Extraction method of bulbophyllum polysaccharide SDLN1 and application thereof

Abstract

The application relates to an extraction method of stone orchid polysaccharide SDLN1 and application thereof, and can effectively solve the medicinal problem of the stone orchid polysaccharide. The technical scheme is that the structure of the stone orchid polysaccharide SDLN1 is as shown in the following formula: a novel structure of the stone orchid polysaccharide is extracted, and further research shows that the stone orchid polysaccharide extracted by the application has a certain protective effect on the lung tissue of an LPS-induced ALI model mouse, can be used as an active ingredient of an anti-inflammatory drug, and is applied to the preparation of the anti-inflammatory drug, and is an innovation in the extraction method of the stone orchid polysaccharide.

Description

I. TECHNICAL FIELD

[0001] The present application relates to the field of medicine extraction, in particular to an extraction method of a Bulbophyllum polysaccharide SDLN1 and application thereof. II. BACKGROUND

[0002] Bulbophyllum is a kind of Chinese herbal medicine commonly used in China, which has the effects of antibacterial, anti-inflammatory, antioxidant and anticholinesterase activity. Polysaccharide is one of the main active components of Bulbophyllum. At present, the research on Bulbophyllum mainly focuses on resources, identification or small molecule compounds, and the research on polysaccharide is less. At present, there is no research on the extraction and preparation method of Bulbophyllum polysaccharide, and the separation and purification, structure analysis of Bulbophyllum polysaccharide need further research.

[0003] Acute lung injury (ALI) is a common reaction to various infectious and non-infectious injuries, which can be caused by inflammatory stress, and can cause acute respiratory distress syndrome and multiple organ failure, which is life-threatening. Polysaccharide is a natural high molecular compound, which generally exists in animals and plants, mainly distributed in animal cell membranes and plant cell walls. Pharmacological studies have shown that polysaccharide has antiviral, anti-inflammatory, antioxidant, antitumor and other biological activities, and therefore has attracted more and more attention of researchers, especially in the aspect of anti-inflammatory. Bulbophyllum can nourish yin, clear heat and reduce fire, and is commonly used in folk to treat sore throat, pneumonia, tuberculosis and the like. At present, the main active ingredient of Bulbophyllum for anti-inflammatory is not clear, and whether Bulbophyllum polysaccharide is the main active ingredient has not been reported. Therefore, it is necessary to analyze the structure and application of Bulbophyllum polysaccharide, so as to obtain Bulbophyllum polysaccharide with anti-inflammatory effect. III. SUMMARY

[0004] In view of the above situation, in order to solve the defects of the prior art, the purpose of the present application is to provide an extraction method of Bulbophyllum polysaccharide SDLN1 and application thereof, which can effectively solve the medicinal problems of Bulbophyllum polysaccharide.

[0005] The technical solution solved by the present application is that the structure of the Bulbophyllum polysaccharide SDLN1 is as follows:

[0006]

[0007] The extraction method of the Bulbophyllum polysaccharide comprises the following steps:

[0008] 1) Extracting defatted Bulbophyllum

[0009] The dry Bulbophyllum sample is crushed and subjected to petroleum ether defatting treatment to obtain defatted Bulbophyllum;

[0010] 2) Extracting Bulbophyllum polysaccharide crude product

[0011] The defatted Anoectochilus roxburghii extracted in step 1) is added into water, and then heated to carry out reflux extraction; after the extraction is completed, the extraction liquid is concentrated to obtain a concentrated liquid, and ethanol is added to the concentrated liquid to carry out alcohol precipitation, so as to obtain an Anoectochilus roxburghii polysaccharide crude product;

[0012] 3) Extraction of Anoectochilus roxburghii refined crude polysaccharide

[0013] The Anoectochilus roxburghii polysaccharide crude product extracted in step 2) is redissolved in water, and then subjected to Sevag method deproteinization, decolorization and dialysis treatment, so as to obtain Anoectochilus roxburghii refined crude polysaccharide.

