Bulbus fritillariae ussuriensis polysaccharide extract for preventing lung cancer as well as preparation method and application thereof

Abstract

The invention discloses bulbus fritillariae ussuriensis polysaccharide as well as a preparation method and application thereof in lung cancer resistance, and belongs to the technical field of extraction of effective components of traditional Chinese medicines. According to the method, polysaccharide components in bulbus fritillariae ussuriensis bulbs are extracted, the extract is subjected to impurity removal through membrane separation, deproteinization is conducted through a macroporous resin method, the sugar content of the polysaccharide is measured, monosaccharide composition analysis is conducted through an HPLC method, and structural characterization is conducted on the obtained bulbus fritillariae ussuriensis polysaccharide through UV, IR and 1H-NMR. Finally, in-vivo and in-vitro researches find that the fritillary bulb polysaccharide has the effect of treating and preventing lung cancer, so that the invention provides a theoretical basis for further in-depth researches of fritillary bulb polysaccharide components and medicinal activity, and meanwhile, the invention combines chemical components, drug effects and action mechanisms, and expands natural plant resources of anti-cancer drugs.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of effective component extract of traditional Chinese medicine, and particularly relates to a Fritillariae Pallidicaulis Bunge polysaccharide extract for preventing lung cancer and a preparation method and application thereof. BACKGROUND

[0002] Pulmonary infectious diseases, including pneumonia, tuberculosis, bronchiectasis, etc. are direct factors causing lung cancer. The morbidity and mortality of lung cancer are at the top of global cancers, and are increasing year by year. In China, lung cancer has ranked the first in the list of death causes of urban malignant tumors.

[0003] Fritillariae Pallidicaulis Bunge is the dried bulb of Fritillariae Pallidicaulis Bunge of Liliaceae. Fritillaria ussuriensis It is bitter, sweet and cold in nature, and belongs to the lung and heart meridians. It has the functions of clearing heat and moistening the lungs, reducing phlegm and relieving cough. It is mainly used for lung heat and dry cough, dry cough with little phlegm, yin deficiency and cough with blood-stained sputum. The chemical components of Fritillariae Pallidicaulis Bunge mainly include alkaloids, polysaccharides, etc. Existing studies have shown that Fritillariae Pallidicaulis Bunge polysaccharide may be the main component of Fritillariae Pallidicaulis Bunge. In addition, Fritillariae Pallidicaulis Bunge has the effects of treating lung fire and lung heat and dry cough.

[0004] Polysaccharides are mainly widely used in medicine. However, it is difficult to extract and separate due to its complex structure. At present, the research on Fritillariae Pallidicaulis Bunge polysaccharide mainly focuses on physiological activity and extraction process, and there are few reports on the separation and purification process and structural characteristics of Fritillariae Pallidicaulis Bunge polysaccharide. The chemical structure and efficacy are not combined, so it cannot play the role of antioxidant and prevention of lung cancer, which seriously limits the development and application of Fritillariae Pallidicaulis Bunge. SUMMARY

[0005] The present application provides a preparation method and application of Fritillariae Pallidicaulis Bunge polysaccharide for preventing lung cancer.

[0006] The present application is realized by the following technical scheme:

[0007] The Fritillariae Pallidicaulis Bunge polysaccharide has a polysaccharide content of ≥35%, and in the monosaccharide composition, D-glucose is ≥70%, D-glucuronic acid is ≥3.0%, D-galactose is ≥3.0%, and D-arabinose is ≥3.0%.

[0008] The present application also provides a preparation method of Fritillariae Pallidicaulis Bunge polysaccharide, which comprises the step of extracting Fritillariae Pallidicaulis Bunge.

[0009] Further, the preparation method comprises the step of obtaining crude polysaccharide by defatted water extraction and alcohol precipitation.

[0010] Further, the preparation method further comprises the step of purifying the crude polysaccharide (such as removing protein by macroporous resin).

