title: The Kampo medicines Orengedokuto, Bofutsushosan and Boiogito have different activities to regulate gene expressions in differentiated rat white adipocytes: comprehensive analysis of genetic profiles.
abstract: Three Kampo medicines, Boiogito (BOT), Bofutsushosan (BTS) and Orengedokuto (OGT), used for obese patients were investigated for their effects on adipogenesis in cultured rat white adipocytes. Administration of the three extracts suppressed adipogenesis in concentration-dependent manners (1-100 microg/ml) without any cytotoxicity. Changes in mRNA expression levels were analyzed using a Rat 230 2.0 Affymetrix GeneChip microarray system. DNA microarray analysis (total probe set: 31099) using cDNAs prepared from adipocytes revealed that BOT, BTS and OGT increased the expression of 133-150 genes and decreased the expression of 42-110 genes by > or =2-fold. We identified 329 downregulated genes and 189 upregulated genes among a total set of 514 probes (overlap: 4). Overall, genes related to cellular movement, cell death, cell growth/differentiation and immune responses were the most downregulated, while those related to lipid metabolism and cell signaling were the most upregulated. Semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assays were conducted to confirm the microarray results. Analysis of the clustering profiles of the microarray results revealed that BOT and BTS changed the expression levels of similar genes mainly involved in small molecule biochemistry and cell differentiation, while OGT altered 10 genes related to lipid metabolism, in contrast to the effects of BOT and BTS. We also measured mRNA expression levels of seven selected genes highly contributing to the lipid metabolism by using semiquantitative RT-PCR assay, that were acetyl-Coenzyme A carboxylase alpha (ACACA), AE binding protein 1 (AEBP1), patatin-like phospholipase domain containing 8 (PNPLA8), secretoglobin (SCGB1A1), adrenergic (ADRB3), adiponectin (ADIPOQ), monoglyceride lipase (MGLL). Beta-actin (ACTB) gene was used as an endogenous internal standard. The present findings indicate that these three herbal extracts have the potential to prevent adipogenesis in rat white adipocytes through different mechanisms via modulation of gene expression levels.
authors.affiliation: Department of General Medicine, Kanazawa Medical University, Japan.
authors.forename: Jun-ichi
authors.initials: J
authors.lastname: Yamakawa
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Ishigaki
authors.forename: Fumihide
authors.initials: F
authors.lastname: Takano
authors.forename: Takashi
authors.initials: T
authors.lastname: Takahashi
authors.forename: Junko
authors.initials: J
authors.lastname: Yoshida
authors.forename: Junji
authors.initials: J
authors.lastname: Moriya
authors.forename: Takanobu
authors.initials: T
authors.lastname: Takata
authors.forename: Takanori
authors.initials: T
authors.lastname: Tatsuno
authors.forename: Kenroh
authors.initials: K
authors.lastname: Sasaki
authors.forename: Tomihisa
authors.initials: T
authors.lastname: Ohta
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Takegami
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Yoshizaki
mesh_terms: D052438:Adipocytes, White / Q000187:drug effects* / Q000378:metabolism; D050156:Adipogenesis / Q000187:drug effects* / Q000235:genetics; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D002478:Cells, Cultured; D015536:Down-Regulation; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D020835:Medicine, Kampo; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
chemical_list: D004365:Drugs, Chinese Herbal; C098310:bofu-tsusho-san; C523704:boiogito; C061676:oren gedoku to