Application of eucommia ulmoides lignan extract in preparing PPARalpha agonist
A technology of high triglyceride and Eucommia lignan, which is applied in the field of medicine
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Embodiment 1
[0037] The preparation of embodiment 1 eucommia lignan extract
[0038] Take 2Kg of bark of Eucommia ulmoides (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 (column volume of macroporous resin is 3 L), wash with water for 3 times of column volume, continue to wash with 1% ammonia water for 3 times of 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 control, the lignan content of Eucommia ulmoides lignan extract was 85% by ultraviolet detection, and the content of pinoresinol diglucoside by HPLC was 12.5%.
Embodiment 2
[0039] The preparation of embodiment 2 eucommia lignan extract
[0040] After the bark of Eucommia ulmoides was crushed, 9 L of 60% ethanol was added to reflux for extraction for 1 h, filtered, and the filter residue was extracted with 7 L of 60% ethanol for 1 h, filtered, the two extracts were combined, and concentrated until there was no alcohol smell to obtain a concentrated solution of about 3 L. The concentrated solution was passed through the AB-8 macroporous adsorption resin column, firstly eluted with 5L of water, then 7L of 60% ethanol, collected the 60% ethanol eluate, concentrated to 150ml, slowly added to 450ml of water, filtered, concentrated, A dry solid powder was obtained. 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 the reduction of triglycerides by eucommia lignan extracts
[0044] 1.1 Experimental method
[0045] 1.1.1 Cell culture and grouping:
[0046] a) Culture HepG2 cells (37°C, 5% CO2) with DMEM high-glucose medium (containing 10% fetal bovine serum);
[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-sugar medium (containing 0.5 mM oleic acid, 10% fetal bovine serum) was added for cultivation, and the normal group was cultivated with DMEM low-sugar medium (containing 10% fetal bovine serum). Continue culturing for 24 hours to ...
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