CN117487035A - Ginseng polysaccharide and preparation method and application thereof - Google Patents
Ginseng polysaccharide and preparation method and application thereof Download PDFInfo
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- CN117487035A CN117487035A CN202311443594.9A CN202311443594A CN117487035A CN 117487035 A CN117487035 A CN 117487035A CN 202311443594 A CN202311443594 A CN 202311443594A CN 117487035 A CN117487035 A CN 117487035A
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Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/08—Antiepileptics; Anticonvulsants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/24—Antidepressants
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Epidemiology (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Pain & Pain Management (AREA)
- Materials Engineering (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
- Psychiatry (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Sustainable Development (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides ginseng polysaccharide, a preparation method and application thereof, and relates to the technical field of medicines. The preparation method of the ginseng polysaccharide comprises the following steps: pulverizing Ginseng radix, reflux-extracting with hot water, concentrating, centrifuging, precipitating supernatant with ethanol solution, dialyzing the precipitate with distilled water, and lyophilizing non-dialysate to obtain ginseng polysaccharide; the molecular weight cut-off of the dialysis was 3.5kDa. The ginseng polysaccharide prepared by the invention has the functions of regulating intestinal flora, resisting depression and resisting epilepsy. The experimental result of the ginseng polysaccharide shows that the ginseng polysaccharide prepared by the invention can be used as a potential medicament for treating depression and epilepsy.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to ginseng polysaccharide, and a preparation method and application thereof.
Background
Depression is a syndrome characterized by a sustained fall in mood, and is characterized by symptoms such as reduced interest, lassitude, anxiety, self-responsibility, and the like. Depression has become a very high incidence of disease due to the tension and strong competitive pressure of modern life rhythms, estimated by the World Health Organization (WHO), 5% of all adults worldwide suffer from depression. The disease burden in mental disorders is the leading one in global depressive disorder; depressive disorders rank 13 in the disease burden of all diseases worldwide.
Anxiety is one of the most common co-morbidities of epilepsy. The prevalence of anxiety associated with adult epileptic patients is about 11.0% to 39.4%. The bi-directional association between epilepsy and depression suggests that both may have a common neurobiological basis and that there may be some co-morbid mechanism. There are studies that abnormal frontal and temporal lobe structures and reduced secretion of neurotransmitters (gamma aminobutyric acid, 5-hydroxytryptamine, norepinephrine and dopamine) in the brain are considered to be involved in the occurrence of epilepsy and anxiety depression.
The microflora in the human intestinal tract is huge in number and complex in structure and is called the second brain of human beings. More and more studies have shown that intestinal microorganisms can influence brain function and behaviour through the flora-gut-brain axis. Interactions between gut microorganisms, neurons and behavioral changes may help to ameliorate social deficits caused by anxiety, depression, autism, etc. In another study published by Mazmanian laboratories, nature, 2021, a specific set of neuronal circuits has been identified that are directly affected by intestinal flora and subsequently lead to anti-social behaviour in mice lacking a specific intestinal flora.
The Ginseng Polysaccharide (GP) is one of important bioactive components of ginseng of Araliaceae, and is a high molecular dextran; has various pharmacological effects of resisting tumor, regulating immunity, resisting radiation, reducing blood sugar, etc., and has the effects of resisting adhesion, resisting virus, resisting oxidation, reducing inflammatory reaction, resisting septicemia, etc., and is clinically used for the comprehensive treatment of various malignant tumors, and relieving adverse reactions caused by chemotherapy and radiotherapy. Panaxan has been reported to modulate intestinal microbiota and make αpd-1 mab immunotherapy more sensitive to treatment in non-small lung cancer cells. However, research on regulation of intestinal flora, depression and epilepsy by ginseng polysaccharides has not been reported yet.
