A kind of coralline hericium erinaceus polysaccharide and its preparation method and application

Abstract

This invention belongs to the field of pharmaceutical technology, specifically relating to a coral-shaped Hericium erinaceus polysaccharide, its preparation method, and its application. The dosage of the coral-shaped Hericium erinaceus polysaccharide is 50 mg / kg, and its molecular weight is 16.06 kDa. The coral-shaped Hericium erinaceus polysaccharide is mainly composed of three monosaccharides: galactose, fucose, and mannose, with a molar ratio of 83.931:14.516:1.102. It has neuroprotective effects, can alleviate Alzheimer's disease symptoms, and improve related pathological indicators in the brain. It can be used to prepare neuroprotective and Alzheimer's disease prevention and treatment drugs and health products.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a coral-like Hericium erinaceus polysaccharide, its preparation method, and its application. Background Technology

[0002] Alzheimer's disease (AD) causes a decline in memory and other cognitive abilities, and is one of the most serious diseases in the field of geriatrics. According to research reports, as of 2021, more than 50 million people worldwide suffer from AD, resulting in economic losses of up to $100 billion. Since its discovery more than 100 years ago, researchers have proposed several hypotheses regarding the pathogenesis of AD, including the β-amyloid cascade hypothesis, the abnormal phosphorylation hypothesis of Tau protein, the neuronal apoptosis hypothesis, the cholinergic damage hypothesis, the metal ion hypothesis, and the immune-inflammatory theory. However, developing drugs targeting its underlying mechanisms has proven extremely difficult.

[0003] Currently, clinical diagnosis of Alzheimer's disease (AD) primarily relies on behavioral symptoms. By this time, most cases are diagnosed in the middle to late stages, with impaired cognitive function and progressive cognitive impairment or behavioral changes. AD is progressive and irreversible, and there is currently no specific cure. Most medications available for mild to moderate AD are suitable for patients with the disease. Common medications include galantamine hydrobromide, donepezil hydrochloride, rivastigmine, and memantine hydrochloride. However, these medications are relatively expensive, have significant side effects, and can only slow disease progression, not reverse it.

[0004] Hericium coralloides, also known as the coral-shaped monkey head mushroom, is a famous and rare edible fungus in my country, mainly distributed in the three northeastern provinces of my country, as well as Sichuan, Yunnan, Xinjiang, and Tibet. It is rich in protein, polysaccharides, trace elements, and amino acids. Studies have shown that Hericium coralloides polysaccharides have strong antioxidant activity and exhibit a certain ability to lower cholesterol. Ethyl acetate extract of Hericium coralloides increased the density of cortical neurons in D-galactose mice, improving aging conditions. However, there is currently no research on the use of Hericium coralloides polysaccharides in preventing Alzheimer's disease. Summary of the Invention

[0005] The purpose of this invention is to provide a coral-like Hericium erinaceus polysaccharide, its preparation method, and its application. Using Hericium erinaceus fruiting bodies as raw materials, a coral-like Hericium erinaceus polysaccharide is obtained through extraction, separation, and purification. Its structural characterization, neuroprotective activity, and activity in preventing and treating Alzheimer's disease are then investigated.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] This invention provides a coral-like Hericium erinaceus polysaccharide with a molecular weight of 16.06 kDa. The coral-like Hericium erinaceus polysaccharide is mainly composed of three monosaccharides: galactose, fucose, and mannose, with a molar ratio of 83.931:14.516:1.102.

[0008] Furthermore, the specific structure of the coral-like Hericium erinaceus polysaccharide is as follows:

[0009] →6)-α-D-Galp-(1→ serves as the main chain and undergoes partial methylation at its O-3 position, while at the O-2 position of →6)-α-D-Galp-(1→, it is partially replaced by α-L-Fucp-(1→).

[0010] The coral-like Hericium erinaceus polysaccharide is composed of 5 glycoside fragments, wherein the molar proportions of the glycoside fragments T-Fucp, 6-Galp and 2,6-Galp are 17.178%, 52.257% and 25.277%, respectively.