[0011] In one embodiment of the present application, the preparation method comprises the following steps:

[0012] (1) Pre-treating Fritillariae Thunbergii Miq to obtain defatted Fritillariae Thunbergii Miq powder;

[0013] (2) Extracting the defatted Fritillariae Thunbergii Miq powder with hot water, and then removing protein impurities and small molecules, concentrating, and alcohol precipitation to obtain Fritillariae Thunbergii Miq polysaccharide.

[0014] Further, in step (1), the liquid-to-material ratio of Fritillariae Thunbergii Miq powder to anhydrous ethanol is 1:1-1:10; in one embodiment of the present application, the liquid-to-material ratio is 1:5.

[0015] Further, in step (1), the number of times of extraction is 1-5 times (such as 1, 2, 3, 4, or 5 times); in one embodiment of the present application, the number of times of extraction is 3 times.

[0016] Further, in step (1), the extraction time is 1-4 hours (such as 1, 2, 3, or 4 hours); in one embodiment of the present application, the extraction time is 2 hours.

[0017] In one embodiment of the present application, step (1) comprises: taking Fritillariae Thunbergii Miq powder, adding anhydrous ethanol, boiling point temperature extraction, collecting the precipitate, and obtaining defatted Fritillariae Thunbergii Miq powder.

[0018] Further, in step (2), the liquid-to-material ratio of defatted Fritillariae Thunbergii Miq powder to water is 1:20-50 (such as 1:20, 1:30, 1:40, or 1:50); in one embodiment of the present application, the liquid-to-material ratio is 1:20.

[0019] Further, in step (2), the extraction time is 60-120 min (such as 60 min, 70 min, 80 min, 90 min, or 100 min); in one embodiment of the present application, the extraction time is 120 min.

[0020] Further, in the step of removing protein, macroporous resin is used to remove protein, and in the step of removing impurities and small molecules, membrane separation is used.

[0021] Further, in the step of alcohol precipitation, the alcohol is anhydrous ethanol, and the alcohol content reaches 80%.

[0022] In one embodiment of the present application, step (2) comprises: extracting the defatted Fritillariae Thunbergii Miq powder obtained in step (1) with hot water, collecting the supernatant, removing protein and impurities and small molecules, obtaining Fritillariae Thunbergii Miq polysaccharide solution, concentrating, adding anhydrous ethanol to an alcohol content of 80%, collecting the precipitate after overnight at 4°C, and freeze-drying to obtain Fritillariae Thunbergii Miq polysaccharide.

[0023] The application further provides the crude polysaccharide prepared by the method.

[0024] Further, the lung cancer prevention drug contains the Bulbus Fritillariae Thunbergii polysaccharide extract and pharmaceutically acceptable adjuvants.

[0025] Further, the lung cancer prevention drug is an oral preparation, and the oral preparation is a capsule, a tablet or a granule.

[0026] The application has the following beneficial effects:

[0027] The inventors of the application isolate and purify the Bulbus Fritillariae Thunbergii polysaccharide from the Bulbus Fritillariae Thunbergii, and analyze and identify the molecular weight, monosaccharide composition, infrared spectrum and chemical structure of the Bulbus Fritillariae Thunbergii polysaccharide.

[0028] The application processes the effective components in the Bulbus Fritillariae Thunbergii, and applies the Bulbus Fritillariae Thunbergii polysaccharide extract having the inhibiting effect on the lung cancer cell growth to the preparation of the lung cancer prevention drug, so that the chemical components, the drug efficacy and the action mechanism are combined organically, and a new idea for the fine processing and application of the Bulbus Fritillariae Thunbergii is provided. BRIEF DESCRIPTION OF DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description only the embodiments of the application, and for those skilled in the art, without creative labor, other drawings can also be obtained according to the drawings provided.

[0030] Figure 1 The drawing is the liquid chromatogram of the monosaccharide standard of the Bulbus Fritillariae Thunbergii polysaccharide and the monosaccharide composition analysis of the polysaccharide in experiment 3.

[0031] Figure 2 The drawing is the infrared spectrum of the Bulbus Fritillariae Thunbergii polysaccharide in experiment 3.

