JBM  Vol.3 No.7 , July 2015
Establishment of Cytokines Quantitative PCR Methods and Its Application in Anaphylaxis Study of Houttuynia Cordata Injection
ABSTRACT
We design specific primers, according to the four kinds of important macaque cytokines, based on macaque β-action genes, then extract total RNA in peripheral blood mononuclear cells of macaques, and trascriptase cDNA reversely after establishing real-time quantitative PCR, and test the relative expression amount of several kinds of important cytokines during the hypersensitive process. The testing method of quantitative PCR in this experiment is highly sensitive, all cytokines are up to 102 copies/μL and highly reproducible, the intraassay and interassay coefficients of variation are less than 6%. The method applied to the sensitive reaction experiment on cytokines induced by houttuynia cordata turns out that the cytokines relative expression of houttuynia cordata distillate with low dose group and high dose group is significantly higher than that of saline group (P > 0.01). Meanwhile, significantly increased IL-4, IL-10 and IL-13 and decreased IFN-γ are found in both group A and group B after houttuynia injection (P > 0.05).

Cite this paper
Zeng, X. , Sun, F. , Cheng, X. , Zeng, W. and Li, L. (2015) Establishment of Cytokines Quantitative PCR Methods and Its Application in Anaphylaxis Study of Houttuynia Cordata Injection. Journal of Biosciences and Medicines, 3, 1-8. doi: 10.4236/jbm.2015.37001.
References
[1]   Varin, A., Mukhopadhyay, S., Herbein, G. and Gordon, S. (2010) Alternative Activation of Macrophages by IL-4 Impairs Phagocytosis of Pathogens but Potentiates Microbial-Induced Signaling and Cytokine Secretion. Blood, 115, 353- 362.
http://dx.doi.org/10.1182/blood-2009-08-236711

[2]   Kai, F., Justine, T., Bernhard, O., Lehmann, P.O. and Tary-Lehmann, M. (2010) Th1 Responses to Beta-Amploid in Young Humans Convert to Regulatory IL-10 Responses in Down Syndrome and Alzheimer’s Disease. Neurobiology of Aging, 31, 1732-1742.
http://dx.doi.org/10.1016/j.neurobiolaging.2008.09.007

[3]   Yang, M., Cheng, A.C., Wang, M.S. and Xing, H.Y. (2008) Development and Application of One-Step Real-Time Taqman RT-PCR Assay for Detection of Duck Hepatitis Virus Type1. Journal of Virological Methods, 153, 55-60.
http://dx.doi.org/10.1016/j.jviromet.2008.06.012

[4]   de Coaña, Y.P., Barrero, C., Cajiao, I., Mosquera, C., Patarroyo, M.E. and Patarroyo, M.A. (2004) Quantifying Aotus Monkey Cytokines by Real-Time Quantitative RT-PCR. Cytokine, 27, 129-133.
http://dx.doi.org/10.1016/j.cyto.2004.04.004

[5]   Stordeur, P., Poulin, L., Craciun, L., Zhou, L., Schandené, L., de Lavareille, A., et al. (2002) Cytokine mRNA Quantification by Real-Time PCR. Journal of Immunological Method, 259, 552-641.
http://dx.doi.org/10.1016/s0022-1759(01)00489-6

[6]   Giulietti, A., Overbergh, L., Valckx, D., Brigitte Decallonne, B., Roger Bouillon, R. and Chantal Mathieu, C. (2001) An Overview of Real-Time Quantitative PCR: Applications to Quantify Cytokines Gene Expression. Methods, 25, 386-401.
http://dx.doi.org/10.1006/meth.2001.1261

[7]   Rudge, E.U., Cutler, A.J., Pritchard, N.R. and Smith, K.G.C. (2002) Interleukin 4 Reduces Expression of Inhibitory receptors on B Cells and Abolishes CD22 and FcγRII-Mediated B Cell Suppression. The Journal of Experimental Medicine, 195, 1079-1085.
http://dx.doi.org/10.1084/jem.20011435

[8]   Pawakar, R., Okuda, M., Yssel, H., Okumura, K. and Ra, C. (1997) Nasal Mast cells in Perennial Allergic Rhinitics Exhibit Increased Expression of the FcεRI, CD40L, IL-4, and IL-13, and Can Induce IgE Synthesis in B Cells. Journal of Clinical Investigation, 99, 1492-1499.
http://dx.doi.org/10.1172/JCI119311

[9]   Wills-Karp, M., Luyimbazi, J., Xu, X.Y., Schofield, B., Neben, T.Y., Karp, C.L., et al. (1998) Interlukin-13: Central Mediator of Allergic Asthma. Science, 282, 2258-2261.
http://dx.doi.org/10.1126/science.282.5397.2258

[10]   Wong, C.K., Ho, C.Y. Ko, W.S., Chan, C.H.S., Ho, A.S.S. Hui, D.S.C. and Lam, C.W.K. (2001) Proinflammatory Cytokines (IL-17, IL-6, IL-18 and IL-12) and Th Cytokines (IFN-γ, IL-4, IL-10 and IL-13) in Patients with Allergic Asthma. Clinical and Experimental Immunology, 125, 177-183.
http://dx.doi.org/10.1046/j.1365-2249.2001.01602.x

 
 
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