Preparation and application of beta-conglycinin-procyanidine B2 compound
A technology of conglycinin and proanthocyanidin, which is applied in the field of preparation of β-conglycinin-proanthocyanidin B2 complex, can solve the problems of toxic side effects and limited application, and achieve the effects of easy production and preparation, broad prospects and wide sources
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Embodiment 1
[0029] Example 1 Preparation of β-conglycinin-procyanidin B2 complex
[0030] 1. Preparation of β-conglycinin
[0031] 1. Take Dongnong No. 47 soybeans, peel them with a peeling machine, and then pulverize them into powder with a pulverizer, pass through a 60-mesh sieve, and then mix the soybean powder with acetone at 1:5 (g / mL) and let stand for reaction for 1 hour, and filter to obtain degreasing. soy flour;
[0032] 2. Mix defatted soybean powder and water at 1:15 (g / mL), adjust the pH to 7.5, stir on a magnetic stirrer for 1 h, centrifuge (9000 rpm, 20 min, 4 °C), absorb the supernatant and add NaHSO 3 (0.98g / L) adjust pH to 6.4 and place at 4℃ overnight;
[0033] 3. Continue centrifugation on the second day (6500rpm, 20min, 4℃), then absorb the supernatant and add NaCl (0.25×58.44×volume) to adjust the pH to 5.0, stir on a magnetic stirrer for 1h, and continue centrifugation (9000rpm, 30min, 4 °C);
[0034] 4. Aspirate the supernatant and add 2 times deionized water t...
Embodiment 2
[0037] Example 2 Establishment of high-fat model of HepG2 cells
[0038] 1. The principle of establishing the high-fat model of HepG2 cells
[0039]At the cellular level, HLP manifested as increased intracellular TG and TC content, which resulted in the inability of cells to achieve normal lipid metabolism. In the process of body metabolism, the liver is the main site for the synthesis of TG and TC in the human body. HepG2 cells are a cell line composed of human hepatoma cells, which express 3-hydroxy-3-methylglutarate coenzyme A reductase. Statins achieve the effect of lowering blood lipids by inhibiting the activity of reductase. In addition, HepG2 cells have more lipid content on the mesangium than other cell membranes, because HepG2 cells express apoA-1, which is the main component of high-density lipoprotein. It is a kind of apolipoprotein, which is a kind of cholesterol that is beneficial to the body. It transports cholesterol and lipids in cells to the liver for decomp...
Embodiment 3
[0050] Example 3 Beta-conglycinin-procyanidin B2 complex assisted hypolipidemic function test
[0051] 1. The effect of β-conglycinin and procyanidin B2 on the survival rate of HepG2 cells
[0052] Cells were digested with 0.25% trypsin, counted, and HepG2 was added to 100 μL per well at a cell density of 5×10 4 cells / mL were inoculated into 96-well plates (one plate each for β-conglycinin and procyanidin B2), cultured for 24 h, and after the cells adhered to the wall, the culture medium was discarded;
[0053] Set up experimental group, model group (MC) and control group (BC):
[0054] 1) The experimental group was added with different concentrations of β-conglycinin and procyanidin B2 prepared with DMEM medium containing 1% FBS;
[0055] 2) The model group was supplemented with oleic acid modeling agent with a final concentration of 1000 μmol / L prepared with DMEM medium containing 1% FBS;
[0056] 3) The control group is DMEM medium containing 1% FBS;
[0057] Each group...
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