Lotus acidic polysaccharide as well as preparation process and new application thereof

Abstract

The invention discloses a lotus acidic polysaccharide in the technical field of extraction of plant polysaccharides, and the lotus acidic polysaccharide is mainly composed of rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose with the molar percentage of 4-10%: 1-3%: 60-85%: 5-10%: 3-8%. The polysaccharide is obtained by adopting a water extraction and alcohol precipitation method and anion exchange column separation and purification. Researches show that the lotus acidic polysaccharide can be used for preventing and / or treating hepatic fibrosis.

Description

Technical Field

[0001] The invention relates to the technical field of plant polysaccharide extraction, and in particular to lotus acidic polysaccharide, a preparation process and a new use thereof. Background Art

[0002] Fibrosis is a chronic and progressive pathological process characterized by abnormal increase or excessive deposition of extracellular matrix in tissues. Within a certain range of tissue cell damage, moderate fibrosis can help repair trauma. However, if this repair reaction is excessive, it will cause fibrosis in some organs, leading to decreased organ function, gradually destroying the organ tissue structure and causing organ sclerosis. Studies have found that fibrosis is mainly related to oxidative stress. Among many signaling pathways, transforming growth factor-β1 (TGF-β1) / Smad, high mobility group 1 / Toll-like receptor 4, cyclic adenosine monophosphate / protein kinase A (cAMP / PKA) and MMP (matrix metalloproteinase) / TIMPs (tissue inhibitors of metalloproteinases) systems contribute to the progression of fibrosis. Therefore, targeting these signaling pathways is likely to have a certain effect on the treatment of fibrosis. In addition, existing evidence shows that natural plant polysaccharides can regulate each stage of the fibrotic response and reduce the level of fibrosis in multiple organs, including primary damage to organs and activation of effector cells. Therefore, natural polysaccharides can be used as potential chemical agents with anti-fibrotic activity.

[0003] Lotus (Nelumbinis Flos) is the flower bud of the plant Nelumbo of the Nymphaeaceae family, which is produced in Hunan, Hubei, Fujian, Jiangsu, Zhejiang and other places. In ancient medical books, it has the effect of dispersing blood stasis and stopping bleeding, and is mainly used to treat vomiting blood due to falls, hematuria, metrorrhagia, and blister sores. As recorded in the Shennong's Herbal Classic, "it can promote blood circulation and stop bleeding, remove dampness and eliminate wind, clear the heart and cool blood, and relieve heat and detoxify." In modern pharmacological research, lotus has anti-tumor, hypoglycemic, antioxidant, and anti-inflammatory effects. Studies have found that the above wide range of activities are derived from its diverse active chemical substances, such as flavonoids, alkaloids, polysaccharides, phenols, and glycosides. However, there is little research on the mechanism of action of small and large molecular active substances in traditional Chinese medicine, and it is difficult to analyze the structure of large molecular active polysaccharides. These problems seriously affect the research on the drugability of lotus plants, and polysaccharides have attracted much attention as good active substances due to their good water solubility, rich content in plants, and high safety. Therefore, extracting the polysaccharide of the main active substance of lotus and analyzing its structure are not reported in the prior art. Summary of the invention

[0004] Aiming at the deficiencies of the prior art, the present invention provides a simple and effective process and method for extracting and purifying plant polysaccharides, designs a method for separating acidic polysaccharide LFM22 from lotus, and uses the acidic polysaccharide LFM22 to alleviate liver fibrosis.

[0005] One of the purposes of the present invention is to provide a lotus acid polysaccharide, which is mainly composed of rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose, wherein the molar percentages of rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose are 4-10%: 1-3%: 60-85%: 5-10%: 3-8%, respectively, and the lotus acid polysaccharide has a weight average molecular weight of 10-40 kDa and a dispersion coefficient D of 1-1.6.

[0006] Furthermore, the molar ratio of rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose is 5.99:1.43:78.65:6.74:3.9.