[0032] Figure 3 The drawing is the ultraviolet spectrum of the Bulbus Fritillariae Thunbergii polysaccharide in experiment 3.

[0033] Figure 4 The drawing is the liquid phase detection diagram of the determination of the molecular weight of the Bulbus Fritillariae Thunbergii polysaccharide in experiment 3.

[0034] Figure 5 The drawing is the H-NMR detection diagram of the Bulbus Fritillariae Thunbergii polysaccharide in experiment 3. 1 H-NMR detection diagram.

[0035] Figure 6 The drawing is the activity diagram of the Bulbus Fritillariae Thunbergii polysaccharide on A549 and H460 cells in experiment 6.

[0036] Figure 7The figure is a diagram of the effect of the polysaccharide of Fritillariae Thunbergii on the migration and invasion of A549 cells in Experiment 7. DETAILED DESCRIPTION

[0037] The technical solutions of the present application are further described below in connection with examples, but are not limited thereto. Any modification or equivalent replacement of the technical solutions of the present application without departing from the spirit and scope of the technical solutions of the present application shall be covered in the protection scope of the present application.

[0038] Example 1 Extraction and separation and purification of polysaccharide of Fritillariae Thunbergii 1.1 Take a proper amount of dried Fritillariae Thunbergii medicinal materials, crush them by a crushing machine to obtain Fritillariae Thunbergii coarse powder, heat and reflux the Fritillariae Thunbergii coarse powder with 5 times of anhydrous ethanol for 2 hours to remove fat, dry the Fritillariae Thunbergii coarse powder to obtain defatted Fritillariae Thunbergii powder.

[0039] 1.2 Extraction of polysaccharide of Fritillariae Thunbergii Extract the defatted Fritillariae Thunbergii medicinal materials under the conditions of a temperature of 100℃, a liquid-material ratio of 1:20 and an extraction time of 120 minutes, extract for 3 times, combine the filtrates to obtain a polysaccharide of Fritillariae Thunbergii solution.

[0040] 1.3 Removal of impurities and small molecules by hollow fiber membrane Method for separating and purifying the polysaccharide of Fritillariae Thunbergii by hollow fiber membrane: after a certain pretreatment, the polysaccharide of Fritillariae Thunbergii solution is subjected to ultrafiltration by a hollow fiber cellulose membrane with a molecular weight of 5KDa to obtain the polysaccharide of Fritillariae Thunbergii with a molecular weight less than 5KDa and the polysaccharide of Fritillariae Thunbergii with a molecular weight greater than 5KDa; select the part with a molecular weight greater than 5KDa, freeze it to obtain the polysaccharide of Fritillariae Thunbergii solution.

[0041] 1.4 Protein removal by macroporous resin The type of macroporous resin is selected as HP20, and the volume of the pinellia ternate polysaccharide is wet loaded into the macroporous resin HP20 at a certain ratio. The macroporous resin is pretreated. First, soak in water for 18-24 hours, wash with water until the water is clear, pour out the water, and then soak in 4-5% NaOH solution for 24 hours. Wash with water until neutral, add 4-5% HC l solution for 18-24 hours, and then wash with water until neutral. Load the column and load the sample. Take an appropriate amount of pinellia ternate polysaccharide extract solution, add it to the resin chromatographic column after certain treatment, adsorb for a certain time, and then use water as the eluent to start elution at a flow rate of 8-10 ml / min. Track detection by the phenol-sulfuric acid method. When no sugar components flow out, use 0.5-2 mol / L NaCl solution of different concentrations to elute. When no sugar components flow out, use the next gradient of NaCl solution to elute. Concentrate the eluted substances to a small volume, and monitor the density of the concentrated solution with a density meter to obtain a polysaccharide concentrated solution with a density of 1.08-1.15 kg / L. When the concentrated solution is about 50 ℃, add anhydrous ethanol while stirring (to make the alcohol concentration 80%), and then let it stand at 4 ℃ for 24 hours to precipitate the polysaccharide. Wash the precipitate with anhydrous ethanol and acetone, dissolve it in water, and freeze-dry it to obtain crude pinellia ternate polysaccharide.

