Polygala arillata saponin E, extract containing same and application
A technology for extracts and root extracts, applied in the field of new compounds, can solve the problems of large toxic and side effects, limited drug efficacy, and small toxic and side effects
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Embodiment 1
[0040] Example 1 Extraction and Characterization of Compound Polygala Saponin E
[0041] 1. Instruments and materials
[0042] X-5 Microscopic Melting Point Meter (Beijing Tektronix Instrument Company); ZF-20D Dark Box UV Analyzer (Gucun Electro-optical Instrument Factory); BrukerAV-III NMR Spectrometer, TMS as internal standard (Swiss Bruker Company); Synapt G2Mass mass spectrometer (U.S. Waters company); Agilent1260 high performance liquid chromatograph (U.S. Agilent company); SunfireTMPrep C18 chromatographic column (19 * 150mm, 5 μ m); Waters2489 preparative liquid chromatography (U.S. Waters company); Column chromatography uses silica gel (Qingdao Ocean Chemical Factory); silica gel GF254 thin-layer prefabricated plate (Yucheng Chemical Shanghai Co., Ltd.); Sephadex LH-20 (Beijing Suo Laibao Technology Co., Ltd.); the reagents used are analytically pure and chromatographically pure.
[0043] The Osmanthus fragrans of Hebao Mountain is collected from Yunnan Province, and ...
Embodiment 2
[0058] Example 2 Anti-inflammatory activity test of polygala saponin E
[0059] 1. Experimental materials
[0060] Polygala saponin E pure product of Polygala saponin E prepared by the extraction method in Example 1; Polygala saponin E stock solution: Dissolve Polygala saponin E in DMSO, and prepare a 1 mg / ml solution or a uniform solution with PBS buffer Mixed solution; mouse macrophage RAW 264.7 (Binhai Laboratory, Tianjin University of Traditional Chinese Medicine); culture medium: DMEM high glucose medium (U.S. Hyclone Company)+10% fetal bovine serum (U.S. Hyclone Company)+1% double antibody ( HyClone, USA); FlexStation 3 automatic microplate reader (Molecular Devices, USA); Griess Reagent System kit (Promega, USA); bacterial lipopolysaccharide (LPS, Sigema, USA, catalog number: L2880).
[0061] 2. Experimental method
[0062] NO is involved in many physiological and pathological processes. Macrophages can produce a large amount of NO under the induction of LPS. At the s...
Embodiment 3
[0071] Example 3 Cytotoxic activity test of polygala saponin E
[0072] 1. Experimental materials
[0073] Cell Counting Kit-8 kit (Dojindo Company, Japan).
[0074] 2. Experimental method
[0075] 1) Take the cells in the logarithmic growth phase and gently scrape them off with a cell scraper, and inoculate them evenly in a 96-well plate (no cells are inoculated on the edge, and filled with sterile PBS), 100 μL per well, and the number of inoculated cells is about 1×10 4 pc / well 37°C, 5% CO 2 Cultivate in the incubator for 2 hours;
[0076] 2) Divide the cells into the polygala saponin E group, the DXM group, the control group (control) and the LPS group; after the cells adhere to the wall, suck out the culture medium in the well; then add the polygala saponin E group containing different final concentrations of The culture solution of polygala saponin E (the final concentration of polygala saponin E was 50 μg / mL, 25 μg / mL and 12.5 μg / mL); the DXM group was added with the...
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