Disclosure of Invention
Therefore, the invention aims to provide ginseng polysaccharide, and a preparation method and application thereof, wherein the prepared ginseng polysaccharide has the effects of regulating intestinal flora, resisting depression and resisting epilepsy.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of ginseng polysaccharide, which comprises the following steps: pulverizing Ginseng radix, reflux extracting with hot water, concentrating, centrifuging, precipitating supernatant with ethanol solution, dialyzing the precipitate with distilled water, and lyophilizing non-dialysate to obtain ginseng polysaccharide; the molecular weight cut-off of the dialysis was 3.5kDa.
Preferably, the ratio of the feed liquid of the hot water reflux extraction is 8-10 g/mL, the temperature is 100 ℃, the time is 2 hours, and the times are 1-3 times.
Preferably, the rotation speed of the centrifugation is 2500-3500 rpm, and the time is 20-40 min.
Preferably, the volume percentage of the ethanol solution is 75-85%.
Preferably, the volume ratio of the supernatant to ethanol solution extraction is 1:4.
The invention also provides ginseng polysaccharide obtained by the preparation method.
Preferably, the molecular weight of the ginseng polysaccharide is 627kDa.
The invention also provides application of the ginseng polysaccharide in preparing a product for regulating intestinal flora.
The invention also provides application of the ginseng polysaccharide in preparation of antidepressant drugs.
The invention also provides application of the ginseng polysaccharide in preparing medicines for treating epilepsy.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a preparation method of ginseng polysaccharide, which comprises the following steps: pulverizing Ginseng radix, reflux-extracting with hot water, concentrating, centrifuging, extracting supernatant with ethanol solution, dialyzing the precipitate with distilled water, and lyophilizing non-dialysate to obtain ginseng polysaccharide; the molecular weight cut-off of the dialysis was 3.5kDa. The ginseng polysaccharide prepared by the invention has the functions of regulating intestinal flora, resisting depression and resisting epilepsy. The experimental result of the ginseng polysaccharide shows that the ginseng polysaccharide prepared by the invention can be used as a potential medicament for treating depression and epilepsy.
Drawings
FIG. 1 is a High Performance Liquid Chromatography (HPLC) diagram of the ginseng polysaccharide of example 1;
FIG. 2 is an elevated plus maze test; (a) number of times the rat entered the open arm in the test experiment; (B) sum of time to open arm;
FIG. 3 is an open field test; (A) The number of times a rat enters the open field central region in an open field test experiment; (B) a sum of times to enter the central zone;
FIG. 4 is a rat epileptiform behavioral test (limb/tail); (a) number of tics; (B) twitch duration;
fig. 5 is a rat epileptiform behavioral test (generalized tics); (a) systemic imbalance time; (B) number of systemic imbalances;
FIG. 6 is intestinal flora composition (portal level);
FIG. 7 is intestinal flora composition (genus level, top 10);
FIG. 8 is intestinal flora composition (seed level, top 10);
FIG. 9 is intestinal flora diversity; (a) alpha diversity; (B) beta diversity;
FIG. 10 is a differential analysis of intestinal flora; (A) linear discriminant analysis of LefSe; (B) Metastat non-parametric test analysis;
FIG. 11 is a graph of intestinal flora differentiation analysis, random forest and nonparametric test analysis.
Detailed Description
The invention provides a preparation method of ginseng polysaccharide, which comprises the following steps: pulverizing Ginseng radix, reflux-extracting with hot water, concentrating, centrifuging, precipitating supernatant with ethanol solution, dialyzing the precipitate with distilled water, and lyophilizing non-dialysate to obtain ginseng polysaccharide; the molecular weight cut-off of the dialysis was 3.5kDa.
In the invention, the ratio of the feed liquid extracted by hot water reflux is preferably 8-10 g/mL, more preferably 9g/mL; the temperature is preferably 100℃and the time is preferably 2 hours, the number of times is preferably 1 to 3, more preferably 2. The purpose of the hot water reflux extraction is to extract water-soluble components of ginseng.
In the present invention, the rotational speed of the centrifugation is preferably 2500 to 3500rpm, more preferably 3000rpm, and the time is preferably 20 to 40min, more preferably 30min.