[0042] Example 2: Determination of the polysaccharide content of pinellia ternate polysaccharide 2.1 Determination of the polysaccharide content Glucose standard solution: accurately weigh 100 mg of glucose standard at 40°C and constant weight, dissolve and dilute to 100 ml in a volumetric flask, and then dilute to 50 ml to prepare a glucose standard solution with a concentration of 0.1 mg / ml. Preparation of the standard curve: accurately measure 0.1 mg / ml glucose standard solution 0, 0.2, 0.4, 0.6, 0.8, 1 ml, respectively, and place it in a 10 ml stoppered test tube. Add double distilled water to 2 ml, and then add 6% phenol reagent 1 ml and shake well. Immediately add 5 ml of concentrated sulfuric acid, shake well, and then place it in a boiling water bath for 15 min. After cooling to room temperature, take out and measure the absorbance value at 490 nm. Prepare a blank control by taking 2.0 ml of distilled water and following the same procedure. Plot the standard curve with the glucose standard solution concentration (μg / mL) as the abscissa and the absorbance value as the ordinate, and calculate the regression equation.

[0043] Sample determination: prepare a sample solution with a concentration of 1 mg / ml. Take 2 ml of the sample solution and follow the above procedure. Measure in triplicate and calculate the polysaccharide content according to the standard curve regression equation.

[0044] 2.2 Detection results: the standard regression equation is y = 0.0021x + 0.1199 R² = 0.9951, and the content of pinellia ternate polysaccharide is ≥ 35%.

[0045] Structure identification of the polysaccharide from the bulb of Fritillaria ussuriensis Maxim. 3.1 Analysis of monosaccharide composition The monosaccharide standards (D-glucose, D-mannose, D-glucuronic acid, D-rhamnose, D-galactose, D-galacturonic acid, D-arabinose) and 10.00 mg of the polysaccharide from the bulb of Fritillaria ussuriensis Maxim. after complete acid hydrolysis (trifluoroacetic acid method) were analyzed by high performance liquid chromatography (HPLC) using NaH2PO4 and acetonitrile (82:18) as the mobile phase at a flow rate of 1 ml / min.

[0046] 3.2 Infrared spectrum analysis The polysaccharide from the bulb of Fritillaria ussuriensis Maxim. (3 mg) was ground with a certain amount of dry KBr in a mortar and pressed into a tablet using a tablet press. The sample spectrum was obtained by Fourier transform of the obtained tablet, with a range of 4000-400 cm -1 -1 and a resolution of 4 cm -1 -1. Background collection (KBr) was required before collecting the sample. The obtained sample graph was automatically baseline calibrated and automatically smoothed.

[0047] 3.3 Ultraviolet spectrum analysis The polysaccharide from the bulb of Fritillaria ussuriensis Maxim. 5 mg was dissolved in a 10 ml volumetric flask to prepare a 0.5 mg / mL solution. After centrifugation, the supernatant was scanned at 200-400 nm using an ultraviolet spectrophotometer to determine whether the polysaccharide contained nucleic acid and protein conjugates.

[0048] 3.4 Determination of molecular weight 10 mg of the polysaccharide from the bulb of Fritillaria ussuriensis Maxim. (obtained in Example 1) was accurately weighed, dissolved in 1 mL of H2O, and ultrasonically treated for 5 min. Then 100 μL of the solution was dissolved in 900 μL of water to prepare a 1 mg / ml sample, which was filtered with a 0.22 μm filter membrane and ready for gel permeation chromatography analysis.