[0014] 4) Purification of Anoectochilus roxburghii refined crude polysaccharide

[0015] The Anoectochilus roxburghii refined crude polysaccharide extracted in step 3) is subjected to separation and purification treatment of DEAE-52 cellulose column and Sephadex G-100 column in sequence, and then freeze-dried, so as to obtain Anoectochilus roxburghii polysaccharide SDLN1.

[0016] The Anoectochilus roxburghii polysaccharide SDLN1 is applied to the preparation of an anti-inflammatory drug.

[0017] The Anoectochilus roxburghii polysaccharide extracted in the application has a novel structure, and further research shows that the Anoectochilus roxburghii polysaccharide extracted in the application has a certain protective effect on the lung tissue of an LPS-induced ALI model mouse, can be used as an active ingredient of an anti-inflammatory drug, and can be applied to the preparation of an anti-inflammatory drug, which is an innovation in the extraction method of Anoectochilus roxburghii polysaccharide. IV. DESCRIPTION OF DRAWINGS

[0018] Figure 1 It is an elution curve diagram of the 0.1 Nacl part of the Anoectochilus roxburghii polysaccharide in the application.

[0019] Figure 2 It is an infrared characteristic diagram of the SDLN1 extracted in the application.

[0020] Figure 3 It is an ion chromatogram of a standard product in the application.

[0021] Figure 4 It is an ion chromatogram of a sample in the application.

[0022] Figure 5 It is 1H NMR spectrum (A), 13C nuclear magnetic resonance spectrum (B), HSQC spectrum (C), COSY spectrum (D), HMBC spectrum (E) and NOESY spectrum (F) of the SDLN1 in the application.

[0023] Figure 6 It is the influence of the SDLN1 in the application on the total protein in the lung alveolar lavage fluid, the lung tissue and the TNF-α and IL-6 levels in the lung alveolar lavage fluid of each group of mice; note: compared with the blank group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ## P<0.01.

[0024] Figure 7 The pathological changes of lung tissue in mice in each group of the present invention (HE staining, A: blank group, B: model group, C: DEX group, D: SDLN1-L group, E: SDLN1-H group). V. Detailed Implementation Methods

[0025] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings and examples.

[0026] Example 1

[0027] In its specific implementation, this invention includes the following steps:

[0028] 1) Extraction of defatted Bulbus orchid

[0029] Take 2 kg of dried Bulbus burgdorferi, crush it into powder, add petroleum ether at a material-to-liquid ratio of 1:20, reflux extract at 90℃ for 1 h, filter to obtain defatted Bulbus burgdorferi.

[0030] 2) Extraction of crude polysaccharide from *Bulbus buergerianum*

[0031] The defatted Bryophyllum filter residue extracted in step 1) was dried, and distilled water was added at a material-to-liquid ratio of 1:20. The mixture was extracted three times by hot water reflux, each time for 1 hour, and then filtered. The filtrate was concentrated under reduced pressure to 2000 mL, and ethanol was added to make the alcohol content reach 80%. The mixture was allowed to stand overnight, and the precipitate was collected by centrifugation and then freeze-dried under vacuum to obtain crude Bryophyllum polysaccharide.

[0032] 3) Extraction of refined crude polysaccharides from *Bulbus pachymansis*

[0033] The crude polysaccharide extracted in step 2) was dissolved in distilled water and deproteinized using the Sevag method (chloroform: n-butanol = 4:1). The volume ratio of polysaccharide solution to Sevag reagent was 3:1. Then, the pigment was removed by H2O2 method, small molecules were removed by dialysis, the solution was collected, and vacuum freeze-dried to obtain refined crude polysaccharide from Dendrobium nobile.

[0034] 4) Purification of crude polysaccharides from *Bulbus pachycarpa*

[0035] 100mg of the purified crude polysaccharide extracted in step 3) is dissolved in 10ml of water and added to an equilibrated DEAE-52 cellulose column, and eluted with a NaCl gradient solution (0-0.5Mol / L) in sequence, and different gradient Burmannia polysaccharide eluate is collected, concentrated, dialyzed and freeze-dried to obtain different elution components of the polysaccharide; the polysaccharide components are tracked by the phenol sulfuric acid method, the peaks containing the polysaccharide are collected and freeze-dried, and according to the results of each elution component, the 0.1 NaCl elution part is selected for further purification by a gel purification system, the polysaccharide components are collected, concentrated, dialyzed and freeze-dried to obtain Burmannia polysaccharide SDLN1.