[0007] Furthermore, the sugar residues of the lotus acid polysaccharide are connected in the following ways: terminal-connected arabinose, terminal-connected rhamnose, 1,5-connected arabinose, 1,2-connected rhamnose, terminal-connected glucuronic acid, terminal-connected galactose, terminal-connected galacturonic acid, 1,3,5-connected arabinose, 1,2,4-connected rhamnose, 1,4-connected galactose, 1,3-connected galactose, 1,3,4-connected galacturonic acid, 1,4,6-connected galactose, 1,3,6-connected galactose and 1,4-connected galacturonic acid, and the molar ratios thereof are 1-3:1-2:1-2:1-3:1-2:1-3:1-3:1-2:1-3; 1-2:1-2:1-4:1-2:1-3:50-70.

[0008] Furthermore, the molar ratios of sugar residues of the lotus acidic polysaccharide are 2:1:1:2:1:2:2:1:2:1:1:3:1:2:60.

[0009] Furthermore, the lotus acid polysaccharide has the following structure:

[0010]

[0011] The second object of the present invention is to provide a method for preparing the lotus acid polysaccharide, comprising the following steps:

[0012] 1) Crushing: crush the lotus with a crusher to screen out lotus powder of 100-300 mesh;

[0013] 2) Swelling: immerse the lotus powder in water and swell at 70°C for 2 to 4 hours;

[0014] 3) Grinding: Grind with a colloid mill homogenizer for 2 to 4 times, and then use a high-pressure homogenizer to break the cell wall for 2 to 4 times;

[0015] 4) Decoction: Heat to keep slightly boiling and decoct for 2 to 4 hours / time, decoct twice in total, and combine the filtrate;

[0016] 5) Filtration: The filtrate is ultrafiltered using a membrane filter to obtain a fraction with a molecular weight greater than 5000 Da;

[0017] 6) Dialysis: The ultrafiltration fraction is concentrated to 1 / 10-1 / 20 of the original volume, cooled naturally to room temperature, and the concentrate is dialyzed against running water through cellophane for 2-3 days;

[0018] 7) Alcohol precipitation: The dialysate is concentrated to 1 / 5-1 / 10 of the original volume, cooled naturally to room temperature, centrifuged at 4000-8000 rpm for 10-30 min, and the supernatant is taken and 3-6 times the volume of the supernatant is added with 95% ethanol while stirring, and allowed to stand overnight;

[0019] 8) Freeze drying: centrifuge the alcohol precipitate at 4000-8000 rpm for 10-30 min, add water to the precipitate and heat to evaporate the remaining ethanol, freeze it and freeze dry it in a freeze dryer to obtain crude polysaccharide;

[0020] 9) Separation: The crude polysaccharide was separated by DEAE anion exchange and eluted with 0.2 M NaCl to obtain lotus acid polysaccharide.

[0021] Furthermore, the lotus material is selected as dried or fresh flower buds.

[0022] Furthermore, in step 2), the liquid-to-water ratio of water to lotus is 1:10 to 1:30;

[0023] Furthermore, the crude polysaccharide in step 9) is separated by anion exchange column DEAE FAST FLOW with a sample load ranging from 30 to 50 g and a concentration of 40 to 80 mg / mL.

[0024] The third object of the present invention is to provide the use of lotus acid polysaccharide in the preparation of drugs for treating liver fibrosis or drugs for assisting the treatment of liver fibrosis.

[0025] Specifically, the dosage concentration is 0.5 mg / mL to 1 mg / mL.

[0026] The fourth object of the present invention is to provide a pharmaceutical composition comprising the above-mentioned lotus acid polysaccharide and pharmaceutically acceptable excipients.

[0027] The invention separates and purifies a lotus acid polysaccharide LFM22 from lotus. The monosaccharide composition analysis by a PMP pre-column derivatization method shows that the lotus acid polysaccharide LFM22 contains rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose, and the molar percentages thereof are respectively in the range of 4-10%, 1-3%, 60-85%, 5-10% and 3-8%.

