Application of eucommia ulmoides lignan extract in preparing PPARalpha agonist
A technology of Eucommia lignan and extract, applied in the field of medicine
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Embodiment 1
[0037] Example 1 Preparation of Eucommia Lignan Extract
[0038] Take 2Kg of eucommia bark (with the outer skin removed), extract twice with 8L of 65% ethanol under reflux, each time for 1h, filter, combine the two extracts and concentrate to 800ml. Mix and load the sample with D101 macroporous resin (macroporous resin column volume 3L), wash with water for 3 times the column volume, continue to wash with 1% ammonia water for 3 times the column volume, and then wash with water until neutral. Elute with 20% ethanol for 2 column volumes and 45% ethanol for 2.5 column volumes. The 45% ethanol eluate was concentrated to 600ml. The eluate was concentrated and dried to a powder. Using pinoresinol diglucoside as a contrast, the lignan content of Eucommia ulmoides lignan extract was 85% detected by ultraviolet light, and the content of pinoresinol diglucoside detected by HPLC was 12.5%.
Embodiment 2
[0039] Example 2 Preparation of Eucommia Lignan Extract
[0040]After the bark of Eucommia ulmoides is crushed, add 9L of 60% ethanol to reflux for extraction for 1 hour, filter, add 7L of 60% ethanol to the filter residue, extract for 1 hour, filter, combine the two extracts, concentrate until there is no alcohol smell, and obtain a concentrated solution of about 3L. The concentrated solution was passed through an AB-8 macroporous adsorption resin column, firstly eluted with 5L of water, then 7L of 60% ethanol, collected the 60% ethanol eluate, concentrated to 150 ml, slowly added to 450 ml of water, filtered, Concentrate to obtain dry solid powder. Using pinoresinol diglucoside as a control, the lignan content of Eucommia ulmoides lignan extract was 51% detected by ultraviolet light, and the content of pinoresinol diglucoside detected by HPLC was 3.5%.
Embodiment 3
[0042] The eucommia lignan extracts obtained in Example 1 and Example 2 were used in the experimental research of lowering triglyceride and the experimental research of activating PPARα in Example 3.
[0043] 1. Experimental Study on Lowering Triglyceride of Eucommia Lignan Extract
[0044] 1.1 Experimental method
[0045] 1.1.1 Cell culture and grouping:
[0046] a) Culture HepG 2 cells with DMEM high glucose medium (containing 10% fetal bovine serum) (37 ℃, 5% CO 2 );
[0047] b) HepG 2 was inoculated in a 6-well plate at a certain density of 5000 / mL, 1 ml / well, and cultured with DMEM high-glucose medium (containing 10% fetal bovine serum);
[0048] c) When the cells were inoculated for 24 hours, the old medium was discarded, and DMEM low-glucose medium (containing 0.5 mM oleic acid, 10% fetal bovine serum) was added for culture, and the normal group was cultured with DMEM low-glucose medium (containing 10% fetal bovine serum) The culture was continued for 24 h for mode...
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