In the invention, the volume percentage of the ethanol solution is preferably 75-85%, more preferably 80%; the volume ratio of the supernatant to ethanol solution extraction is preferably 1:4. The invention uses the method that the impurity is insoluble in ethanol and is settled at the bottom, and the purpose of removing the non-effective components is achieved by separating the supernatant.
The invention also provides ginseng polysaccharide obtained by the preparation method; the molecular weight of the ginseng polysaccharide is 627kDa.
The invention also provides application of the ginseng polysaccharide in preparing a product for regulating intestinal flora.
The invention also provides application of the ginseng polysaccharide in preparation of antidepressant drugs.
The invention also provides application of the ginseng polysaccharide in preparing medicines for treating epilepsy.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Preparation of ginseng polysaccharide
Pulverizing 300g dried Ginseng radix, reflux-extracting with 9 times of 100deg.C hot water for 2 hr, mixing the extracts, vacuum concentrating at 60deg.C to 40% of original volume, and centrifuging at 3000rpm for 30min to remove insoluble substances.
Precipitating the supernatant with 4 times of 80% ethanol solution, dissolving the precipitate in water, dialyzing with distilled water (molecular weight cut-off of 3.5 kDa), and lyophilizing the non-dialyzate to obtain ginseng polysaccharide.
Example 2
Preparation of ginseng polysaccharide
Pulverizing 300g dried Ginseng radix, reflux-extracting with 8 times volume of 100deg.C hot water for 2 hr, extracting for three times, mixing the extracts, concentrating under reduced pressure at 60deg.C to 40% of original volume, centrifuging at 3500rpm for 20min, and removing insoluble substances.
Precipitating the supernatant with 4 times of 75% ethanol solution, dissolving the precipitate in water, dialyzing with distilled water (molecular weight cut-off of 3.5 kDa), and lyophilizing the non-dialyzate to obtain ginseng polysaccharide.
Example 3
Preparation of ginseng polysaccharide
300g of dried ginseng root is crushed, 10 times volume of 100 ℃ hot water is used for reflux extraction for 2 hours, the extract is concentrated to 40% of the original volume under reduced pressure and vacuum at 60 ℃, and then centrifugation is carried out at 2500rpm for 40 minutes, so as to remove insoluble substances.
Precipitating the supernatant with 4 times of 85% ethanol solution, dissolving the precipitate in water, dialyzing with distilled water (molecular weight cut-off of 3.5 kDa), and lyophilizing the non-dialyzate to obtain ginseng polysaccharide.
Example 4
The polysaccharide obtained in example 1 was tested by High Performance Liquid Chromatography (HPLC)
Preparation of control solution
Weighing about 3mg of Pullulan 8 reference substances (5-800 kDa) powder, and adding 0.1 MNO respectively 3 1ml, heating to dissolve, centrifuging (13000 r.p.for 10 min), and collecting supernatant.
Preparation of test solution
About 5mg of polysaccharide sample powder of example 1 was weighed and 0.1M NaNO was added 3 1ml, dissolved by heating, centrifuged (13000 rpm, 10 min)Clock), and taking the supernatant.
HPLC-RID conditions
High Performance Liquid Chromatography (HPLC) profile Agilent 1200HPLC-RID, thermoAcclaim SEC(5 μm, 7.8X100 mm), mobile phase 0.1M NaNO 3 Isocratic elution, flow rate 0.8ml/min, column temperature: 30 ℃, sample injection amount: 10 μl, detector: RID (positive mode). A High Performance Liquid Chromatography (HPLC) diagram is shown in FIG. 1.
FIG. 1 shows a main peak having a molecular weight of 627kDa, and it can be seen that the ginseng polysaccharide prepared in example 1.
