Open AccessSemi‐quantitative analysis of cytokine mRNA expression induced by the herbal medicine Sho‐saiko‐to (TJ‐9) using a Gel Doc system
Author(s) -
Huang XianXi,
Yamashiki Masayoshi,
Nakatani Kaname,
Nobori Tsutomu,
Mase Akihito
Publication year - 2001
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1028
Subject(s) - messenger rna , cytokine , traditional medicine , medicine , computational biology , pharmacology , immunology , chemistry , biology , biochemistry , gene
The RT‐PCR method was employed to determine the cytokine mRNA expression of human peripheral lymphocytes induced by the Japanese herbal medicine Sho‐saiko‐to (TJ‐9). The results showed that the mRNA expression of IL‐12, IL‐1β, IL‐10, TNF‐α, G‐CSF, and IFN‐γ increased after 6 hr in culture. This is the first reported finding that TJ‐9 is an IFN‐γ inducer. Next, cytokine mRNA expression was semi‐quantitatively measured using the Gel Doc system with a CCD camera and then statistically analyzed in order to determine which component of TJ‐9 was the true cytokine inducer. The results showed that the scutellaria root is the main component inducing the cytokines, while the glycyrrhiza root is the secondary component. When the cytokine concentrations in the supernatants of cell cultures were measured by ELISA, the levels of IL‐12, IL‐1β, IL‐10, TNF‐α, and G‐CSF reflected mRNA expression levels in the cell fraction. However, the level of IFN‐γ was below the detectable limit. The effects of various reagents on many different kinds of cytokine mRNA expression could be analyzed objectively in a short time using the Gel Doc system. Many important findings could be demonstrated by this simple, easy, sensitive, and cheap method. After the clinical significance of cytokine analysis is confirmed, this method may become a useful clinical examination tool. J. Clin. Lab. Anal. 15:199–209, 2001. © 2001 Wiley‐Liss, Inc.