[0011] The present invention also provides a method for preparing the aforementioned coral-like Hericium erinaceus polysaccharide, comprising the following steps:

[0012] (1) The coral-shaped Hericium erinaceus fruiting bodies were ultra-finely pulverized into powder and passed through an 80-mesh sieve to obtain fungal powder;

[0013] (2) Add deionized water to the bacterial powder, extract, and then combine the solutions to obtain the extracted solution;

[0014] (3) Centrifuge the solution extracted in step (2) to collect the supernatant, then concentrate it by rotary evaporation, perform alcohol precipitation treatment, incubate at 4°C overnight, and then centrifuge to collect the precipitate.

[0015] (4) Dissolve the precipitate obtained in step (3) again with deionized water, remove the protein by Sevag method, remove the residual organic solvent by dialysis, evaporate to a suitable concentration and freeze dry to obtain coral-like Hericium erinaceus crude polysaccharide.

[0016] (5) Dissolve the crude polysaccharide of Hericium erinaceus in deionized water, load it onto a DEAE-52 anion exchange column and elute with deionized water, collect it in a test tube, concentrate the collected Hericium erinaceus polysaccharide to a suitable concentration by rotary evaporation, and then lyophilize it by dialyzing to obtain Hericium erinaceus polysaccharide.

[0017] (6) The coral-like Hericium erinaceus polysaccharide lyophilized powder obtained by passing through a DEAE-52 ion exchange column was dissolved in deionized water and loaded onto Sephacryl. TM S400 column, after dialysis, rotary evaporation, and freeze-drying to obtain coral-like Hericium erinaceus S400;

[0018] (7) The coral-shaped Hericium erinaceus S400 was dissolved in deionized water, loaded onto a Chromdex 200pg column, dialyzed, and then evaporated by rotary evaporation and freeze-dried to obtain Hericium erinaceus polysaccharide (HCP).

[0019] Furthermore, in step (2), the amount of deionized water added is 30 times the mass of the bacterial powder; the extraction is carried out in a constant temperature water bath at 80°C for 2 hours, and the extraction is repeated twice.

[0020] Furthermore, in step (3), the centrifugation is performed at 8000 rpm for 10 minutes; the rotary evaporation is concentrated to 1 / 10 of the volume; and the alcohol precipitation is performed by adjusting the final ethanol concentration to 80% (v / v).

[0021] Furthermore, in step (6), the sample is loaded onto Sephacryl... TM The eluent after the S400 column was deionized water, and the flow rate was 0.8 mL / min; the dialysis time was 72 h.

[0022] Furthermore, in step (7), the eluent after loading the sample onto the Chromdex 200pg column is deionized water, and the flow rate is 0.8mL / min; the dialysis time is 72h.

[0023] The present invention also provides the application of the aforementioned coral-like Hericium erinaceus polysaccharide in the preparation of drugs for the prevention and treatment of Alzheimer's disease.

[0024] The present invention also provides the use of the aforementioned Hericium erinaceus polysaccharide in the preparation of neuroprotective drugs.

[0025] The present invention also provides a pharmaceutical formulation, wherein the aforementioned Hericium erinaceus polysaccharide is the active ingredient, and the formulation contains one or more pharmaceutically acceptable carrier substances and / or excipients.

[0026] The beneficial effects of this invention are as follows:

[0027] A polysaccharide of Hericium erinaceus with high neuroprotective activity is provided. It has neuroprotective effects, can alleviate Alzheimer's disease symptoms and improve related biochemical indicators in the brain, and can be used to prepare drugs and health products for neuroprotection and prevention of Alzheimer's disease. Attached Figure Description

[0028] Figure 1 Elution curve of coral-like Hericium erinaceus polysaccharide DEAE-52;

[0029] Figure 2 Sephacryl (a polysaccharide from Hericium erinaceus) TM S400 elution curve;

[0030] Figure 3 Elution curve of Chromdex polysaccharide from Hericium erinaceus (coral-shaped) at 200 pg;

[0031] Figure 4 Infrared spectrum of Hericium erinaceus polysaccharide;

[0032] Figure 5 : Monosaccharide composition analysis of Hericium erinaceus polysaccharide;

[0033] Figure 6 NMR of coral-like Hericium erinaceus polysaccharide 1 H spectrum;

[0034] Figure 7 NMR of coral-like Hericium erinaceus polysaccharide 13 C spectrum;

[0035] Figure 8 : Results of mouse jumping on the platform;

[0036] Figure 9 Results of the mouse water maze test;

[0037] Figure 10 Aβ in the mouse hippocampus 42 Immunohistochemistry results image;

[0038] Figure 11 Immunohistochemical results of Tau (phospho S396) in the hippocampus of mice. Detailed Implementation

[0039] The present invention will be further described below with reference to the embodiments. These embodiments are merely exemplary and do not constitute any limitation on the scope of the present invention. Those skilled in the art should understand that modifications or substitutions can be made to the details and form of the technical solutions of the present invention without departing from the spirit and scope of the present invention, but such modifications and substitutions all fall within the protection scope of the present invention. Unless otherwise specified, the chemical reagents, chromatography columns, etc. used in the specification and embodiments shall be operated under conventional experimental conditions or according to the instructions provided by the supplier.