[0049] 3.5 Nuclear magnetic resonance analysis 20 mg of the polysaccharide from the bulb of Fritillaria ussuriensis Maxim. was weighed and dissolved in 5.5 ml of D2O. After centrifugation, the supernatant was taken and placed in a nuclear magnetic tube. The determination of H-NMR spectrum was performed on a nuclear magnetic resonance spectrometer. 1

[0050] 3.6 Detection results 3.6.1 Analysis of monosaccharide composition of the polysaccharide from the bulb of Fritillaria ussuriensis Maxim. After hydrolysis of the polysaccharide from the bulb of Fritillaria ussuriensis Maxim. into monosaccharides, high performance liquid chromatography was used for monosaccharide composition analysis, and the results are shown in Table 1. Figure 1 ​As shown. Its main monosaccharide component is glucose, with a retention time of 33.513 min. Glucuronic acid, galactose, and arabinose are minor monosaccharide components.

[0051] 3.6.2 Infrared spectral analysis results of Fritillaria cirrhosa polysaccharides Fourier transform infrared spectrum of Fritillaria cirrhosa polysaccharide as follows Figure 2 As shown, at 3310.61cm -1 At almost all locations, a broad and large absorption peak exists, which is the stretching vibration absorption band of OH; at 2920.45 cm⁻¹... -1 A weak absorption peak is observed at 1655.30 cm⁻¹, indicating the stretching vibration of the methylene group; this is a characteristic peak of polysaccharides. -1 and 1420.45 cm -1 It originates from the carboxylic acid anionic bond (C=O), and is located at 928.03.58 cm⁻¹. -1 An absorption band at the carboxyl group can be observed, confirming the presence of uronic acid in Fritillaria cirrhosa polysaccharide. (1155.30 cm) -1 1087.12cm -1 A set of peaks represents the vibrational absorption bands of COC and CO in the sample, indicating the presence of the pyran ring, also at 757.58 cm⁻¹. -1 This further confirms the presence of a pyran ring in the sample. 852.27cm -1 The characteristic peak at that location indicates the presence of an α-glycosidic bond.

[0052] 3.6.3 Ultraviolet Spectroscopic Analysis of Fritillaria cirrhosa Polysaccharides Depend on Figure 3 The results showed that the crude polysaccharide of Fritillaria cirrhosa had an absorption peak at 260-280 nm, but the purified Fritillaria cirrhosa polysaccharide did not have an obvious absorption peak at 260-280 nm, indicating that the Fritillaria cirrhosa polysaccharide did not contain nucleic acids, proteins and other substances, and the experimental method had basically removed such substances.

[0053] 3.6.4 Determination of the molecular weight of Fritillaria cirrhosa polysaccharide The molecular weight of Fritillaria cirrhosa polysaccharide is as follows Figure 4 As shown, a standard curve was constructed based on molecular weight and retention time, with the following equation: y = -0.2654x + 7.5665, R² = 0.9911. The graph indicates that the molecular weight range of Fritillaria cirrhosa polysaccharide is 2 × 10⁻⁶. 2 KDa ~1.5×10 3 KDa.

[0054] 3.6.5 NMR analysis results of Fritillaria cirrhosa polysaccharides 1The H-NMR spectrum shows that the signals of polysaccharide are mostly concentrated in the range of δ 3.1-5.4. The signals of δ 3.1-4.2 belong to the protons on the sugar ring carbons C2-C6. The spectrum shows that in the range of δ 4.5-5.5, six end group hydrogen signals of δ 4.91, 4.98, 5.10, 5.17, 5.24 and 5.40 can be observed, and in combination with the HPLC monosaccharide composition results, it is preliminarily judged that Fritillariae Pallidiflorae Bulbus contains at least four kinds of monosaccharides. The signal intensity of the protons with a chemical shift greater than δ 5.0 is obviously stronger than that of the protons with a chemical shift less than δ 5.0, indicating that the glycosidic configuration of Fritillariae Pallidiflorae Bulbus polysaccharide is mainly of the α-type, which is consistent with the results of IR spectrum analysis. δ 4.91 is β-D-GlcpA, δ 4.98 is α-D-Galp, δ 5.17 is α-D-Galp, δ 5.24 is α-D-Araf, and δ 5.40 is α-D-Glcp.

