 differentially expressed tags between libraries of CK/CKAC (P < 0.05, P(CK/CKAC)). The gene expression profiles of AS and ASAC were more similar than other libraries via unsupervised SAGE clustering. By comparison of P(CK/AS) and P(AS/ASAC), the DAVID genes functional classification was found to be changed from "immune response" to "response to steroid hormone stimulus", and the GO term "antigen processing and presentation of peptide antigen" disappeared in P(AS/ASAC). Totally 3 same KEGG pathways were found among the three groups. Moreover, 21 specific tags of the acupuncture in treating asthma were detected using Venn diagrams.

Conclusion: Our SAGE research indicates that the gene expression profile of the EAR phase of asthma could be effectively and specifically regulated by acupuncture, which suggests that the gene expression of immune response and steroid hormone may play an important role in the treatment.

Show MeSH
Related in: MedlinePlus