[0036] The extraction method of the present application extracts a brand new structure of Burmannia polysaccharide, and research shows that Burmannia polysaccharide SDLN1 can reduce the inflammatory response of LPS-induced ALI model mice, has a certain protective effect on the lung tissue of LPS-induced ALI model mice, and can be applied to anti-inflammatory drugs, and the related test data are as follows:

[0037] I. Structure characterization of Burmannia polysaccharide SDLN1

[0038] 1.1 Molecular weight determination

[0039] The gel chromatography-differential-multiple angle laser light scattering system is used to determine the molecular weight of the polysaccharide, the liquid phase system is U3000 (Thermo, USA), the differential detector is Optilab T-rEX (Wyatt technology, CA, USA), and the laser light scattering detector is DAWN HELEOS II (Wyatt technology, CA, USA). The gel exclusion chromatography column Ohpak SB-805HQ (300*8mm) and Ohpak SB-803HQ (300*8mm) are connected in series. The column temperature is 45℃, the sample injection amount is 100μL, the flow phase A (0.02% NaN3, 0.1M NaNO3) is used, the flow rate is 0.6mL / min, and the elution gradient is isocratic for 75min. The chromatography data is processed by software ASTRA6.1. The results show that the absolute molecular weight of the polysaccharide is 39.937KDa.

[0040] 1.2 Fourier transform infrared spectroscopy (FT-IR) analysis

[0041] The dried SDLN1 is combined with KBr at a ratio of 1:100 to obtain transparent tablets, and then scanned by FT-IR (Nicolet 6700, Thermo) in the range of 4000-400cm-1, and the infrared spectrum is as follows: Figure 2As shown in the figure, a strong and broad absorption peak exists at 3412.89 cm⁻¹, which is a strong absorption peak of the OH stretching vibration of hydrogen bonds between or within polysaccharide molecules, and is a characteristic peak of carbohydrates. The peak at 2934.85 cm⁻¹ is the absorption peak of CH stretching vibration, and there is an absorption peak at 1637 cm⁻¹, which may be attributed to the C=O asymmetric stretching vibration. There are absorption peaks at 1402 m⁻¹ and 1050 cm⁻¹, which may be attributed to the CO stretching vibration.

[0042] 1.3 Monosaccharide Composition Analysis

[0043] SDLN1 sample was hydrolyzed in 1 ml of 2M TFA acid solution at 121℃ for 2 h, dried under nitrogen, rinsed with methanol, dried under nitrogen, and repeated 2-3 times. The solution was then dissolved in distilled water and transferred to a chromatographic vial for analysis. The monosaccharide components were analyzed and detected using a Thermo ICS 5000+ ion chromatography system (ICS 5000+, Thermo Fisher Scientific, USA). Specific chromatographic conditions were as follows: Dionex TM CarboPac TM PA20 (150*3.0mm, 10μm) liquid chromatography column; injection volume: 5μl. Mobile phase A (H2O), mobile phase B (0.1M NaOH), mobile phase C (0.1M NaOH, 0.2M NaAc), flow rate 0.5 ml / min; column temperature 30℃; elution gradient: 0 min A phase / B phase / C phase (95:5:0, V / V), 26 min A phase / B phase / C phase (85:5:10, V / V), 42 min A phase / B phase / C phase (85:5:10, V / V), 42.1 min A phase / B phase / C phase (60:0:40, V / V), 52 min A phase / B phase / C phase (60:40:0, V / V), 52.1 min A phase / B phase / C phase (95:5:0, V / V), 60 min A phase / B phase / C phase (95:5:0, V / V). The results showed that SDLN1 was mainly composed of fructose, rhamnose, arabinose, galactose, glucose, xylose, and mannose, with a molar ratio of 4.66:10.18:9.96:27.86:15.32:10.56:19.49:1.97. The chromatograms of the standard and samples are shown below. Figures 3-4 As shown.