[0040] This invention yields a novel coral-like Hericium erinaceus polysaccharide. Using Hericium erinaceus fruiting bodies as raw material, crude polysaccharide was obtained through hot water extraction, alcohol precipitation, concentration, and freeze-drying. The crude polysaccharide was then subjected to DEAE-52 and Sephacryl... TM Coral-shaped Hericium erinaceus polysaccharide was obtained by lyophilization with S400 and Chromdex 200 pg. The structure of the Hericium erinaceus polysaccharide was characterized by infrared spectroscopy, gel permeation chromatography, GC-MS, and nuclear magnetic resonance. Its neuroprotective effects and prevention of Alzheimer's disease were investigated through animal experiments.

[0041] The coral-shaped Hericium erinaceus fruiting bodies used in this invention were obtained through cultivation, and the strain number is 2020071401.

[0042] Example 1:

[0043] The preparation method of coral-like Hericium erinaceus polysaccharide is as follows:

[0044] 1) The coral-shaped Hericium erinaceus fruiting bodies are ultra-finely pulverized into powder and passed through an 80-mesh sieve;

[0045] 2) Add 30 times the weight of the bacterial powder to deionized water, extract in a constant temperature water bath at 80℃ for 2 hours, extract twice, and combine the solutions;

[0046] 3) After cooling, centrifuge at 8000 rpm for 10 minutes, take the supernatant, concentrate it to 1 / 10 volume by rotary evaporation, adjust the final ethanol concentration to 80% (v / v) for alcohol precipitation, incubate at 4℃ overnight, and collect the precipitate by centrifugation.

[0047] 4) Dissolve the precipitate again with deionized water, remove the protein using the Sevag method, remove the residual organic solvent by dialysis, evaporate to the appropriate concentration and freeze dry to obtain coral-like Hericium erinaceus crude polysaccharide (HCCP).

[0048] 5) Dissolve HCCP in deionized water, load the sample onto a DEAE-52 anion exchange column for elution, and collect the 8 mL sample in a clean, dry test tube. Detect the collected sugar using the phenol-sulfuric acid method according to different eluent concentrations. Plot the elution curves and name them HCCP-N (0 M NaCl), HCCP-1 (0.1 M NaCl), HCCP-2 (0.3 M NaCl), and HCCP-3 (0.5 M NaCl), respectively. Concentrate the HCCP to a suitable concentration by rotary evaporation, dialyze for 72 h, and freeze-dry to obtain coral-like Hericium erinaceus polysaccharide (HCCP-N).

[0049] 6) Dissolve HCCP-N obtained by passing through a DEAE-52 ion exchange column in deionized water and load the solution onto a Sephacrylene plate. TM An S400 column (2.6 cm × 60 cm) was used with deionized water as the eluent at a flow rate of 0.8 mL / min. The automatic collector was set to a volume of 5 mL / tube. The polysaccharide content was determined using the phenol-sulfuric acid method. The main peak was collected, dialyzed for 72 h, rotary evaporated to 1 / 5 of the original volume, and then lyophilized to obtain coral-like Hericium erinaceus S400 (HCCP-S400).

[0050] 7) Will be via Sephacryl TMHCCP-S400 on an S400 column was dissolved in deionized water and loaded onto a Chromdex 200 pg column (2.6 cm × 60 cm). Deionized water was used as the eluent, and the flow rate was 0.8 mL / min. The automatic collector was set to a volume of 2 mL / tube. The polysaccharide content was determined by the phenol-sulfuric acid method. The main peak was collected, dialyzed for 72 h, rotary evaporated to 1 / 5 of the original volume, and lyophilized to obtain coral-like Hericium erinaceus polysaccharide (HCP).

[0051] Test Example 1: HCP Structural Feature Analysis

[0052] (1) Test materials

[0053] Example 1: HCP was obtained.

