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Study on the Separation and Purification Way of Arctiin from Fructus Arctii and the Effect of Arctiin on the Growth of Mice C2C12 Cells

Author: LiuZe
Tutor: GaoFeng
School: Nanjing Agricultural College
Course: Animal Nutrition and Feed Science
Keywords: Frucuts arctii Arctiin extraction purification growth estrogen receptorβ
CLC: S853.7
Type: Master's thesis
Year: 2011
Downloads: 7
Quote: 0
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Abstract


Frucuts arctii is the dried seeds of Arctium lappa L. (Compositae), which has a wide distribution and a long medicinal history. Arctiin is a substance with a variety of physiological activities. Frucuts arctii was taken as the object of study in this topic, its effective component Arctiin was extracted, purified, and its biological activity was preliminarily researched, and these studies may set a foundation for the further development of Frucuts arctii lignan, Arctiin.Firstly, the extraction plan of Arctiin from Frucuts arctii was determined. Taking Arctiin as the standard reference substance compounds, the content of Arctiin was determined by HPLC. Taking yield rate of Arctiin as index, the orthogonal experiment were carried on to obtain the optimization extraction craft parameter, finally the extraction plan of Arctiin was determined:ethanol concentration 70%(V/V), solid-liquid ratio 1:8(W/V), extraction time 1h, and 3 times of extraction.Secondly, Arctiin from Frucuts arctii were separated by macroporous absorption resin column chromatograph and silica gel column chromatograph, elution process was detected by TLC. Taking Arctiin as the standard reference substance compounds, the content of Arctiin was determined by HPLC. Eventually, Arctiin were obtained and the content≥80%. At the same time, we separated the D4006, the one with the maximal binding ability for Arctiin, from six types of resins. The optimum purification conditions were as follows:the adsorption velocity was 2BV/h, the ratio of diameter to height of resin column was 1:5, 50% alcohol was chosen as elution agent,4 times as the volume of the resin. And,50% alcohol lotion was collected and dried. The content of Arctiin was tested≥80%.Finally, the growth-promoting activities of flaxseed lignans was preliminarily studied All experiments were conducted on C2C12 myoblasts (24 h after passaging) and differentiating myoblasts (96 h after induction of fusion). After 24 hours’ incubation, C2C12 myoblasts proliferation were assessed by MTT activity. After 48 hours’ incubation, differentiating myoblasts’ total protein, CK activity and the expression of estrogen receptorβ(ERβ) were investigated by Coomassie brilliant blue method, colorimetric analysis and QRT-PCR, respectively. The result showed that C2C12 myoblasts’ proliferation was significantly improved (P<0.01), differentiating myoblasts’ total protein (P<0.01) and CK activity (P<0.01) were markedly increased with actiin and daidzein concentration of 10ng/ml in medium, respectively. Meanwhile the expression of estrogen receptorβ(ER (3) in differentiating myoblasts’ was upgraded by the two phytoestrogens at the concentration of 10ng/ml in medium. These results suggest that arctiin increase skeletal muscle growth by increasing the proliferation of myoblasts and enhance protein synthesis and CK activity of differentiation myoblasts through binding to myocytes directly.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Chinese veterinary science > Veterinary Pharmacology
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