阅读说明：本技术 刺五加提取物在制备神经退行性相关疾病药物的应用 (Application of acanthopanax senticosus extract in preparation of medicines for treating neurodegenerative related diseases ) 是由 周慧 付佳 何洋 张欣 于 2020-09-01 设计创作，主要内容包括：本发明属于医药用途及制备技术领域,具体涉及刺五加提取物在制备神经退行性相关疾病药物的应用。本发明通过细胞和动物试验发现：刺五加水提取物,经大孔树脂30％乙醇洗脱组分具有协同作用,作为治疗帕金森疾病的刺五加的最佳有效部位,对于神经退行性相关疾病药物的应用具有优异的效果。(The invention belongs to the technical field of medical application and preparation, and particularly relates to application of an acanthopanax senticosus extract in preparation of medicines for treating neurodegenerative related diseases. The invention discovers that: the acanthopanax water extract has a synergistic effect by eluting the components with macroporous resin and 30% ethanol, is used as the optimal effective part of the acanthopanax for treating the Parkinson's disease, and has an excellent effect on the application of medicines for treating neurodegenerative related diseases.)

1. The application of the acanthopanax senticosus extract in preparing medicines for treating neurodegenerative related diseases is characterized in that the acanthopanax senticosus extract is obtained by the following preparation method: extracting dry radix Acanthopanacis Senticosi with water under reflux, filtering to obtain supernatant, precipitating with 70% ethanol overnight, concentrating the supernatant, separating with macroporous resin column, eluting with 10% and 30% ethanol water solution respectively to obtain 30% ethanol eluate, and lyophilizing.

2. The use according to claim 1, wherein the neurodegenerative-related diseases include, but are not limited to, alzheimer's disease, parkinson's disease, amyotrophic lateral sclerosis, huntington's disease.

3. Use according to claim 1, wherein the macroporous resin is preferably an AB-8 macroporous resin.

4. Use according to claim 1, wherein the amount of eluent is 5 times the volume of the resin column.

5. Use according to claim 1, wherein the elution flow rate is 4 BV/h.

6. The use of claim 1, wherein the weight ratio of the acanthopanax senticosus to the water in the heating reflux extraction process is 3: 10.

7. the use according to claim 1, wherein the water is soaked for more than 0.5h before the reflux extraction.

8. The use of claim 1, wherein the reflux extraction process is performed under slightly boiling conditions for 1 hr, and the supernatant is obtained by filtration.

9. The use of claim 1, wherein the residue is subjected to first heating reflux extraction, then distilled water is added for the first extraction, the supernatant is obtained by extraction and filtration in the same way, the obtained medicinal liquids are combined and concentrated for 2 times to obtain the total water extract of radix Acanthopanacis Senticosi, and then the total water extract is precipitated by ethanol with concentration of 70% or more.

10. The use as claimed in claim 1, wherein the effective component of Acanthopanax senticosus harms extract contains flavone 30.91% ± 0.76% calculated as rutin, and saponin 28.97% ± 2.6% calculated as oleanolic acid.

Technical Field

The invention belongs to the technical field of medical application and preparation, and particularly relates to application of an acanthopanax senticosus extract in preparation of medicines for treating neurodegenerative related diseases.

Background

At present, with the progress of global aging accelerating, age-related neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, etc.) have become serious diseases that seriously endanger the health of human life and reduce the quality of life. Among them, there are over 400 million patients with Parkinson's disease worldwide, and 170 million of them. Research shows that the number of people suffering from the disease in big cities in China is increased dramatically in recent 20 years. It is estimated that nearly 10 million people in China become new Parkinson's disease patients every year. After the patient suffers from tremor, stiffness, slow movement, dyskinesia and vegetative nerve symptoms, the disease condition is progressively aggravated, the mobility of the patient is seriously limited, the life quality of the patient is influenced, great pain is caused to the patient, and serious burden is brought to the society and families, so that an effective traditional Chinese medicine for preventing and treating the Parkinson disease is urgently needed to delay the progress of the Parkinson disease and prevent the aggravation of the disease condition.

Dried roots of Acanthopanax senticosus (Rupr. Et Maxim.) Harms. Mainly produced in mountain areas of the province of Heilongjiang. Has pungent, slightly bitter, warm and nontoxic properties, and has effects of invigorating kidney, invigorating spleen, invigorating kidney and tranquilizing mind. At present, the acanthopanax compound and the acanthopanax injection are used for the clinical treatment of the Parkinson disease and have certain curative effect.

Disclosure of Invention

In view of the problems in the prior art, the invention provides the application of the acanthopanax senticosus extract in preparing medicines for treating neurodegenerative related diseases, wherein the acanthopanax senticosus extract is prepared by the following preparation method: extracting dry radix Acanthopanacis Senticosi with water under reflux, filtering to obtain supernatant, precipitating with 70% ethanol overnight, concentrating the supernatant, separating with macroporous resin column, eluting with 10% and 30% ethanol water solution respectively to obtain 30% ethanol eluate, and lyophilizing.

As a preferred embodiment of the present invention, the neurodegenerative-related diseases include, but are not limited to, alzheimer's disease, parkinson's disease, amyotrophic lateral sclerosis, huntington's disease, preferably parkinson's disease.

As a preferred embodiment of the present invention, the macroporous resin is preferably AB-8 macroporous resin.