[0044] 1.4 Methylation Analysis

[0045] A small amount of SDLN1 sample (2-3 mg) was dissolved in 500 μΐ DMSO. 1 mg NaOH was added and incubated for 30 min, then 50 μΐ iodomethane solution was added and reacted for 1 h. After the reaction, 1 ml water and 2 ml dichloromethane were added, vortexed, centrifuged, and the water phase was discarded. This was repeated three times. Then the lower dichloromethane phase was taken and blown dry with nitrogen, followed by the addition of 100 μΐ 2M TFA, reacted at 121 °C for 90 min, and evaporated at 30 °C. Subsequently, 50 μΐ 2M ammonia water and 50 μΐ 1M NaBD4 were added, mixed, reacted at room temperature for 2.5 h, terminated with 20 μΐ acetic acid, blown dry with nitrogen, washed twice with 250 μΐ methanol, and blown dry with nitrogen. Then 250 μΐ acetic anhydride was added, vortexed, reacted at 100 °C for 2.5 h, and 1 ml water was added and allowed to stand for 10 min. Subsequently, 500 μΐ dichloromethane was added, vortexed, centrifuged, and the water phase was discarded, and the water washing was repeated three times. The lower dichloromethane phase was taken and subjected to GC-MS analysis. The analysis was performed using an Agilent gas chromatograph-mass spectrometer. The chromatographic conditions were as follows: chromatographic column: BPX70 (30 m x 0.25 mm x 0.25 μιη, SGE, Australia). The injection volume was 1 μΐ, the split ratio was 10:1, the carrier gas was high-purity helium, the flow rate was 1.5 ml / min, and the initial temperature of the column oven was 140 °C, maintained for 2.0 min, then programmed to increase at 3 °C / min to 230 °C, maintained for 3 min. The electron impact ion source (EI) and the mass scan range (m / z) were 50-350. The results showed that SDLN1 contained 18 different types of bonds, with the largest proportion of galactose residues, and the presence of t-Gal(p), 3-Gal(p), 4-Gal(p), 2,4-Gal(p) and 3,6-Gal(p) sugar bonds, followed by mannose residues and glucose residues, by mass spectrometric identification of the ion fragments (as shown in Table 1).

[0046] Table 1. Results of methylation analysis of SDLN1

[0047]

[0048]

[0049] 1.5 NMR analysis

[0050] A 50 mg sample of SDLN1 was dissolved in 0.5 mL D2O, freeze-dried, and this process was repeated twice. Then the sample was dissolved in 0.5 mL D2O, and 1H-NMR, 13C-NMR, chemical shift correlation (COSY), heteronuclear single quantum correlation (HSQC), two-dimensional NOE spectrum (NOESY), and heteronuclear multiple carbon correlation (HMBC) were obtained using a Bruker 600 nuclear magnetic resonance instrument (Billerica, MA, USA) (as shown in Figure 5 Table 2. NMR analysis of SDLN1

[0051] As can be seen from the figure, the sample hydrogen spectrum signal is mainly concentrated between δ3.0-5.5ppm, and multiple coupling signal peaks are identified in the anomeric signal region of δ4.3-5.4ppm, indicating that the sample contains multiple sugar residues, and the chemical shifts of the anomeric hydrogens are δ4.41, 4.5, 4.61, 4.62, 4.95, 5.06, 5.21, 5.39ppm, etc. The non-anomeric hydrogen signal is mainly concentrated in the δ3.1-4.2ppm region, and individual signals need to be assigned to the H2-H6 chemical shifts of each sugar residue due to severe overlap. The strong signal peak near δ4.71ppm is the solvent peak. The signal peak near δ3.16ppm is the signal of the hydrogen of O-CH3.