[0055] Example 4 Activity research of Fritillariae Pallidiflorae Bulbus polysaccharide extract in preventing lung cancer 4.1 Experimental materials: 72 C57BL / 6 male mice, Fritillariae Pallidiflorae Bulbus polysaccharide obtained by the preparation method provided in Example 1 4.2 Experimental method: Animal grouping: The mice were placed in a condition of relative humidity of 65% and temperature of 22 ℃ for one week of systematic culture, and the mice were randomly grouped: ① blank group; ② model group; ③ low-dose Fritillariae Pallidiflorae Bulbus group; and ④ high-dose Fritillariae Pallidiflorae Bulbus group. The administration mode was as follows: the blank group was given normal saline 200 uL / mouse / day until the end of the experiment; the model group was intraperitoneally injected with urethane 800 mg / kg / time, once a week, for 8 weeks; the low-dose Fritillariae Pallidiflorae Bulbus group was given Fritillariae Pallidiflorae Bulbus polysaccharide solution 0.45 g / kg / mouse / day by gavage, and the high-dose Fritillariae Pallidiflorae Bulbus group was given Fritillariae Pallidiflorae Bulbus polysaccharide solution 1.35 g / kg / mouse / day by gavage until the end of the experiment. At the end of the experiment, the number and size of tumors were detected, the body weight of the mice was weighed, the autonomous activity ability was measured, and the morphological changes of the lung tissue of the mice were observed.

[0056] 4.3 Experimental results: The tumor volume was recorded once a day, and the tumor volume estimation formula was: tumor volume V (cm 3 ) = 1 / 2 × a × b 2 The high-dose and low-dose Fritillariae Pallidiflorae Bulbus polysaccharide extract can improve the body weight of the urethane-induced lung cancer model mice, improve the survival quality of the mice, increase the number of autonomous activities, and improve the pathological changes of lung cancer.

[0057] The tumor inhibition rate calculation formula was: (1-experimental group volume / blank group volume) × 100% Group Administration mode Initial body weight (g) Initial number of cases (only) Final body weight (g) Final number of cases (only) Tumor inhibition rate (%) Blank group Normal saline 23.2±0.5 10 26.5±0.8 10 100 Model group Normal saline 23.3±0.6 10 20.6±0.5 6 0 Low-dose group 0.45 g / kg 22.9±0.4 10 22.1±0.7 7 28.57 High-dose group 1.35 g / kg 23.6±0.7 10 23.5±0.6 8 75.00 Positive drug group Aspirin 22.8±0.6 10 23.0±0.8 8 62.5 4.4 Data processing SPSS 26 software was used to analyze the significant differences between groups by one-way ANOVA combined with the least significant difference method. All the data shown are mean ± SD.

[0058] Example 5 Inhibitory effect of polysaccharides from Fritillaria ussuriensis on the proliferation of two lung cancer cells 5.1 Experimental materials: Human large cell lung cancer cells H460 and human lung cancer cell line A549 were purchased from the Chinese Academy of Sciences Cell Bank. Polysaccharides from Fritillaria ussuriensis were obtained in Example 1.

[0059] 5.2 Experimental method: A549 lung cancer cells in good growth condition were taken, and 5x10 3 cells / ml of cells were inoculated in a 96-well cell culture plate, 6 replicates were set for each group. After the cells adhered, the culture medium was discarded, 200 μl of culture solution was added to the blank control group, and different concentrations of polysaccharide solution from Fritillaria ussuriensis were added to the experimental groups. After incubation in the cell culture box for 24 hours, 20 μl of MTT solution was added, and incubation was continued for 4 hours. Then 100 μl of dimethyl sulfoxide was added, and the shaking bed was shaken for 10 minutes at room temperature. The absorbance value of each well was measured at a wavelength of 490 nm.