Example 5
Influence of panaxan on anxiety and depression behavior of mice
Wild type SD rats (360-420 g) were selected and divided into GP group (7) and Control group (6). The ginseng polysaccharide GP prepared in example 1 was dissolved in physiological saline (10 mg/mL) and stored in a refrigerator at 4℃for use. The experimental animals purchase Zhejiang Weitong Lihua and after 1-2 weeks of feeding room adaptation, the gastric lavage experiment is started. The body weight of the rats is weighed before the experiment, the rats of the experiment group are irrigated with (GP, 50 mg/kg), and the rats of the control group are irrigated with normal saline with the same amount of the corresponding body weight (the operation of the lavage is carried out at the time of 2-3 pm every day). The behavioural test was prepared after 6 days of continuous gavage, and the behavioural test was performed 2 hours after the end of the gavage test.
5.1 overhead laboratory test (Elevated plus maze, EPM)
5.1.1 test methods
The laboratory was dimly lit (based on the lowest brightness that allowed the rat to be distinguished from minute movements at a distance of 1.5 m) and kept constant bright at room temperature of 21 c and quiet. The test box was placed in one corner of the laboratory with a 2m high black monotonous background surrounding the test box. Only one person involved in daily handling carried the animals during the experiment.
Before experimental test, each rat is put into a 60cm X60cm X35cm plastic box, and after free exploration for 5min, the rat is quickly placed at the central platform of the four-wall maze to enable the head of the rat to face one of the open walls, and after release, the following indexes are recorded: (1) open wall entry (count): the number of times of entering any open arm is based on that four paws of a rat enter the arm, and the complete withdrawal of one paw from the arm in the middle is the completion of the entering activity; (2) open arm time sum (second): time to open arm, unit: second.
5.1.2 experimental results
The results of the behavioral tests (elevated plus maze) of the rats are shown in figure 2. Compared with Control group rats, the residence times and the residence time of the rats given with GP in the open arms are obviously increased, which indicates that the GP can prevent and relieve anxiety-depression-like behaviors of the rats.
5.2 Open Field Test (OFT)
5.2.1 test methods
After the experimental device is installed, the rats are taken out of the feeding cage, are placed back to the experimenter to the central area of the mine, and the experimenter quickly leaves, so that the rats can freely move in the experimental box. The recording software Supermaze was started quickly and the animal was recorded for 3-5min of activity in the mine. And after the experimental monitoring time is over, taking out the rat and putting the rat back into the rearing cage, and thoroughly cleaning the bottom, the placed matter and the side wall of the mine reaction box so as to prevent the information (such as the urine and the smell of the animal) remained in the animal from affecting the next test result. The whole open field box body is cleaned by 75% alcohol, and the experiment is continuously repeated after the rats are replaced after the rats are dried in the air.
5.2.2 experimental results
The results of the behavioral tests (open field experiments) of rats are shown in FIG. 3. The number of stay and time of rats given GP in the central region were significantly increased compared to Control group rats, indicating that GP can prevent and alleviate anxiety-depression-like behavior in rats.
Example 6
Influence of panaxan on mouse epileptic behaviour
6.1 test method
Wild type SD rats (360-420 g) were selected and divided into GP group (7) and Control group (6). The ginseng polysaccharide GP prepared in example 1 was dissolved in physiological saline (10 mg/mL) and stored in a refrigerator at 4℃for use. The experimental animals purchase Zhejiang Weitong Lihua and after 1-2 weeks of feeding room adaptation, the gastric lavage experiment is started.
The body weight of the rats is weighed before the experiment, the rats of the experiment group are irrigated with (ginseng polysaccharide GP,50 mg/kg), and the rats of the control group are irrigated with physiological saline with the same amount of the corresponding body weight (the operation of the lavage is carried out at the time of 2-3 pm every day). The behavioural test was prepared 7-8 days after the continuous gavage, and the behavioural test was performed 2 hours after the end of gavage.
Dorsal hippocampal CA3 position was determined by stereotactic, and fixed-point administration of alginic acid (kainic acid, KA,0.3mg/ml, equivalent to 0.375 nL/g) by microinjection induced epileptic development. Continuously observing and recording the seizure process of the rat, and counting the number and time of the twitches of the limbs/tails; systemic tic imbalance times and times.