[0054] (2) Experimental methods and results

[0055] Infrared spectroscopy analysis: The HCP obtained in Example 1 was mixed with potassium bromide (mass ratio 1:200) dried under microwave conditions at 100°C and compressed into a tablet, then subjected to infrared spectroscopy scanning. The scanning results are as follows: Figure 4 As shown, in the range of 500–4000 cm -1 Within the scanning wavelength range, HCP is at 3430 cm⁻¹. -1 The peak at 2919 cm⁻¹ is the absorption peak of the OH stretching vibration, a characteristic peak of carbohydrates. -1 There is an absorption peak at 1641 cm⁻¹, which is attributed to the CH stretching vibration. -1 The absorption peak at 1078 cm⁻¹ is caused by the stretching vibration of C=O. -1 The absorption peak at 1000-1200 cm⁻¹ indicates the presence of the pyranose ring; in addition, the absorption peak at 1000-1200 cm⁻¹ indicates the presence of the pyranose ring. -1 This is attributed to the tensile vibrations of COC and COH;

[0056] Monosaccharide component analysis: Accurately weigh 20.0 mg of HCP, dissolve it in 5 mol / L sulfuric acid, and hydrolyze it under sealed conditions at 105℃ for 6 h. After cooling, adjust the pH to 7.0 with barium carbonate. Centrifuge and filter the supernatant through a 0.22 μm filter membrane. Analyze by high-performance liquid chromatography (HPLC) / evaporative light scattering detector. An LC-10ATVP HPLC system with an Alltech 2000ES evaporative light scattering detector was used. The chromatographic column was a Prevail™ ES sugar column (250 × 4.6 mm), the mobile phase was 80% acetonitrile, the flow rate was 1.0 mL / min, the column temperature was 35℃, the drift tube temperature was 82℃, the carrier gas flow rate was 2.1 L / min, and the injection volume was 20 μL. The results are as follows: Figure 5 As shown, HCP is mainly composed of three monosaccharides, and their types and molar ratios are: D-galactose (Gal): L-fucose (Fuc): D-mannose (Man) = 83.931: 14.516: 1.102.

[0057] Methylation and acetylation analysis: To determine the bonding structure of the polysaccharide, methylation and acetylation reactions were performed. The prepared samples were analyzed by GC-MS. The derivative type was determined by its typical retention time and electron collision curve. The steps are as follows: Methylation reaction: The entire reaction was carried out at 20℃ under N2 protection. 10 mg of dried HCP and its sugar acid reduction product were added to a round-bottom flask, 2 mL of DMSO was added and sonicated to dissolve, and then 30 mg of dried NaOH powder was added and reacted for 3 h. 1 mL of CH3I was slowly added dropwise along the wall, and the reaction was carried out by sonication in the dark for 2 h. The experiment was carried out in an ice-water bath. At the end of the reaction, 3 mL of water was added to terminate the reaction. CHCI3 was added in an equal proportion for extraction, and the organic phase was collected. This was repeated 3 times. After drying, methylation was repeated until no hydroxyl peaks were found in the IR scan spectrum. Subsequent reactions: 4 mL of 2 mol / L trifluoroacetic acid (TFA) was added to the methylated sample, and hydrolysis was carried out at 110 °C for 5 h. After hydrolysis, 4 mL of methanol was added and evaporated to remove excess TFA. This process was repeated 3 times. Then, 0.5 mL of distilled water was added to dissolve the TFA. 0.2 mL of the hydrolysate was taken and 30 mg of NaBH4 was added. The reaction was carried out at room temperature for 3 h with intermittent shaking during the reaction. After the reaction was completed, 25% glacial acetic acid was added dropwise until no bubbles were generated. 2 mL of methanol was added and evaporated to remove excess TFA. This process was repeated 3 times. 1.5 mL of acetic acid was added to the reduction product, and the reaction was carried out at 100 °C for 1 h. 2 mL of toluene was added and evaporated to remove excess acetic anhydride. 2 mL of chloroform and water were added to each of the acetylated products. The mixture was shaken, allowed to stand, and the lower organic phase was collected. The mixture was washed 3 times with 1 mL of water and then a small amount of anhydrous sodium sulfate was added to remove water. The mixture was then analyzed by GC-MS.