[0060] 5.3 Experimental results: The results of the activity of crude polysaccharides from Fritillaria ussuriensis on A549 and H460 cells are shown in the accompanying Figure 6 , a is A549, b is H460. The MTT experiment results show that after different concentrations of polysaccharides from Fritillaria ussuriensis were applied to human lung cancer cells A549 and H460, cell proliferation was inhibited, and this inhibition was dose-dependent. With the continuous increase of the concentration of polysaccharides from Fritillaria ussuriensis, the inhibition of A549 cell proliferation was more obvious. There were differences between different concentrations, and in the concentration range of 125-500 μg / mL, the inhibition rate increased continuously with the increase of the concentration of polysaccharides from Fritillaria ussuriensis (p<0.05). It was found that polysaccharides from Fritillaria ussuriensis had significant inhibitory activity on lung cancer cells.

[0061] Example 6 Effect of polysaccharides from Fritillaria ussuriensis on A549 cell migration and invasion

[0062] 6.1 Experimental materials: Human lung cancer cell line A549, polysaccharides from Fritillaria ussuriensis.

[0063] 6.2 Experimental method: The effect of polysaccharides from Fritillaria ussuriensis on A549 cell migration and invasion was qualitatively observed by cell scratch test. 1.5x10 6After 24h, the cells were observed under microscope and confirmed that the cells had covered the 6-well plate. After the scratch was drawn, the cells were washed with PBS for 3 times. The cells were intervened with different concentrations of the polysaccharides of Fritillariae ussuriensis while setting up a control group. The cells were placed in a 37℃ incubator containing 5% CO2 and continued to be cultured. The samples were taken at 0h, 24h and 48h for photographing.

[0064] 6.3 Experimental results: The changes of the scratch area of A549 cells within 48h were observed under an inverted microscope. The results are shown in FIG. 6. Figure 7 The results show that the number of cells migrating in the scratch damage area of the intervention group of the polysaccharides of Fritillariae ussuriensis is significantly less than that of the blank control group, and the area of the scratch area covered by cells is not obvious. The scratch damage area of the blank control group is covered by lung cancer cells again, which indicates that the polysaccharides of Fritillariae ussuriensis have a trend of inhibiting the migration of A549 cells.

[0065] The embodiments in the specification are described in a progressive manner, and each embodiment focuses on the difference from other embodiments. The same or similar parts between the embodiments can be referred to each other. The general principles defined in this document can be implemented in other embodiments without departing from the spirit or scope of the present application. Therefore, the present application will not be limited to the embodiments shown in this document, but will conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A polysaccharide from Fritillaria cirrhosa, characterized in that, The Fritillaria cirrhosa polysaccharide is composed of glucuronic acid, glucose, galactose, and arabinose. After hydrolysis, the proportions are: D-glucose ≥ 70%, D-glucuronic acid ≥ 3.0%, D-galactose ≥ 3.0%, and D-arabinose ≥ 3.0%. The molecular weight range of the Fritillaria cirrhosa polysaccharide is 2 × 10⁻⁶. 2 Kda ~1.5×10 3 KDa.

2. The method for preparing the polysaccharide according to claim 1, wherein the polysaccharide is obtained by extracting it from the bulb of Fritillaria cirrhosa.

3. A method for preparing Fritillaria cirrhosa polysaccharide as described in claim 2, characterized in that, Includes the following steps: Step 1: Extraction of Fritillaria cirrhosa polysaccharides: Fritillaria cirrhosa is pulverized, sieved, defatted with ethanol, filtered, and dried to obtain a solid medicinal powder. The solid medicinal powder is then extracted multiple times with water, filtered, and the filtrates are combined to obtain a crude Fritillaria cirrhosa polysaccharide solution. Step 2: Removal of protein and small impurities from the crude Fritillaria cirrhosa polysaccharide: The crude Fritillaria cirrhosa polysaccharide solution obtained in Step 1 is subjected to membrane separation and macroporous resin method to remove protein and small impurities. The eluent is collected, concentrated by rotary evaporation, and precipitated with alcohol to obtain Fritillaria cirrhosa polysaccharides.

4. The use of the polysaccharide and its preparations as described in claim 2 in the preparation of drugs for treating and preventing cancer.

5. The use of the polysaccharide and its preparations as described in claim 2 in the preparation of drugs for the treatment and prevention of lung cancer.

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