In a preferred embodiment of the present invention, the amount of eluent is 5 times the volume of the resin column.

As a preferred embodiment of the present invention, the elution flow rate is 4 BV/h.

As a preferred embodiment of the invention, the mass ratio of the acanthopanax senticosus to the water in the heating reflux extraction process is 3: 10.

in a preferred embodiment of the present invention, the extract is soaked in water for 0.5 hours or more before the reflux-heating extraction.

As a preferred embodiment of the invention, the heating reflux extraction process keeps the reflux for 1 hour in a slightly boiling state, and the supernatant is obtained by filtration.

As a preferred embodiment of the present invention, the filtered residue after the first heating reflux extraction is added with the amount of distilled water used for the first extraction, and the same method is used for extraction and filtration to obtain the supernatant, and the obtained liquid medicines obtained 2 times are combined and concentrated to obtain the total water extract (i.e. stock solution) of acanthopanax root, and then the total water extract is precipitated by ethanol with the concentration of more than 70%.

As a preferred embodiment of the invention, the content of flavone in the effective component of the acanthopanax senticosus extract is 30.91% + -0.76% in terms of rutin, and the content of saponin is 28.97% + -2.6% in terms of oleanolic acid.

The beneficial effects of the invention compared with the prior art comprise:

the invention unexpectedly finds that 30% of the components are used as the best effective parts of acanthopanax for treating the Parkinson's disease, and researches on chemical components of the 30% of the components find that the flavone and saponin components with the compositions are not the highest among the separation by various methods, but unexpectedly show the best treatment effect, thus the excellent treatment effect is shown through synergistic action due to the existence of various substances in the product.

The invention passes spontaneous activity experiments, pole climbing experiments, rolling shaft experiments and suspension experiments. The mice in the model group have obvious behavioral disturbance, the coordination movement capability is reduced, the times of autonomous activities and the times of spontaneous standing are reduced, the climbing, rolling and hanging time is prolonged, and the result shows that 30 percent of the components of the acanthopanax root have obvious anti-Parkinson effect. The results of cell experiments prove the prevention and treatment effects of acanthopanax on Parkinson.

Drawings

FIG. 1 shows a standard curve of rutin in a control sample;

FIG. 2, standard curve of oleanolic acid as a control;

FIG. 3, MPP⁺Effect of different concentrations on cell viability

FIG. 4 shows the effect of the effective components of Acanthopanax senticosus root on oxidative stress injury model

FIG. 5 shows the effect of effective components of Acanthopanax senticosus root on anoxia-sugar deficiency model

FIG. 6, Effect of 30% fraction on the number of spontaneous standings in Parkinson's mice

FIG. 7, effect of 30% fraction on the number of spontaneous Activities in Parkinson's mice

FIG. 8, effect of 30% fraction on Pole-climbing time in Parkinson's mice

FIG. 9, effect of 30% fraction on suspension time in Parkinson's mice

FIG. 10, effect of 30% composition on Rolling shaft time in Parkinson's mice

Wherein, in each figure, p<0.01，^#p<0.05, to model set.

Detailed Description

The present invention is described in further detail with reference to the following specific examples and accompanying drawings, but the embodiments of the invention are not limited by the examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and they are included in the scope of the present invention.

Laboratory instruments and materials

Nitrogen blowing apparatus (HSC-24A), AB-8 macroporous adsorbent resin (Shandong Lukang stereo pharmaceutical Co., Ltd.), Spectra FLOUR plus enzyme-linked immunosorbent assay (TECAN, Switzerland), GENESYS 10S visible spectrophotometer (Shanghai Jun-wing instruments Co., Ltd.), SanoriusBSl10S analytical balance (Beijing Sidoristi Co., Ltd.), XZR-A rotary evaporator (Corrony instruments Co., Ltd.), ALPHA 2-4 freeze dryer (Marin Christ Co., Ltd.), KDM type electric mantle (Juglau Hualu electric heating instruments Co., Ltd.), 96 well plate (thermo Co., Ltd.)

Rutin (201004 Vickqi Biotech Co., Sichuan province), oleanolic acid (200304, institute for testing biological products of Chinese medicine), perchloric acid, glacial acetic acid, and anhydrous sodium chlorideEthanol, sodium nitrite, aluminum nitrate, sodium hydroxide, ammonium bicarbonate, ferrous sulfate, methanol, glacial acetic acid, hydrochloric acid, ferric chloride, disodium hydrogen phosphate, ferric chloride and sodium dihydrogen phosphate are analytically pure and purchased from Beijing chemical plant, 30% hydrogen peroxide solution (Beijing chemical plant), 1, 1-diphenyl-2-picrylhydrazino (DPPH) and disodium ethylene diamine tetraacetate (EDTA-Na)₂) (Sigma Co.), pyrogallol (Aladdin reagent Co., Ltd.), WST-1 (Nanjing optical technology Co., Ltd.), ascorbic acid (VC), basic saffron (saffron T) (Beijing Dingguo Changsheng biotechnology Co., Ltd.), sodium acetate (Yili fine chemicals Co., Ltd.), 4, 6-tripyridyltriazine (TPTZ) (Shanghai Jing pure practice Co., Ltd.).

Dried root of Acanthopanax senticosus (Rupr. Et Maxim.) Harms was purchased from Beijing Hojingtang drugstore and identified by professor Wang Shumin, university of traditional Chinese medicine, Changchun.