[0028] Experimental verification shows that lotus acid polysaccharide LFM22 has a significant inhibitory effect on the fibrosis of hepatic stellate cells LX-2, and the inhibition rate at 0.5 mg / mL to 1 mg / mL is between 50% and 80%. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is a schematic diagram of the HPGPC results of LFM22;

[0030] Figure 2 Schematic diagram of monosaccharide composition analysis of LFM22;

[0031] Figure 3 1H NMR (A) and 13C NMR (B) spectra of LFM22;

[0032] Figure 4 Schematic diagram of the initial screening effect of LFM22 (A160) on alleviating TGF-β-induced hepatic stellate cell LX-2 fibrosis;

[0033] Figure 5 Schematic diagram of the effect of different concentration gradients of (A160) LFM22 on alleviating TGF-β-induced hepatic stellate cell LX-2 fibrosis. DETAILED DESCRIPTION

[0034] The following is further described in detail through specific implementation methods:

[0035] 1. Polysaccharide extraction, separation and purification

[0036] 1) Crushing: crush and screen the dried / fresh lotus with a 100-300 mesh crusher;

[0037] 2) Swelling: immerse the lotus powder in water, with a solid-liquid ratio of 1:10-1:30 (g / mL), and swell at 70°C for 2-4 hours;

[0038] 3) Grinding: Grind with a colloid mill homogenizer for 2 to 4 times, and then use a high-pressure homogenizer to break the cell wall for 2 to 4 times;

[0039] 4) Decoction: Heat to keep slightly boiling and decoct for 2 to 4 hours / time, decoct twice in total, and combine the filtrate;

[0040] 5) Filtration: The filtrate is ultrafiltered using a membrane filter to obtain a fraction with a molecular weight greater than 5000 Da;

[0041] 6) Dialysis: The ultrafiltration fraction is concentrated to 1 / 10-1 / 20 of the original volume, cooled naturally to room temperature, and the concentrate is dialyzed against running water through cellophane for 2-3 days to remove pigments and small molecules. After the dialysis is completed, it is filtered through gauze;

[0042] 7) Alcohol precipitation: The dialysate is concentrated to 1 / 5-1 / 10 of the original volume, cooled naturally to room temperature, centrifuged at 4000-8000 rpm for 10-30 min, and the supernatant is taken and 3-6 times the volume of the supernatant is added with 95% ethanol while stirring, and allowed to stand overnight;

[0043] 8) Freeze drying: centrifuge the alcohol precipitate at 4000-8000 rpm for 10-30 min, add 0.5-3 L of water to reconstitute the precipitate, heat to evaporate the remaining ethanol, freeze it, and freeze dry it in a freeze dryer to obtain crude polysaccharide;

[0044] 9) Separation: The crude polysaccharide was separated by DEAE anion exchange, with the sample loading range of 30-50 g and the concentration of 40-80 mg / mL. The lotus acidic polysaccharide LFM22 was obtained by elution with 0.2 M NaCl.

[0045] 2. Analysis of polysaccharide purity, monosaccharide composition and sugar residue connection mode

[0046] 1. Determination of polysaccharide purity:

[0047] Accurately weigh 3-7 mg of polysaccharide sample, dissolve in 300-700 μL of 0.1 M sodium nitrate, shake to fully dissolve the sample, centrifuge at 4000-8000 rpm for 5-10 min, and filter the supernatant through a 0.22 μM aqueous filter membrane for purity determination.

[0048] Polysaccharide-specific gel chromatography columns in series: Waters UltrahydrogelTM 2000 and UltrahydrogelTM 500 in series. Chromatographic conditions: mobile phase 0.1M sodium nitrate, flow rate 0.5mL / min, injection volume 10-20μL, column temperature 35℃, UV absorption wavelength 280nm, differential detector temperature 35℃, 40-60min / sample.

[0049] 2. Analysis of polysaccharide monosaccharide composition:

[0050] 1) Complete acid hydrolysis: Weigh 2-4 mg of sample, dissolve in 2 mL of distilled water, vortex and oscillate to dissolve as much as possible (heating or ultrasound can be used), add the sample solution to a heart-shaped bottle (specification 50 / 19), then add 2 mL of 4M trifluoroacetic acid (TFA) and mix well, add a hollow stopper to the heart-shaped bottle, and seal the stopper and the contact port of the heart-shaped bottle with medical rubber paste, and heat and hydrolyze at 110°C for 2-5 hours. After hydrolysis, cool, add methanol and evaporate under reduced pressure for several times to remove TFA. Add 200 μL of distilled water to dissolve the hydrolyzate.