[0058] The chromatographic system used was an Agilent gas chromatograph (Agilent 7890A). The mass spectrometry system used was an Agilent quadrupole mass spectrometer (Agilent 5977B) with electron collision ionization (EI) and a MassHunter workstation. The chromatographic column was an HP-5MS capillary column (30m × 250pm × 0.25umD); the carrier gas was He; the heater temperature was 250℃; the programmed temperature rise was: initial temperature 140℃ / min to 200℃, hold for 5 min, then rise at 8℃ / min to 240℃; the split ratio was 50:1; the injection volume was 5 μL. The analyte was detected using EI in full scan mode, with a mass scan range (m / z) of 30-600. The binding method was confirmed by comparing characteristic fragments of methylated polysaccharides with existing databases.

[0059] Based on the retention time and standard data of partially methylated sugar alcohol acetates from the Spectral Database of the Center for Complex Carbohydrates, the Hericium erinaceus polysaccharide is mainly composed of five glycoside fragments (Table 1). The molar ratio of non-reducing ends of T-Fucp reaches 17.178%, indicating the presence of branched residues.

[0060] Table 1. Methylation analysis results of HCP obtained in Example 1

[0061]

[0062] Nuclear magnetic resonance (NMR) analysis: Take an appropriate amount of HCP obtained in Example 1, dissolve it in 0.5 ml of D2O, and perform NMR analysis using a Bruker 500 MHz NMR spectrometer. Figure 6 and 7 The nuclear magnetic resonance imaging of coral-like Hericium erinaceus polysaccharide is as follows: 1 H spectrum and 13 C-spectrum. In HCP 1 In H NMR, four anomeric hydrogen signals were observed in the δ range of 4.9–5.0 ppm, all >4.9 ppm, suggesting that all four sugar residues in HCP are α-phenotypes (Table 2). HCP's 1 H and 13 In C NMR, at δ H Four anodic hydrogen signals were detected at values ​​of 4.91, 5.0, 4.98, and 5.0, δ C Four anomeric carbon signals were found at values ​​of 96.81, 100.19, 96.76, and 100.40, corresponding to H1 / C1 of the sugar residue AD, respectively. 1 H- 1 H COSY spectra show that δ H / H There are six cross peaks at 4.91 / 3.78, 3.78 / 3.81, 3.81 / 3.94, 3.94 / 4.14, 4.14 / 3.6, and 4.14 / 3.85, which correspond to sugar residue A, δ H The H2-H6a / b signals at 3.78, 3.81, 3.94, 4.14, and 3.60 / 3.85 are consistent. Based on the above method, the BD sugar residues were analyzed, and the O-CH3 signal of sugar residue B was found at 3.38 / 55.00. The connection mode of HCP was inferred by combining heteronuclear single quantum relation (HSQC), multi-bonded carbon-hydrogen relation (HMBC), and hydrogen-hydrogen relation (COSY). The results are shown in Table 2. The structure can be represented as →6)-α-D-Galp-(1→ as the main chain and partially methylated at its O-3 position, with α-L-Fucp-(1→ partially substituted at the O-2 position of →6)-α-D-Galp-(1→).

[0063] Table 2 shows the HCP obtained in Example 1. 1 H and 13 C NMR chemical shift information

[0064]

[0065] Test Example 2: Detection of the neuroprotective effect of HCP

[0066] Test materials

[0067] The test drugs were HCP obtained in Example 1 and sterile water.

[0068] Animals: The animal experiments conducted in accordance with institutional guidelines were approved by the Animal Ethics Committee of Jilin Agricultural University (20211107001). Twenty-four B6C3-Tg(APPswePSEN1dE9) / Nju double transgenic male mice (APP / PS1) (genotype: (Appswe)T, (Psen1)T) and 12 age-matched wild-type (WT) male mice (8 weeks old, SPF grade) were purchased from Changzhou Cavens Laboratory Animal Co., Ltd. All mice were housed in an environment with a temperature of 23±1℃ and humidity of 50% to 60%. A 12-hour illumination period with uniform natural light was maintained, and the mice had free access to food and water.

[0069] Mouse grouping and administration: 8-month-old APP / PS1 mice were randomly divided into two groups of 12 mice each. After one week of acclimatization feeding, the APP / PS1 group was given sterile water by gavage. The administration group was given HCP 50 mg / kg obtained in Example 1. The blank group consisted of WT mice, which were given sterile water by gavage for 8 weeks.

