阅读说明：本技术 一种抑制淀粉样蛋白产生的稀有人参皂苷及其应用 (Rare ginsenoside for inhibiting amyloid protein production and application thereof ) 是由 闫培生 陈琪琪 王婧莹 高秀君 周莹 曹立新 于 2021-08-26 设计创作，主要内容包括：本发明公开了一种抑制淀粉样蛋白产生的稀有人参皂苷的应用,稀有人参皂苷Compound K在益智、预防智力衰退、防治神经退行疾病等的功能食品、保健品和生物医药制品或其作为成分存在于组合物等中的应用。本发明发现稀有人参皂苷Compound K对Aβ42淀粉样蛋白形成的抑制效果好于β分泌酶抑制剂,从而可避免Aβ42淀粉样蛋白形成而影响智力。此外,稀有人参皂苷Compound K还具有一定的提高细胞活性的作用。(The invention discloses application of rare ginsenoside Compound K in the aspects of promoting intelligence, preventing intelligence deterioration, preventing and treating neurodegenerative diseases and the like, or application of the rare ginsenoside Compound K in functional foods, health-care products, biological medicine products or compositions and the like as a component. The invention finds that the inhibiting effect of the rare ginsenoside Compound K on the formation of the amyloid A beta 42 is better than that of a beta secretase inhibitor, thereby avoiding the influence of the formation of the amyloid A beta 42 on intelligence. In addition, the rare ginsenoside Compound K also has certain function of improving the cell activity.)

1. The application of rare ginsenoside for inhibiting amyloid protein production is characterized in that: the rare ginsenoside Compound K is applied to intelligence development, intelligence deterioration prevention and neurodegenerative diseases prevention.

2. The use of rare ginsenosides for inhibiting amyloid production according to claim 1, wherein: the rare ginsenoside Compound K is applied to inhibiting the generation of A beta 42 amyloid.

3. The use of rare ginsenosides for inhibiting amyloid production according to claim 1, wherein: the rare ginsenoside Compound K is used for preparing a beta secretase or gamma secretase inhibitor and a composition.

4. Use of rare ginsenosides for inhibiting amyloid production according to any of claims 1-3, characterized in that: the rare ginsenoside Compound K is independently used in functional foods, health products and biological medicine products or the ginsenoside Compound K exists in the composition as a component.

Technical Field

The invention relates to a bioactive substance and application thereof, in particular to application of rare ginsenoside Compound K for inhibiting amyloid protein generation, which can be applied to the fields of functional foods, health-care products, biological medicine products and the like.

Background

The rare ginsenoside Compound K is a ginsenoside which does not exist or has a very small content in natural ginseng and American ginseng, and as the activity of the ginsenoside Compound K is researched more and more extensively, the rare ginsenoside Compound K shows better biological activity in many aspects, such as: can induce the apoptosis of tumor cells to play a good role in resisting tumors; acting on hyaluronic acid synthetase to promote hyaluronic acid secretion, thereby playing good roles of delaying skin dryness, resisting wrinkles and the like; and has antiallergic, antiinflammatory, and hepatoprotective effects.

There is no research on the related studies of rare ginsenoside Compound K in neurodegenerative diseases.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention provides the application of rare ginsenoside for inhibiting the generation of amyloid protein.

In order to solve the technical problems, the invention adopts the technical scheme that: application of rare ginsenoside Compound K in improving intelligence, preventing intelligence deterioration, and preventing and treating neurodegenerative diseases is provided.

Furthermore, the rare ginsenoside Compound K is independently used for preparing functional foods, health care products and biological medicine products which have the functions of inhibiting the generation of A beta 42 amyloid, benefiting intelligence, preventing intelligence decline, preventing neurodegenerative diseases and the like.

Further, the rare ginsenoside Compound K is used for preparing a composition of functional food, health care products and biological medicine products which can inhibit the generation of A beta 42 amyloid, promote intelligence, prevent intelligence deterioration, prevent neurodegenerative diseases and the like.

Further, the rare ginsenoside Compound K is used for preparing beta secretase or gamma secretase inhibitors and compositions.

The invention discovers that the rare ginsenoside Compound K has the function of inhibiting cells from generating amyloid A beta 42, and avoids the intelligence decline and the attack of neurodegenerative diseases caused by the formation of the amyloid A beta 42. In addition, the rare ginsenoside Compound K also has the function of improving the cell activity.

Drawings

FIG. 1 is a calibration curve prepared by measuring the OD450nm values at different concentrations of the amyloid A β 42 as a standard.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.

Application of rare ginsenoside for inhibiting amyloid protein production is provided.

The invention provides application of rare ginsenoside Compound K in intelligence development, intelligence deterioration prevention and neurodegenerative diseases prevention.

Comprises rare ginsenoside Compound K used for preparing functional food, health product and biological medicine product or composition thereof for inhibiting A beta 42 amyloid protein generation, improving intelligence, preventing intelligence deterioration, and preventing and treating neurodegenerative diseases. And rare ginsenoside Compound K used for preparing beta secretase or gamma secretase inhibitors and compositions.

