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Preparation method and application of inonotus obliquus polysaccharide

A technology of Inonotus obliquus and polysaccharide is applied in the field of preparation of Inonotus obliquus polysaccharide, which can solve problems such as unclear action mechanism, and achieve the effects of excellent antioxidant capacity, inhibition of aging and apoptosis, and broad application prospects.

Active Publication Date: 2019-05-14
JINLIN MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Inonotus obliquus has great development potential and application prospects in the fields of medicine and health products, but it is not clear which active component and its mechanism of action are specifically attributed to its wide range of pharmacological effects; therefore, providing a The preparation method of Inonotus obliquus polysaccharide, and studying its function is a problem urgently needed by those skilled in the art

Method used

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  • Preparation method and application of inonotus obliquus polysaccharide
  • Preparation method and application of inonotus obliquus polysaccharide
  • Preparation method and application of inonotus obliquus polysaccharide

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Experimental program
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Embodiment 1

[0036] The preparation of embodiment 1 Inonotus obliquus polysaccharide

[0037] (1) The response surface method was used to evaluate the combined effect of the polysaccharide extraction parameters of Inonotus obliquus based on single factor experiments. The parameter variables included extraction temperature, extraction time and liquid-solid ratio, and the polysaccharide yield was used as the response value to obtain the optimal extraction conditions It is: extraction time 1.5~2.5h, extraction temperature 100°C, liquid-solid ratio 30mL / g.

[0038](2) Hot water extraction of Inonotus obliquus polysaccharides according to the best extraction conditions obtained by response surface experiments: crush and sieve the dried Inonotus obliquus, and after degreasing with ethanol reflux, the residue is extracted in hot water at 100°C, and the liquid material The ratio is 30mL / g, the extraction time is 1.5~2.5h, the extract is filtered, the filtrate is combined, the filtrate is concentra...

Embodiment 2

[0039] Example 2 Establishing an in vitro antioxidant model to screen different polysaccharide active components of Inonotus obliquus

[0040] (1) Determination of DPPH free radical scavenging ability: 100 μL of 25-1600 μg / mL polysaccharide solution and vitamin E were added to a 96-well plate, and then 2×10 -4 mol L -1 100 μL of DPPH solution (prepared with absolute ethanol), mix well, and stand at room temperature in the dark for 30 minutes, measure the absorbance at a wavelength of 517 nm, and calculate the clearance rate according to the following formula:

[0041]

[0042] In the formula, A 3 Absorbance measured for different concentrations of polysaccharide plus DPPH, A 2 Absorbance measured with water for different concentrations of polysaccharides, A 1 Absorbance value measured for water plus DPPH, A 0 is the absorbance measured for water; the result is as Figure 4 As shown, the results showed that the DPPH free radical scavenging ability of IOP-1 was superior ...

Embodiment 3

[0046] Example 3 MTT cell viability assay

[0047] Rat aortic smooth muscle cells A10 (4,000 cells / well) in the logarithmic growth phase were seeded in a 96-well plate at 37°C with 5% CO 2 After culturing for 12-24 hours in an incubator, the cells were pretreated with vitamin E (Vitamin E) and 25-200 μg / mL polysaccharide IOP-1 of Inonotus obliquus at different concentrations, and a blank group was set up (only culture medium was added, no cells and IOP-1), control group (adding cells and culture medium, not adding IOP-1) and Eto model group (adding cells and culture medium), each group was provided with 5 duplicate wells. 24h later, 6μM Eto (etoposide, Etoposide is a cell cycle-specific antitumor drug that hinders DNA repair) was added to vitamin E, IOP-1 pretreatment group and Eto model group. It functions in the G1 phase and can be used as a model of senescence induction). After induction for 24 hours, incubate the cells with 0.5 mg / mL MTT at 37°C for 4 hours, remove MTT, ...

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Abstract

The invention discloses a preparation method and application of inonotus obliquus polysaccharide. Through water extraction and alcohol precipitation, repeated freeze thawing, deproteinization, dialysis, freeze drying, affinity chromatography and other methods, a polysaccharide component with antioxidant and anti-aging functions is separated and purified from inonotus obliquus; the inonotus obliquus polysaccharide component has strong antioxidant activity; through pretreatment of the inonotus obliquus polysaccharide, etoposide-induced proliferation inhibition and apoptosis of rat aortic smoothmuscle cells can be inhibited, and etoposide-induced cellular senescence beta-galactosidase activity increase, senescence-related heterochromatin aggregation and mitochondrial membrane potential decline can be inhibited; through pretreatment of the inonotus obliquus polysaccharide, etoposide-induced increase in expression levels of p53, p21 and p16 protein and mRNA can be inhibited. The inonotus obliquus polysaccharide components prepared according to the preparation method provided by the invention has good antioxidant and anti-aging activity and has a broad application prospect as a novel anti-aging protective medicine.

Description

technical field [0001] The invention relates to the field of natural product active ingredients, and more specifically relates to a preparation method and application of Inonotus obliquus polysaccharide. Background technique [0002] Inonotus obliquus (Fr.) Pilt is a rare and valuable medicinal fungus. , P. brown, P. obliquus, P. obliquus, Rosa, and Ganoderma lucidum, in the biological category, belong to the fungal phylum, basidiomycotina, laminomycetes, nonbryomycetes, polyporaceae , Brown Poria genus, mostly parasitic under the bark of birch, silver birch, elm, alder, etc. or on the dry after felling, mainly distributed in the area of ​​​​45°-50° north latitude. [0003] As early as the 12th century, the Manchu people used Inonotus obliquus to treat diseases. It is said that the Manchu people relied on Inonotus obliquus to treat various diseases when the mountains were closed by heavy snow. Over the years, Inonotus obliquus has been used in many countries to resist disea...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/715A61P39/06
Inventor 孙新任婷
Owner JINLIN MEDICAL COLLEGE
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