[0070] Animal behavioral tests: Animal behavioral tests include the jumping platform test and the Morris water maze to examine the cognitive and memory abilities of HCP on APP / PS1 mice.

[0071] (1) The effect of HCP on short-term memory in APP / PS1 mice was investigated by jumping platform test.

[0072] Training began on day 45 of drug administration in WT and APP / PS1 mice. The animals were first placed in a jumping platform apparatus for 120 seconds to acclimatize, during which time no electricity was applied. After the acclimatization period, the power was turned on, supplying a 36V current to the entire platform. The mice received an electrical stimulus when they left the platform and touched the copper mesh; each electrical response was recorded as an incorrect response. This training was repeated for 300 seconds. Memory testing was then conducted 24 hours later, recording the time the mice spent on the platform (any time exceeding 300 seconds was recorded as 300 seconds). The results are as follows: Figure 8 As shown.

[0073] (2) The effects of HCP on learning and spatial memory in APP / PS1 mice were investigated using the Morris water maze test.

[0074] WT and APP / PS1 mice underwent water maze training starting on day 50 of drug administration, with a training period of 4 days. First, the water temperature was adjusted to 22±1℃, and titanium dioxide was used to whiten the water throughout the maze, with the water level approximately 1 cm above the platforms. The mice were then placed in the maze, and a timer was started. If a mouse failed to find a platform within one minute, it was guided to the platform using a tool and allowed to stay on it for 10 seconds. This was repeated 4 times daily (at least once from the other side of the platform; the other three quadrants were randomly placed). This process was repeated for 4 days to allow the mice to form a memory. On day 5, the time it took for the mice to find the platform was recorded using a water maze monitoring system. On day 6, the platforms were removed, and the number of times the mice traversed the platforms within 60 seconds was recorded using a water maze test. The results are as follows: Figure 9 As shown.

[0075] After the final behavioral test, the mice were fasted for 12 hours. Blood was collected and stored at -80°C. The mice were then euthanized by intraperitoneal injection of 100 mg / kg sodium pentobarbital. Brain tissue was rapidly collected.

[0076] Aβ in the brain 42 Tau (phospho S396) immunohistochemical staining: Brain tissue was fixed in 4% paraformaldehyde solution. The brain was sectioned into 20 μm slices at low temperature and washed twice with PBS for 5 min each time. The slices were then incubated in PBS solution with 3% hydrogen peroxide for 20 min to terminate endogenous peroxidase activity, followed by two more washes with PBS for 5 min each time. The brain slices were blocked in 5% BSA solution for 1 hour and incubated overnight at 4°C with the primary antibody. The antibody used and its dilution ratio were Aβ. 42 (Dilution ratio 1:300), Tau (phospho S396, dilution ratio 1:500). After incubation with the primary antibody, the brain sections were washed three times with PBS for 10 min each time. After washing, the brain sections were incubated with IgG secondary antibody (dilution ratio 1:500) at room temperature for 90 min. After staining with DAB staining solution for 10 min, the brain sections were dehydrated stepwise in the same grade of ethanol, cleared with xylene, mounted with neutral resin mounting medium, and examined under a light microscope.

[0077] (3) Test Results

[0078] Results of animal behavioral experiments:

[0079] Depend on Figure 8 It can be seen that in the jumping test, compared with the blank group, the APP / PS1 mice had a shorter latency period. After administration, the latency period increased, which proves that administration improved the memory of APP / PS1 mice.

[0080] Depend on Figure 9It can be seen that in the Morris water maze test, compared with the control group, the APP / PS1 mice had a significantly increased platform search time, proving their memory decline. HCP treatment significantly shortened the latency of APP / PS1 mice and increased the number of crossings, indicating that the long-term memory of AD mice was improved.

[0081] hippocampal Aβ 42 Tau (phospho S396) immunohistochemical staining results:

[0082] Excessive accumulation of Aβ and increased expression of p-Tau are two serious inducing factors in the pathological process of Alzheimer's disease (AD), which can significantly aggravate the progression of AD. Immunohistochemical analysis of the expression of related proteins in the mouse brain revealed that Aβ in the hippocampus... 42 The immunohistochemical results of Tau (phospho S396) are as follows: Figure 10 and Figure 11 Compared with APP / PS1 mice, HCP administration reduced Aβ. 42 Patch deposition ( Figure 10 It inhibited the amount of phosphorylated Tau protein in the mouse hippocampus. Figure 11 ).