[0052] The sample recognizes multiple signal peaks in the anomeric carbon region, and the cross peaks in the anomeric region of 13C NMR and HSQC spectra determine that the anomeric signals present in the sample are: δ5.06 / 108.87, 4.41 / 103.29, 4.5 / 101.85, 4.61 / 103.55, 5.39 / 97.21, 4.62 / 101.02, 5.21 / 97.64, 4.95 / 99.03ppm, and are respectively denoted as sugar residues A, B, C, D, E, F, G, I. Combined with the sample bonding structure (methylation) information, anomeric signals and comprehensive reports in the literature, it is speculated that sugar residue A is α-L-Araf-(1→, sugar residue B is →4)-β-D-Galp-(1→, sugar residue C is →3,4)-β-D-Glcp-6-OMe-(1→, sugar residue D is →3)-β-D-Glcp-(1→, sugar residue E is →3)-α-D-Galp-(1→[8], sugar residue F is →4)-β-D-Manp-(1→, sugar residue G is →2)-α-D-Manp-(1→, sugar residue I is →3,6)-α-D-Galp-(1→. The 1H and 13C chemical shifts are assigned, and the results are shown in Table 2.

[0053] Table 2 Sugar residues 1 H and 13 C chemical shifts

[0054]

[0055] n.d: The abbreviation of "not detected" means not identified.

[0056] The structure and linkage of the polysaccharide were analyzed according to the chemical shifts of 13C and 1H of each sugar residue in the sample, combined with HMBC and NOESY spectrum analysis. According to the HMBC spectrum, there is a cross peak between H1 of sugar residue A and C4 of sugar residue C δ5.06 / 82.9ppm. According to the NOESY spectrum, there is a cross peak between H1 of sugar residue A and H4 of sugar residue C δ5.06 / 4.23ppm, a cross peak between H1 of sugar residue A and H2 of sugar residue G δ5.06 / 3.5ppm, a cross peak between H1 of sugar residue B and H4 of sugar residue B δ4.41 / 3.38ppm, a cross peak between H1 of sugar residue B and H4 of sugar residue F δ4.41 / 4.06ppm, a cross peak between H1 of sugar residue C and H3 of sugar residue I δ4.5 / 4.09ppm, a cross peak between H1 of sugar residue D and H3 of sugar residue E δ4.61 / 3.64ppm, a cross peak between H1 of sugar residue E and H4 of sugar residue B δ5.39 / 3.38ppm, a cross peak between H1 of sugar residue F and H3 of sugar residue C δ4.62 / 3.89ppm, a cross peak between H1 of sugar residue G and H6 of sugar residue I δ5.21 / 3.95ppm, a cross peak between H1 of sugar residue G and H6 of sugar residue I δ5.21 / 4.01ppm, and a cross peak between H1 of sugar residue I and H3 of sugar residue D δ4.95 / 3.73ppm. Therefore, according to the analysis of one-dimensional nuclear magnetic and two-dimensional nuclear magnetic information and methylation results, it is inferred that the polysaccharide is mainly connected by →4)-β-D-Galp-(1→, →3,4)-β-D-Glcp-6-OMe-(1→ and →3)-β-D-Glcp-(1→ to form the main chain, and the branched chain is mainly composed of α-L-Araf-(1→ connected to O-4 of sugar residue →3,4)-β-D-Glcp-6-OMe-(1→, etc. Therefore, the possible structure of the polysaccharide chain is as follows:

[0057]

[0058] II. Pharmacological effects of Anoectochilus roxburghii polysaccharide SDLN1 on acute lung injury (ALI)

[0059] 2.1 Method

[0060] 2.1.1 Establishment of ALI mouse model and administration

[0061] 40 SPF level 6-week-old male C57BL / 6J mice were adaptively fed for 7 days, and then the mice were randomly divided into a blank group, a model group, a dexamethasone group (DEX group), a low-dose SDLN1 administration group (SDLN1-L group, 20 mg / kg / d), and a high-dose SDLN1 administration group (SDLN1-H group, 40 mg / kg / d), with 8 mice in each group. Before modeling, the DEX group was intraperitoneally injected with 10 mg / kg of DEX, and the SDLN1-H group and the SDLN1-L group were given SDLN1 treatment for 3 days before modeling. Subsequently, ALI modeling was performed by intratracheal instillation of LPS, that is, ALI was induced by intratracheal administration of Escherichia coli 055:B5 lipopolysaccharide, and 24 hours after LPS infusion, the mice were sacrificed, and the left lung was lavaged twice with PBS to obtain the bronchoalveolar lavage fluid (BALF). At the same time, the right lung tissue was divided into two parts, one of which was immersed in a 10% formalin solution for histopathological examination, and the other was used for determination of inflammatory factors.

