Application of radix acanthopanacis trifoliate extract in inhibiting alpha-glucosidase
A technology of glucosidase and extract, which is applied in the field of application of bok choy extract to inhibit α-glucosidase, and can solve problems such as no blood sugar lowering effect of bok choy
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Embodiment 1
[0029] Preparation of bok choy extract
[0030] Collect fresh leaves, stems, and roots of bougainvillea, dry them at room temperature, crush them, take out 200g each, add 2L 95% edible alcohol for cold extraction, filter every other day, extract three times in total, combine the extracts, and concentrate them with a rotary evaporator The ethanol extracts of different parts (leaf: L; stem: S; root: R) were obtained. Each ethanol extract was sequentially extracted with petroleum ether, ethyl acetate and n-butanol, concentrated and dried to obtain petroleum ether layer (leaf: LP; stem: SP; root: RP), ethyl acetate layer (leaf: LE; stem: RP). : SE; root: RE), n-butanol layer (leaf: LN; stem: SN; root: RN). All extracts were vacuum-dried and stored at 4°C for future use.
Embodiment 2
[0032] Chemical Constituents of Bougainvillea Extract
[0033] The extract of the ethyl acetate layer of bok choy was separated and purified by repeated silica gel column chromatography, Sephadex LH-20 column chromatography, reversed phase silica gel column chromatography, preparative thin layer chromatography (pTLC), MCI resin column chromatography and other separation methods. To obtain monomer compounds, comprehensively use modern spectrum techniques (IR, UV, MS, 1 H-NMR, 13 C-NMR, DEPT, 1 H- 1 H COZY, HSQC, HMBC) and compared with the literature, the structures of 9 known terpenoids were determined, and their chemical names were impressive acid (1); 3α,11α-dihydroxy-lup-20(29)- en-23-al-28-oic acid(2); 3α-hydroxy-lup-20(29)-en-23,28-dioic acid(3); 3α,11α-dihydroxy-lup-20(29)- en-23,28-dioic acid(4); 3α-hydroxy-lup-20(29)-en-30-ol-23,28-dioic acid(5); ent-kaur-15-en-17-al -19-oic acid (6); 17-hydroxy-16α-ent-kauran-19-oic acid (7); 16α-hydroxy-ent-kauran-19-oic acid (8...
Embodiment 3
[0036] Inhibitory activity of bok choy extract on α-glucosidase
[0037]Preparation of α-glucosidase solution: Dissolve the lyophilized enzyme powder in 0.1% BSA solution to prepare 100 U / mL enzyme solution, and store it in a refrigerator at -20°C. Before the experiment, pipette a small amount of 100U / mL enzyme solution, first dilute to 20U / mL with 0.1% BSA solution, and then dilute to 0.2U / mL with 0.1% BSA solution for use.
[0038] Preparation of PNPG solution: Weigh a certain amount of PNPG solid, dissolve it with 0.1mol / L pH 6.8 phosphate buffer saline (PBS) to make a concentration of 10mmol / L, and then dispense it into 1.5mL centrifuge tubes.
[0039] In this experiment, the activity of α-glucosidase was determined by colorimetric method. The bok choy extract or compound to be tested was dissolved in dimethyl sulfoxide (DMSO) to prepare a stock solution with a mass concentration of 10 mg / mL. Then dilute the stock solution with PBS to prepare the solution to be tested at...
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