[0051] 2) PMP derivatization process: Take 100μL of the 200μL hydrolyzate from the previous step and add it to a 10mL EP tube, add 100μL of 0.6M NaOH solution and mix well. Add 200μL of freshly prepared 0.5M PMP, seal the tube and mix well, heat in a 70℃ water bath for 1h+40min, cool to room temperature after the reaction is completed, then add 200μL of 0.3M HCl, and then add 400μL of deionized water to make the total volume of the system 1mL.

[0052] 3) Preparation of standard solution: Prepare 9 mg / mL of each monosaccharide with deionized water, take 100 μL of each and mix them, then you will get 10 kinds of monosaccharide standard solutions, each with a concentration of 1 mg / mL. When derivatizing, take out 100 μL of the standard mixture for derivatization operation.

[0053] 4) Extraction: Oscillate for 3 minutes, centrifuge at 8000 rpm for 5 minutes, and stand at room temperature for 30 minutes. Keep the upper aqueous phase, repeat the extraction with chloroform for 3 times, and filter the upper aqueous phase through a 0.22 μm microporous membrane.

[0054] 5) HPLC analysis:

[0055] The analytical column was a C18 reverse phase column, the mobile phase was phosphate buffer (the volume ratio of pH 7.0 phosphate buffer to acetonitrile was 33:7), the column temperature was 35°C, the flow rate was 1 mL / min, the ultraviolet absorption wavelength was 245 or 254 nm, the injection volume was 10 μL, and the detection time was 1 h / sample.

[0056] 3. Analysis of the connection mode of sugar residues in polysaccharides

[0057] Take the lotus acid polysaccharide component LFM22 for methylation analysis. Weigh 5-12 mg of the sample in advance into a 50 mL chicken heart bottle, add 2 mL of deionized water to dissolve, freeze-dry and place in a drying cabinet overnight (the reaction is guaranteed to be anhydrous), and completely dissolve in 2 ml of anhydrous DMSO the next day. After the polysaccharide sample is completely dissolved (if the sample is not well soluble in DMSO, it can be heated in a 70 ° C oil bath and stirred overnight to dissolve), add 50 mg of ground dry sodium hydroxide powder, and stir the reaction at room temperature for 7 minutes. In an ice water bath, slowly add 1 mL of iodomethane (about 35 minutes), remove the ice bath, and react at room temperature for 30 minutes away from light. After the reaction is completed, add 1 ml of deionized water to the reaction system to quench the reaction. The solution is concentrated under reduced pressure to remove excess unreacted iodomethane, dialyze against deionized water for 24 to 72 hours, and freeze-dry the solution. Repeat 4 to 5 times again. The sample after the reaction was completely acid hydrolyzed, 4 ml of 2M TFA was added and reacted at 110°C for 4 h. After cooling to room temperature, methanol was added several times to remove excess acid until there was no sour taste. After the reaction, 2 ml of water and 50 mg of sodium borohydride were added for reduction. After sealing, the reaction was carried out at room temperature for 3 h. Sodium borohydride was neutralized with 25% acetic acid solution to terminate the reaction. Methanol was repeatedly added to remove excess acid, and the sample was placed in a 100°C oven for drying for 15 min. 3 ml of acetic anhydride was added and placed in a 100°C oven for acetylation. The reaction was carried out for 1.5 h. Toluene was repeatedly added to remove excess acetic anhydride. Finally, 15 mL of chloroform and 15 mL of deionized water (v / v was 1:1) were added to the reaction bottle, extracted, and the aqueous phase was discarded. The organic phase was washed with deionized water 5 times, dried over anhydrous sodium sulfate, and the chloroform was concentrated under reduced pressure to 200 μL, filtered through a 0.22 μm organic phase filter membrane, and placed in a liquid phase vial. The connection mode of polysaccharides was detected and analyzed by GC-MS (Thermo Fisher ISQ7000).

[0058] like Figure 1 , 2 As shown, Figure 1 , differential detection diagram of lotus acidic polysaccharide LFM22; Figure 2 , the peak order of monosaccharide composition measured by PMP pre-column derivatization of 10 monosaccharide standards (Man: mannose; Gul: gulose; Rha: rhamnose; GlcA: glucuronic acid; GalA: galacturonic acid; Glc: glucose; Gal: galactose; Xyl: xylose; Ara: arabinose; Fuc: fucose) and LFM22 monosaccharide composition analysis.