[0062] 2.1.2 Detection of total protein in the bronchoalveolar lavage fluid by BCA method

[0063] The bronchoalveolar lavage fluid was taken, and the total protein concentration was determined using a BCA protein concentration detection kit, according to the instructions of the kit.

[0064] 2.1.3 Detection of TNF-α and IL-6 levels by enzyme-linked immunosorbent assay (ELISA)

[0065] The concentrations of TNF-α and IL-6 in the BALF and lung tissue extract supernatant were determined using a kit. The supernatant was obtained by centrifugation of the BALF and was ready for use. The lung tissue was weighed, then homogenized with PBS, centrifuged at 12,000g at 4°C for 15 minutes, and the supernatant was taken and ready for use.

[0066] 2.1.4 Histopathological changes in lung tissue

[0067] The right lung tissue of the mice was fixed with 10% formalin, paraffin-embedded, sectioned, and stained with hematoxylin-eosin (HE), and histopathological observation was performed under a microscope.

[0068] 2.1.5 Statistical methods

[0069] SPSS 22.0 software and Graphpad Prism 8.0 software were used to analyze the experimental data. The measurement data were expressed as mean ± standard deviation (xˉ±s). The t-test was used for comparison between two groups, and the one-way analysis of variance was used for comparison among multiple groups. P<0.05 was considered statistically significant.

[0070] 2.2 Results

[0071] 2.2.1 Levels of total protein in BALF, TNF-α and IL-6 in lung tissue and BALF

[0072] Compared with the control group, the levels of BCA, TNF-α and IL-6 in BALF of the LPS group were significantly increased (p<0.05), and the levels of TNF-α and IL-6 in lung homogenate tissue were significantly increased (p<0.05), indicating that the lung of the model mice had a certain inflammatory response. After treatment with the Neolitsea polysaccharide SDLN1, the levels of inflammatory factors decreased significantly, and the levels of BCA, IL-6 in lung homogenate tissue, TNF and IL-6 in BALF of the SDLN1-H group were significantly reduced (p<0.05). The level of TNF in BALF of the SDLN1-L group was significantly reduced (p<0.05), and the levels of TNF, IL-6 in lung homogenate tissue and IL-6 in BALF had a decreasing trend, indicating that the Neolitsea polysaccharide SDLN1 could alleviate the inflammatory response of the LPS-induced ALI model mice, as shown in Figure 6 .

[0073] 2.2.2 Pathological observation of lung tissue of mice

[0074] The lung tissue structure of the blank group mice was complete, and no obvious pathological changes were observed. Compared with the blank group, the alveolar epithelial cells of the model group mice proliferated, the alveolar septum was significantly thickened, and a large number of inflammatory cells were observed in the interstitium. The low-dose Neolitsea polysaccharide group showed a certain degree of improvement, and part of the alveolar epithelial cells proliferated, the alveolar septum was thickened, and more inflammatory cells were observed in the interstitium. The low-dose and high-dose Neolitsea polysaccharide groups had a more obvious improvement effect, and the inflammatory cells in the lung tissue were significantly reduced, indicating that the Neolitsea polysaccharide had a certain protective effect on the lung tissue of the LPS-induced ALI model mice, as shown in Figure 7 .

[0075] III. CONCLUSION

[0076] In summary, the Neolitsea polysaccharide SDLN1 with a novel structure is extracted, and research shows that the Neolitsea polysaccharide SDLN1 can alleviate the inflammatory response of the LPS-induced ALI model mice, has a certain protective effect on the lung tissue of the LPS-induced ALI model mice, and can be applied to the production and development of anti-inflammatory drugs. The new structure and new use of the Neolitsea polysaccharide are developed, which has practical clinical application and popularization value, and has huge economic and social benefits.

[0077] It should be pointed out that the above is only a preferred embodiment of the present application, and does not limit the present application in any form. Any person skilled in the art can make changes or modifications to the equivalent embodiments within the scope of the technical solutions of the present application by using the disclosed technical content, which falls within the protection scope of the present application.