6.2 test results
The comparison results of seizure process of rats are shown in fig. 4-5. Compared with Control group rats, the number and time of limb/tail twitches and the systemic twitch imbalance time and number of times of the rats given with GP are obviously reduced, which indicates that GP can prevent and relieve epileptiform behaviors of the rats.
Intestinal flora has been shown to be in significant association with depression, the development of epilepsy. The directions of application of intestinal flora in depression and epilepsy include: 1) The intestinal flora is regulated by fecal microorganisms or probiotics and the like, and can be used as an auxiliary means for treating or preventing depression and epilepsy. 2) The intestinal flora detection can be used as a biomarker for diagnosis or prognosis evaluation. 3) The intestinal flora analysis is used for guiding individual medication, the drug effect and the safety are improved, and the specific details are shown in example 7. Therefore, the ginseng polysaccharide prepared by the invention has the effect of treating depression and epilepsy.
Example 7
Intestinal flora detection and analysis method
7.1 test method
Faeces of mice of Control and GP groups after epileptic behavioural experiments in example 6 were subjected to intestinal microbiota detection: microorganism DNA in the fecal sample is extracted and amplified by using a kit matched with Shorelin Biome company, and a sequencing library is constructed after quality control. Sequencing was performed based on the PacBio sequence three-generation sequencing platform and the 16S-ITS-23S full-length amplicon method sequencing protocol. After resolution and quality control of the raw sequencing run data, highly accurate Amplified Sequence Variants (ASV) were deduced using DADA2, and then species annotation was performed at the phylum, class, order, family, genus, species taxonomic level. Based on QIIME2 platform and R analysis kit, further flora composition and structure analysis was performed to compare the relative abundance profile of intestinal flora before and after treatment. The flora composition and the structural difference among groups are evaluated through Beta diversity analysis (Beta diversity), the Bray-curtis distance among samples is calculated firstly, and then principal coordinate analysis (PCoA), principal Component Analysis (PCA) and non-metric multidimensional scale analysis (NMDS) are carried out; differential analysis was performed using Metastat, ANCOM, LEfSe and random forest algorithms to find potential biomarkers between different groupings; functional predictive analysis was performed using PICRUSt2 to investigate in depth colony function in different samples.
7.2 test results
The results of intestinal flora are shown in figures 6-10.
Comparing the top 6 species of relative abundance ranking at the gate, genus and species levels, the results show that GP group increased the average of the relative abundance of bacterioides and decreased the average of the relative abundance of Firmicutes compared to Control group (fig. 6). Meanwhile, at the genus level, the average of the relative abundance of bacterioides and turicibacillus was higher in GP group than in Control group (fig. 7). The composition relative abundance plot at the species classification level can be seen that the GP group, lactobacillus johnsonii, bacteroides co-protocola, was slightly elevated and the tunebacillus murinus, lactobacillus _ reuteri, akkermansia _muciniti was reduced compared to the Control group (fig. 8).
The NMDS results of β diversity showed stress <0.2, with some differences in flora structure between the two groups, indicating that GP has some intestinal flora modulating function (fig. 9). Two other methods of evaluating the two sets of samples, PCoA, PCA, also showed that the two sets of data were well-differentiated. Further studies of GP regulating intestinal flora process, differential analysis using Metastat showed that lactobacillus_ johnsonii, bacteroides _coporola and turicibacillus_sanguinis could be potential biomarkers (fig. 10). Significant features at seed level were calculated using random forests and nonparametric tests, and the results also validated the biomarker species previously shown (fig. 11).