[0083] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A coral-like Hericium erinaceus polysaccharide, characterized in that, The molecular weight of the coral-like Hericium erinaceus polysaccharide is 16.06 kDa. The coral-like Hericium erinaceus polysaccharide is mainly composed of three monosaccharides: galactose, fucose, and mannose, with a molar ratio of 83.931:14.516:1.

102. The specific structure of the coral-like Hericium erinaceus polysaccharide is as follows: →6)-α-D-Gal p -(1→ as the main chain and partially methylated in its O-3 phase, at →6)-α-D-Gal p -(1→ at O-2 part is α-L-Fuc p -(1→replace; The coral-like Hericium erinaceus polysaccharide consists of five glycoside fragments, with the major glycoside fragment being T-Fuc. p 6-Gal p and 2,6-Gal p The molar proportions were 17.178%, 52.257%, and 25.277%, respectively.

2. The method for preparing coral-like Hericium erinaceus polysaccharide as described in claim 1, characterized in that, Includes the following steps: (1) The coral-shaped Hericium erinaceus fruiting bodies are crushed into powder and passed through an 80-mesh sieve to obtain mycelium powder; (2) Add deionized water to the bacterial powder, extract, and then combine the solutions to obtain the extracted solution; (3) Centrifuge the solution extracted in step (2) to collect the supernatant, concentrate it by rotary evaporation, perform alcohol precipitation treatment, incubate at 4°C overnight, and collect the precipitate by centrifugation; (4) Dissolve the precipitate obtained in step (3) again with deionized water, remove the protein by Sevag method, remove the residual organic solvent by dialysis, evaporate to a suitable concentration and freeze dry to obtain coral-like Hericium erinaceus crude polysaccharide. (5) Dissolve the crude polysaccharide of Hericium erinaceus in deionized water, load it onto a DEAE-52 anion exchange column and elute with deionized water, collect it in a test tube, concentrate the collected Hericium erinaceus polysaccharide to a suitable concentration by rotary evaporation, and obtain neutral polysaccharide of Hericium erinaceus by dialysis and freeze drying. (6) The neutral polysaccharide powder of the coral-like Hericium erinaceus obtained by the DEAE-52 ion exchange column was dissolved in deionized water, and was loaded onto a Sephacryl TM S400 column. After dialysis, rotary evaporation, and lyophilization, the coral-like Hericium erinaceus S400 was obtained. (7) Dissolve Hericium erinaceus S400 in deionized water, load the sample onto a Chromdex 200 pg column, dialyze, and then evaporate by rotary evaporation and freeze-dry to obtain Hericium erinaceus polysaccharide.

3. The method for preparing coral-like Hericium erinaceus polysaccharide according to claim 2, characterized in that, In step (2), the amount of deionized water added is 30 times the mass of the bacterial powder; the extraction is carried out in a constant temperature water bath at 80℃ for 2 hours, and the extraction is repeated twice.

4. The method for preparing coral-like Hericium erinaceus polysaccharide according to claim 2, characterized in that, In step (3), the centrifugation is performed at 8000 rpm for 10 minutes; the rotary evaporation is concentrated to 1 / 10 of the volume; and the alcohol precipitation is performed by adjusting the final ethanol concentration to 80% by volume.

5. The method for preparing coral-like Hericium erinaceus polysaccharide according to claim 2, characterized in that, In step (6), the sample was loaded on Sephacryl TM The eluent after S400 column was deionized water at a flow rate of 0.8 mL / min; the dialysis time was 72 h.

6. The method for preparing coral-like Hericium erinaceus polysaccharide according to claim 2, characterized in that, In step (7), the eluent after loading the sample onto the Chromdex 200 pg column is deionized water, and the flow rate is 0.8 mL / min; the dialysis time is 72 h.

7. The application of the coral-like Hericium erinaceus polysaccharide according to claim 1 in the preparation of drugs for the prevention and treatment of Alzheimer's disease.

8. The use of the Hericium erinaceus polysaccharide according to claim 1 in the preparation of neuroprotective drugs.

9. A pharmaceutical preparation, characterized in that, The pharmaceutical preparation uses the coral-like Hericium erinaceus polysaccharide of claim 1 as the active ingredient, and also contains one or more pharmaceutically acceptable carrier substances and / or excipients.

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