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Bioactivity‐guided isolation and identification of anti‐adipogenic compounds from Sanguisorba officinalis L.
Author(s) -
Wang Zhiqiang,
Im Sun Hyuk,
Lim Soon Sung,
Lee OkHwan,
Kang IlJun
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.lb358
Subject(s) - chemistry , officinalis , chromatography , sterol , ellagic acid , ethyl acetate , fractionation , biochemistry , polyphenol , biology , botany , cholesterol , antioxidant
Sanguisorba officinalis L. is a medicinal plant used traditionally for the treatment of inflammatory and metabolic diseases in Korea, China, and Japan. In our previous study, its 50% ethanol extract showed inhibitory effects on triglyceride accumulation through down‐regulation of the sterol regulatory element‐binding protein‐1c (SREBP‐1c), peroxisome proliferators‐activator receptor‐γ (PPAR‐γ), and CCAAT/enhancer binding protein‐α (C/EBP‐α) in 3T3‐L1 adipocytes. Thus, the aim of this study was to investigate bioassay guided fractionation, isolation, and identification of anti‐adipogenic bioactive compounds in S. officinalis L. by using effective differentiation of 3T3‐L1 cells to adipocytes. S. officinalis L. (1 kg) was extracted using 50% ethanol, and the freeze‐dried extract powder (279 g) was partitioned successively using n ‐hexane ( n ‐Hex), methylene chloride (MC), ethyl acetate (EA), n ‐butanol ( n ‐BuOH), and water (Wat) to yield n ‐Hex fraction (4.74 g), MC fraction (5.85 g), EA fraction (18.14 g), n ‐BuOH fraction (42.96 g), and Wat fraction (167.45 g). EA fraction showed significant inhibition of lipid accumulation (44.84%) at the concentration of 50 μg/mL, without cytotoxicity. Activity‐guided isolation using column chromatography from the EA fraction enabled precise localization of the compounds responsible for the anti‐differentiation activity of 3T3‐L1 cells to adipocytes, which were isolated and identified as the known compounds isorhamnetin‐3‐ O ‐D‐glucuronide ( 1 ) and ellagic acid ( 2 ). These results suggested that S. officinalis L. is a potential natural ingredient for the prevention of obesity, which may due to bioactive compounds such as ellagic acid and isorhamnetin‐3‐ O ‐D‐glucuronide. 1Extraction and fractionation procedures of Sanguisorba officinalis L.2 Effect of solvent fractions of Sanguisorba officinalis L. extracts on lipid accumulation in 3T3‐L1 adipocytes(A) The photographed after Oil Red O staining. (B) Stained lipids were extracted and quantified by measuring absorbance at 570 nm. Each value is expressed as the mean ± S.D. Values with different superscripts are significantly different at p <0.05.3HPLC chromatograms of compounds 1 and 2 of ethyl acetate fraction. (A) Ethyl acetate fraction of Sanguisorba officinalis L. 50% EtOH extract, (B) compound 1 (Isorhamnetin 3‐ O ‐glucuronide), (C) compound 2 (Ellagic acid). The HPLC equipment was Gilson TRILUTION 2.1 with UV detector (254 and 362 nm).4Effect of compound 1 (Isorhamnetin 3‐ O ‐glucuronide)on lipid accumulation in 3T3‐L1 adipocytes. (A) The morphological changes associated with cell differentiation were photographed after Oil Red O staining. (B) Stained lipids were extracted and quantified by measuring absorbance at 570 nm. Each value is expressed as the mean ± S.D. Values with different superscripts are significantly different at p <0.05. Control: undifferentiated preadipocyte; MDI: differentiated adipocyte.