Claims

1. A Bulbophyllum polysaccharide SDLN1, characterized in that, The structure of the Bulbophyllum polysaccharide SDLN1 is as follows:

2. The extraction method of the Bulbophyllum polysaccharide SDLN1 according to claim 1, characterized in that, The method comprises the following steps: 1) extracting defatted Bulbophyllum The dried Bulbophyllum sample is crushed, and petroleum ether is added for defatting treatment to obtain defatted Bulbophyllum; 2) extracting Bulbophyllum polysaccharide crude product The defatted Bulbophyllum extracted in step 1) is added to water, and then heated to reflux extraction; after the extraction is completed, the extract is concentrated to obtain a concentrated solution, and ethanol is added to the concentrated solution for alcohol precipitation to obtain a Bulbophyllum polysaccharide crude product; 3) extracting refined Bulbophyllum polysaccharide crude product The Bulbophyllum polysaccharide crude product extracted in step 2) is redissolved in water, and then subjected to Sevag method deproteinization, decolorization and dialysis treatment to obtain a refined Bulbophyllum polysaccharide crude product; 4) purification of the refined Bulbophyllum polysaccharide crude product The refined Bulbophyllum polysaccharide crude product extracted in step 3) is subjected to separation and purification treatment of DEAE-52 cellulose column and Sephadex G-100 column in sequence, and then freeze-dried to obtain the Bulbophyllum polysaccharide SDLN1.

3. The method of claim 2, wherein the method of extracting the Neottia nidus polysaccharide SDLN1 is characterized by, The method comprises the following steps: 1) extracting defatted Bulbophyllum 2Kg of dried Bulbophyllum is crushed into powder, petroleum ether is added at a material-liquid ratio of 1:20, and reflux extraction is performed at 90°C for 1h, and then filtration is performed to obtain defatted Bulbophyllum; 2) extracting Bulbophyllum polysaccharide crude product The filter residue of the defatted Bulbophyllum extracted in step 1) is dried, distilled water is added at a material-liquid ratio of 1:20, hot water reflux extraction is performed for 3 times, each time for 1h, and then filtration is performed; the filtrate is concentrated under reduced pressure to 2000mL, ethanol is added to make the alcohol content reach 80%, and then static setting is performed overnight, and the precipitate is collected by centrifugation and vacuum freeze-dried to obtain a Bulbophyllum polysaccharide crude product; 3) extracting refined Bulbophyllum polysaccharide crude product The Bulbophyllum polysaccharide crude product extracted in step 2) is dissolved in distilled water, and then subjected to Sevag method deproteinization with chloroform:n-butanol=4:1, the polysaccharide solution is mixed with Sevag reagent at a volume ratio of 3:1, and then H2O2 method decolorization is performed, dialysis is performed to remove small molecular substances, the liquid medicine is collected, and vacuum freeze-drying is performed to obtain a refined Bulbophyllum polysaccharide crude product; 4) purification of the refined Bulbophyllum polysaccharide crude product 100mg of the refined Bulbophyllum polysaccharide crude product extracted in step 3) is dissolved in 10ml of water, and then added to a DEAE-52 cellulose column which has been equilibrated, and then eluted with NaCl gradient solution 0-0.5Mol / L in sequence, and then different gradient Bulbophyllum polysaccharide eluate is collected, concentrated, dialyzed and freeze-dried to obtain polysaccharide elution components; the polysaccharide components are tracked by phenol sulfuric acid method, and then the peak containing the polysaccharide is collected and freeze-dried, and according to the results of each elution component, the 0.1 NaCl elution part is selected for further purification by gel purification system, the polysaccharide components are collected, concentrated, dialyzed and freeze-dried to obtain the Bulbophyllum polysaccharide SDLN1.

4. The application of the Bulbophyllum polysaccharide SDLN1 in claim 1 in the preparation of an anti-inflammatory drug.

5. The application of the Bulbophyllum polysaccharide SDLN1 extracted by the extraction method in any one of claims 2-3 in the preparation of an anti-inflammatory drug.

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