[0059] HPGPC purity analysis showed that the weight average molecular weight of lotus acid polysaccharide LFM22 was 10-40 kDa, and the dispersion coefficient D was 1-1.6.

[0060] The monosaccharide composition analysis by PMP pre-column derivatization method showed that the lotus acid polysaccharide LFM22 contained rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose, and the molar percentages ranged from 4-10%, 1-3%, 60-85%, 5-10% and 3-8%, respectively.

[0061] The analysis of the sugar residue connection patterns of lotus acid polysaccharide LFM22 by GC-MS showed that the following connection patterns were found: terminal-linked arabinose, terminal-linked rhamnose, 1,5-linked arabinose, 1,2-linked rhamnose, terminal-linked glucuronic acid, terminal-linked galactose, terminal-linked galacturonic acid, 1,3,5-linked arabinose, 1,2,4-linked rhamnose, 1,4-linked galactose, 1,3-linked galactose, 1,3,4-linked galacturonic acid, 1,4,6-linked galactose, 1,3,6-linked galactose and 1,4-linked galacturonic acid, and the molar ratios were 1-3:1-2:1-2:1-3:1-2:1-3:1-3:1-2:1-3; 1-2:1-2:1-4:1-2:1-3:50-70.

[0062] The details are shown in Table 1 below.

[0063] Table 1 is a schematic diagram of the sugar residue connection method of lotus acid polysaccharide LFM22

[0064]

[0065] 3. Determination of anti-liver fibrosis activity of lotus acid polysaccharide LFM22

[0066] 1. Initial screening and determination of the activity of lotus acidic polysaccharide LFM22:

[0067] In order to detect whether lotus acid polysaccharide has anti-liver fibrosis activity, we first verified whether the polysaccharide has the effect of inhibiting the activation of hepatic stellate cells LX-2 and reducing the deposition of extracellular matrix. Figure 4 The preliminary screening results showed that lotus acid polysaccharide LFM22 (A160) has a good anti-fibrosis effect.

[0068] 2. Western blot experiments were used to detect the effects of gradient concentrations of lotus acid polysaccharide LFM22 on the expression of α-SMA and Collagen I proteins in hepatic stellate cells LX-2:

[0069] Human hepatic stellate cells LX-2 were cultured in DMEM medium containing 10% fetal bovine serum (purchased from Gibco, USA), 100 U / mL penicillin and 100 μg / mL streptomycin in a volume ratio of 1:1. LX-2 cells in the logarithmic growth phase were taken and cultured at 1.2×10 6The cells were seeded at a density of 100 cells / well in a 6-well plate. After culturing for 24 hours, the cells were stimulated simultaneously with 0, 0.1, 0.5, and 1 mg / ml lotus acid polysaccharide LFM22 mixed with 0 and 10 ng / mL TGF-β1 protein. After culturing for 48 hours, the supernatant was discarded, the cells were rinsed with pre-cooled PBS, and the cells were lysed on ice for 10 minutes with an appropriate volume of lysis buffer (lysis buffer purchased from Thermo), and centrifuged at 4°C for 10 minutes. The supernatant was collected, 5× loading buffer was added, and the protein was denatured in a sample cooker for 10 minutes. After cooling, it was stored at -80°C. Immunoblotting was used to detect the protein expression of α-SMA and Collagen I. The results are as shown in Figure 5 As shown, lotus acidic polysaccharide LFM22 can significantly inhibit the expression of α-SMA and Collagen I proteins.

[0070] In summary, the test results of the embodiment show that the lotus acid polysaccharide LFM22 can significantly inhibit the expression of fibrosis-related proteins α-SMA and Collagen I without affecting the cell viability of hepatic stellate cells LX-2 in vitro, thereby inhibiting the activation of hepatic stellate cells. Therefore, the lotus acid polysaccharide LFM22 of the present invention can become a potential carbohydrate drug or liver protection health product for preventing and / or treating liver fibrosis.

[0071] The above is only an embodiment of the present invention, and the common knowledge such as the known specific structure and characteristics in the scheme is not described in detail here. It should be pointed out that for those skilled in the art, several deformations and improvements can be made without departing from the structure of the present invention, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the specification can be used to interpret the content of the claims.