Among them, lactobacillus johnsonii has been reported in the literature as an intestinal probiotic having anti-inflammatory, intestinal barrier repairing effects, and mental activity improving effects on mice dysmnesia [ Wang H, sunY, xin J, zhang T, sun N, ni X, zeng D, bai Y.Lactobacillus johnsonii BS15Prevents Psychological Stress-Induced Memory Dysfunction in Mice by Modulating the Gut-BrainAxis. Front Microbiol 13 (11) (2020) 1941 ]; bifidobacteria and lactobacilli are lower than 4 times per year in patients with 4 times per year of episodes or more [ Peng, a., qiu, x., lai, w., li, w., zhang, l., zhu, x., he, s, dutan, j., and Chen, l., altered composition of the gut microbiome in patients with drug-resistance epiepsy.epiepsy Res 147 (2018) 102-107 ]. Bactoides_copoloa reported that the encoded glycosidase gene has a correlation with T2D [ ChenY, li Z, hu S, zhang J, wu J, shao N, bo X, ni M, ying X.Gut metagenomes of type 2diabetic patients have characteristic single-nucleotide polymorphism distribution in Bacteroides copoloa. Microome 5 (1) (2017) 15 ]; turicibacter_sanguinis has been reported to improve depression by increasing serotonin (5-hydroxytryptamine) in the gut [ Fung TC, vuong HE, luna CDG, pronovest GN, aleks androva AA, riley NG, vavilinaA, mcGinn J, rendon T, forrest LR, hsiao EY. Interal serotonin and fluoxetine exposure modulate bacterial colonization in the gun. Nat Microbiol4 (12) (2019) 2064-2073.1 ].
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A method for preparing ginseng polysaccharide, which is characterized by comprising the following steps: pulverizing Ginseng radix, reflux-extracting with hot water, concentrating, centrifuging, precipitating supernatant with ethanol solution, dialyzing the precipitate with distilled water, and lyophilizing non-dialysate to obtain ginseng polysaccharide; the molecular weight cut-off of the dialysis was 3.5kDa.
2. The preparation method according to claim 1, wherein the ratio of the feed liquid extracted by hot water reflux is 8-10 g/mL, the temperature is 100 ℃, the time is 2h, and the times are 1-3 times.
3. The method according to claim 1, wherein the rotational speed of the centrifugation is 2500-3500 rpm for 20-40 min.
4. The method according to claim 1, wherein the ethanol solution is 75-85% by volume.
5. The method according to claim 1, wherein the volume ratio of the supernatant to the ethanol solution extraction is 1:4.
6. A ginseng polysaccharide obtained by the process according to any one of claims 1 to 5.
7. The ginseng polysaccharide according to claim 6, wherein the molecular weight of the ginseng polysaccharide is 627kDa.
8. Use of ginseng polysaccharide according to claim 6 or 7 for the preparation of products for regulating intestinal flora.
9. Use of the ginseng polysaccharide according to claim 6 or 7 for the preparation of antidepressants.
10. Use of ginseng polysaccharide according to claim 6 or 7 for the preparation of a medicament for the treatment of epilepsy.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698684A (en) * | 2009-10-23 | 2010-04-28 | 武汉大学 | Method for extracting polysaccharose from ginseng and application |
| CN116589604A (en) * | 2023-06-05 | 2023-08-15 | 广东省科学院江门产业技术研究院有限公司 | Preparation method of high-purity ginseng polysaccharide |
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698684A (en) * | 2009-10-23 | 2010-04-28 | 武汉大学 | Method for extracting polysaccharose from ginseng and application |
| CN116589604A (en) * | 2023-06-05 | 2023-08-15 | 广东省科学院江门产业技术研究院有限公司 | Preparation method of high-purity ginseng polysaccharide |
Non-Patent Citations (4)
| Title |
|---|
| WANG J等: "Antidepressant-like effects of the active acidic polysaccharide in mice", JOURNAL OF ETHNOPHARMACOLOGY, vol. 132, no. 01, 29 July 2010 (2010-07-29), pages 65 - 69 * |
| 刘鹏飞;: "人参多糖穴位注射辅助治疗抑郁障碍的临床对照研究", 内蒙古中医药, no. 10, 30 May 2017 (2017-05-30), pages 103 * |
| 祁玉丽: "人参多糖对肠道微生态及肠黏膜免疫作用的研究", 中国优秀硕士学位论文全文数据库医药卫生科技辑, no. 03, 15 March 2020 (2020-03-15), pages 057 - 255 * |
| 郭力 等: "中药化学", vol. 02, 31 August 2018, 中国医药科技出版社, pages: 17 * |
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