ABSTRACT
Column chromatography on silica gel was used to study the chemical constituents of traditional Chinese medicine Siegesbeckia pubescens. The chemical structures of the separated compounds were elucidated by spectroscopic data analyses. As a result, eighteen compounds were obtained and identified as 3, 4'-dimethoxy quercetin(1), 3, 3', 4'-trimethoxy quercetin(2), 3, 3'-dimethoxy quercetin(3), 7, 3', 4'-trimethoxy luteolin(4), ursolic acid(5), 2β,19α-dihydroxyursolic acid(6), 2β-hydroxyursolic acid (7), stigmasterol-7-one(8), 5α, 8α-epidioxy-24(R)-methyl-cholesta-6, 22-diene-3β-ol(9), β-sitosterol(10), 2, 6-di(3-hydroxy-4-methoxyphenyl)-3, 7-dioxacyclo [3. 3. 0] octane (11), aurantiamide acetate (12), 3-(m-hydroxyl-p-methoxy)-N-(2'-p-hydroxyl-phenethyl)-2E-acrylamide(13), p-hydroxy benzaldehyde (14), m-hydroxy-p-methoxy benzaldehyde (15), 3, 4, 5-trimethoxybenzoic acid(16), monoethyl malonate(17), and p-hydroxylcinnamic acid(18). Among them, compounds 1-9, 11-18 were isolated from this plant for the first time.
Subject(s)
Asteraceae , Chemistry , Drugs, Chinese Herbal , Chemistry , Luteolin , Chemistry , Medicine, Chinese Traditional , Plants, Medicinal , Quercetin , Chemistry , Sitosterols , Chemistry , Triterpenes , ChemistryABSTRACT
<p><b>AIM</b>To establish an in vitro long-term culture system of mouse Spermatogonial stem cells (SSCs).</p><p><b>METHODS</b>Three types of serum-free culture media, namely, DMEM/F12, KSR (KnockoutM Serum Replacement) and StemPro-34 SFM, to which the same growth factors including GDNF, soluble GFRalpha1 and bFGF were added equally, and MEF(mouse embryonic fibroblast) feeder layer were used to culture mouse SSCs enriched from pup mice testes through differential adherence selection. The activity of stem cells was examined morphologically, and the marker gene expression of SSCs was detected by RT-PCR and immunocytochemical analysis.</p><p><b>RESULTS</b>The activity of SSCs cultured in DMEM/F12 and KSR serum-free media was only maintained for 6-7 days. However, the StemPro-34 SFM medium could maintain the proliferation of cultured SSCs nearly one month.</p><p><b>CONCLUSION</b>StemPro-34 SFM serum-free medium sustains the proliferation of mouse SSCs in vitro.</p>