Claims

1. A lotus acid polysaccharide, It is characterized in that The lotus acid polysaccharide is mainly composed of rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose, wherein the molar percentages of rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose are 4-10%: 1-3%: 60-85%: 5-10%: 3-8% respectively. The weight average molecular weight of the lotus acid polysaccharide is 10-40 kDa, and the dispersion coefficient D is 1-1.

6.

2. The lotus acid polysaccharide according to claim 1, Features: The molar ratio of rhamnose, glucuronic acid, galacturonic acid, galactose and arabinose is 5.99:1.43:78.65:6.74:3.

9.

3. The lotus acid polysaccharide according to claim 1, Features: The sugar residues of the lotus acid polysaccharide are connected in the following ways: terminal-connected arabinose, terminal-connected rhamnose, 1,5-connected arabinose, 1,2-connected rhamnose, terminal-connected glucuronic acid, terminal-connected galactose, terminal-connected galacturonic acid, 1,3,5-connected arabinose, 1,2,4-connected rhamnose, 1,4-connected galactose, 1,3-connected galactose, 1,3,4-connected galacturonic acid, 1,4,6-connected galactose, 1,3,6-connected galactose and 1,4-connected galacturonic acid, and the molar ratios thereof are 1-3:1-2:1-2:1-3:1-2:1-3:1-3:1-2:1-3; 1-2:1-2:1-4:1-2:1-3:50-70.

4. The lotus acid polysaccharide according to claim 3, Features: The molar ratios of sugar residues of the lotus acidic polysaccharide are 2:1:1:2:1:2:2:1:2:1:1:3:1:2:

60.

5. The lotus acid polysaccharide according to claim 1, Features: Lotus acid polysaccharide has the following structure:

6. The method for preparing lotus acid polysaccharide according to any one of claims 1 to 5, Features: The following steps are involved: 1) Crushing: crush the lotus with a crusher to screen out lotus powder of 100-300 mesh; 2) Swelling: immerse the lotus powder in water and swell at 70°C for 2 to 4 hours; 3) Grinding: Grind with a colloid mill homogenizer for 2 to 4 times, and then use a high-pressure homogenizer to break the cell wall for 2 to 4 times; 4) Decoction: Heat to keep slightly boiling and decoct for 2 to 4 hours / time, decoct twice in total, and combine the filtrate; 5) Filtration: The filtrate is ultrafiltered using a membrane filter to obtain a fraction with a molecular weight greater than 5000 Da; 6) Dialysis: The ultrafiltration fraction is concentrated to 1 / 10-1 / 20 of the original volume, cooled naturally to room temperature, and the concentrate is dialyzed against running water through cellophane for 2-3 days; 7) Alcohol precipitation: The dialysate is concentrated to 1 / 5-1 / 10 of the original volume, cooled naturally to room temperature, centrifuged at 4000-8000 rpm for 10-30 min, and the supernatant is taken and 3-6 times the volume of the supernatant is added with 95% ethanol while stirring, and allowed to stand overnight; 8) Freeze drying: centrifuge the alcohol precipitate at 4000-8000 rpm for 10-30 min, add water to the precipitate and heat to evaporate the remaining ethanol, freeze it and freeze dry it in a freeze dryer to obtain crude polysaccharide; 9) Separation: The crude polysaccharide was separated by DEAE anion exchange and eluted with 0.2 M NaCl to obtain lotus acid polysaccharide.

7. The preparation method according to claim 6, Features: In step 2), the liquid-to-water ratio of water to lotus is 1:10-1:30; in step 9), the crude polysaccharide is separated by anion exchange column DEAE FAST FLOW with a sample loading range of 30-50 g and a concentration of 40-80 mg / mL.

8. Use of the lotus acid polysaccharide according to any one of claims 1 to 5 in the preparation of a drug for treating liver fibrosis or a drug for assisting the treatment of liver fibrosis.

9. The use according to claim 8, Features: The dosage concentration is 0.5mg / mL to 1mg / mL.

10. A pharmaceutical composition comprising the lotus acid polysaccharide according to any one of claims 1 to 5, and pharmaceutically acceptable excipients.

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