Isolation and identification of hematopoietic stem cell-stimulating substances from Kampo (Japanese herbal) medicine, Juzen-taiho-to.
We have previously found that TJ-48 has the capacity to accelerate recovery from hematopoietic injury induced by radiation and the anti-cancer drug mitomycin C (MMC). The effects are found to be due to its stimulation of spleen colony-forming unit (CFU-S) counts on day 14. In the present study, we attempt to isolate and purify the active components in TJ-48 extracts using a new in vitro hematopoietic stem cell (HSC) assay method. n-Hexane extract from TJ-48 shows a significant stimulatory activity. The extract is further fractionated by silica gel chromatography and HPLC in order to identify its active components. 1H-NMR and GC-EI-MS indicate that the active fraction is composed of free fatty acids (oleic acid and linolenic acid). When 27 kinds of free fatty acids (commercially available) are tested using the HSC proliferating assay, oleic acid, elaidic acid, and linolenic acid are found to have potent activity. The administration of oleic acid to MMC-treated mice enhances CFU-S counts on days 8 and 14 to twice the control group. These findings strongly suggest that fatty acids contained in TJ-48 actively promote the proliferation of HSCs. Although many mechanisms seem to be involved in the stimulation of HSC proliferation, we speculate that at least one of the signals is mediated by stromal cells, rather than any direct interaction with the HSCs.
Hisha
H
H
1st Department of Pathology, Kansai Medical University, Moriguchi City, Osaka, Japan.
Yamada
H
H
Sakurai
M H
MH
Kiyohara
H
H
Li
Y
Y
Yu
C
C
Takemoto
N
N
Kawamura
H
H
Yamaura
K
K
Shinohara
S
S
Komatsu
Y
Y
Aburada
M
M
Ikehara
S
S
D000818:Animals; D001853:Bone Marrow / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D005591:Chemical Fractionation / Q000379:methods; D002725:Chloroform; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003114:Colony-Forming Units Assay; D003238:Connective Tissue / Q000187:drug effects; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D005227:Fatty Acids / Q000302:isolation & purification* / Q000494:pharmacology; D008401:Gas Chromatography-Mass Spectrometry; D006410:Hematopoiesis / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D006586:Hexanes; D009682:Magnetic Resonance Spectroscopy; D000432:Methanol; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008934:Mitogens / Q000302:isolation & purification*; D016685:Mitomycin / Q000633:toxicity; D019301:Oleic Acid / Q000302:isolation & purification / Q000494:pharmacology; D009829:Oleic Acids; D012997:Solvents; D017962:alpha-Linolenic Acid / Q000302:isolation & purification / Q000494:pharmacology
D004365:Drugs, Chinese Herbal; D005227:Fatty Acids; D006586:Hexanes; D008934:Mitogens; D009829:Oleic Acids; D012997:Solvents; C063587:juzentaihoto; D017962:alpha-Linolenic Acid; C026385:n-hexane; D019301:Oleic Acid; C011459:elaidic acid; D016685:Mitomycin; D002725:Chloroform; D000432:Methanol