Wherein, the functional food, health care product and biological medicine product or the composition thereof which is prepared from the rare ginsenoside Compound K and has the functions of inhibiting the generation of A beta 42 amyloid protein, promoting intelligence, preventing intelligence deterioration, preventing and treating neurodegenerative diseases and the like can be in any acceptable dosage form, including oral dosage forms and external dosage forms. The oral dosage forms comprise tablets, tabletting candies, hard capsules, soft capsules, oral liquid, syrup, granules, dropping pills, orally disintegrating tablets, sustained release capsules, controlled release tablets, controlled release capsules and the like. The external preparation includes water injection, freeze dried powder for injection, aseptic powder for injection, transfusion, etc.

Effect of ginsenoside Compound K on cell viability

Preparing materials:

collecting cells in logarithmic growth phase, removing cell culture solution, digesting with 0.25% pancreatin at 37 deg.C for 2-3 min, adding 2mL DMEM culture solution to obtain single cell suspension, counting cells under microscope, and adding culture solutionAdjusting the cell concentration to 2.5X 10⁵Each well of 96-well cell culture plates was incubated with 200. mu.L of cell suspension at 37 ℃ for 6 hours in a carbon dioxide incubator containing 5% carbon dioxide, and then the culture medium was aspirated and washed 1 time with DMEM pre-warmed at 37 ℃.

Experimental treatment:

experimental groups: DMEM culture solutions containing rare ginsenoside compound K with effective concentrations of 5, 10 and 20 mug/mL are added to the cell material respectively.

Control group: the cell material is added with a culture medium.

The treatment method comprises the following steps:

and (3) processing each group of the experimental group and the control group for 3 multiple wells, continuously culturing for 12h, adding 20 mu L of MTT (methyl thiazolyl tetrazolium) with the concentration of 5mg/mL into each well, continuously culturing for 4h, then removing supernatant, adding 200 mu L of DMSO into each well, and oscillating on a microplate oscillator for 10min until crystals are completely dissolved.

The 96-well culture plate is placed on a microplate reader, the optical density value of each well is measured at 490nm, and the cell activity is calculated according to the optical density value of each well.

The formula for calculating the cell activity is: percent cell activity is defined as the optical density of the experimental samples/the optical density of the control x 100%.

The experimental results are as follows:

the cell activity was calculated from the optical density values of the wells measured by the microplate reader as shown in table 1, and it was found that the ginsenoside compound K had an effect of improving the cell activity.

TABLE 1 Effect of different concentrations of ginsenoside compound K on cell viability

Inhibition of A beta 42 amyloid formation by (II) rare ginsenoside Compound K

Experimental treatment:

experimental groups: taking cells in logarithmic growth phase, adding DMEM culture solution containing rare ginsenoside Compound K with effective concentration of 5, 10 and 20 mug/mL respectively.

Negative control group: DMEM medium was added to the cells in the logarithmic growth phase.

Positive control group: adding DMEM culture solution containing beta secretase inhibitor Verubechsat with the effective concentration of 20 mu g/mL into cells in the logarithmic growth phase; verubechsat is a potent and selective inhibitor of beta-secretase.

The treatment method comprises the following steps:

each of the above treatments was repeated three times in the above groups, and after all treatments were incubated for 12 hours at 37 ℃ in a carbon dioxide incubator with 5% carbon dioxide concentration, the supernatant was collected and used for the determination of the A.beta.42 content.

According to the operation requirement of a 'human beta amyloid polypeptide 42 (Abeta 42) Elisa kit', A beta 42 contents of a standard sample and a test sample are measured, a standard curve of the standard sample is prepared, the content (pg/ml) of the A beta 42 of the test sample is calculated according to the standard curve, and the inhibition rate of the test sample on the A beta 42 is calculated by the following formula:

the inhibition ratio [% ], [% ] is the content of the negative control a β 42-the content of the experimental group or the positive control a β 42/the content of the negative control a β 42 x 100%.

The experimental results are as follows:

table 2 OD values of different concentrations of standard a β 42 amyloid protein determined by the kit.

OD standard curves were plotted according to table 2 for different concentrations of standard a β 42 amyloid, as shown in fig. 1.

The standard calculation formula is obtained according to the standard curve in fig. 1:

Y＝0.0032x+0.1312,

R²＝0.9866；

the OD values measured by the "human amyloid-beta-polypeptide 42(a β 42) Elisa kit" for the experimental group and the positive control group were substituted into the formula to obtain the corresponding amyloid concentration of a β 42.

And calculating the inhibition rate of the samples of the experimental group and the positive control group on the generation of the amyloid A beta 42 by using an inhibition rate calculation formula on the generation of the amyloid A beta 42, wherein the inhibition rate is shown in Table 3:

table 3 inhibition of a β 42 amyloid production by positive control and experimental groups.

And (4) conclusion: the experimental results show that the rare ginsenoside Compound K has obvious inhibition effect on the formation of Abeta 42 amyloid, and the inhibition effect is superior to that of a beta secretase inhibitor (Verubechestat), so the ginsenoside Compound K has beneficial effects on the aspects of intelligence development, intelligence deterioration prevention, neurodegenerative disease prevention and the like, and can be applied to functional foods, health care products, biological medicine products or compositions thereof and the like for intelligence development, intelligence deterioration prevention, neurodegenerative disease prevention and the like.

The above embodiments are not intended to limit the present invention, and the present invention is not limited to the above examples, and those skilled in the art may make variations, modifications, additions or substitutions within the technical scope of the present invention.