Application of potentilla anserine polysaccharide in preparation of medicine for relieving immunosuppression caused by cyclophosphamide

Abstract

The invention discloses application of potentilla anserine polysaccharide in preparation of a medicine for relieving immunosuppression caused by cyclophosphamide, and belongs to the medical nutrition research category of health management and promotion in the medical health field in the technical field of biological medicine. The invention provides an application of potentilla anserine polysaccharide in preparation of a medicine for relieving immunosuppression caused by cyclophosphamide, and verifications prove that the potentilla anserine polysaccharide can relieve weight loss of mice caused by cyclophosphamide, improve the levels of IFN-gamma, IgA, IgM and TNF-alpha, reduce the level of IL-17, relieve spleen injury of mice caused by cyclophosphamide, improve the immunity of the mice caused by cyclophosphamide, improve the immunity of the mice caused by cyclophosphamide, improve the immunity of the mice caused by cyclophosphamide, improve the immunity of the mice caused by cyclophosphamide, and improve the immunity of the mice caused by cyclophosphamide. The integrity of the intestinal tract is kept. The novel immune protective agent based on the potentilla anserine polysaccharide is developed by combining the immunoregulation advantage of natural polysaccharide and the traditional Tibetan medicine value of the potentilla anserine, and has important cultural inheritance significance and scientific application prospects.

Description

Technical Field

[0001] The present invention relates to biopharmaceutical technology, basic medicine, food science, nutrition, food analysis and inspection, etc., and particularly relates to the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating immunosuppression caused by cyclophosphamide. Background Art

[0002] Cyclophosphamide (CTX), as a broad-spectrum anti-tumor alkylating agent, is widely used in chemotherapy regimens for various malignant tumors. It inhibits tumor cell proliferation by interfering with DNA cross-linking, but at the same time also has significant toxicity to rapidly dividing normal immune cells (such as T lymphocytes, B lymphocytes and neutrophils), resulting in bone marrow suppression, immune organ atrophy and immune factor secretion disorders, and ultimately leading to immune system dysfunction in patients. According to statistics, more than 60% of patients receiving CTX chemotherapy will experience varying degrees of immunosuppression, manifested as an increased risk of opportunistic infection, weakened vaccine response and impaired anti-tumor immune response, which seriously restricts the clinical efficacy and affects the quality of life. Although immunomodulators such as recombinant human granulocyte colony-stimulating factor (G-CSF) and thymosin are often used clinically for intervention, these synthetic drugs have problems such as short half-life, the need for frequent injection administration, high cost, etc., and long-term use may induce autoimmune reactions or drug resistance, and there is an urgent need to develop a safer, longer-acting and more economical alternative solution.

[0003] In recent years, polysaccharide compounds of natural origin have attracted much attention due to their unique immunomodulatory activity and low toxicity. For example, lentinan, ganoderma polysaccharide, etc. have been proven to improve the immunosuppressive state through mechanisms such as activating the Toll-like receptor (TLR) signaling pathway, promoting macrophage phagocytic activity and enhancing NK cell killing ability. However, existing studies mostly focus on common medicinal fungi, and there are still significant gaps in the exploration of polysaccharide components of traditional Tibetan medicinal plants. Potentilla anserina (L.) Rydb., commonly known as "ginseng fruit", is recorded in Tibetan medicine and traditional Chinese medicine classics as having the effects of "tonifying qi and blood, benefiting the spleen and lungs". Modern pharmacological studies have shown that it is rich in polysaccharides, flavonoids and triterpenoid active ingredients, and among them, Potentilla anserina polysaccharide (PAP) shows the potential to stimulate lymphocyte proliferation and increase the secretion of IL-2 and IFN-γ in preliminary in vitro tests. However, there is currently no study systematically evaluating the repair effect of PAP on CTX-induced immunosuppression, and its action targets, in vivo pharmacodynamic characteristics and synergistic mechanism with chemotherapy drugs are not clear, which seriously limits its application and transformation in clinical immune adjuvant therapy. Summary of the Invention

[0004] The purpose of the present invention is to provide the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating immunosuppression caused by cyclophosphamide, so as to solve the problems existing in the above-mentioned prior art.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] One of the technical solutions of the present invention is the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating immunosuppression caused by cyclophosphamide.

[0007] Another technical solution of the present invention is the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating weight loss caused by cyclophosphamide.

[0008] A third technical solution of the present invention is the application of Potentilla anserina polysaccharide in the preparation of a drug for increasing the levels of IFN-γ, IgA, IgM, and TNF-α.

[0009] A fourth technical solution of the present invention is the application of Potentilla anserina polysaccharide in the preparation of a drug for reducing the level of IL-17.

[0010] A fifth technical solution of the present invention is the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating spleen injury caused by cyclophosphamide.

[0011] A sixth technical solution of the present invention is the application of Potentilla anserina polysaccharide in the preparation of a drug for maintaining intestinal integrity.

[0012] Based on the above technical solutions, the present invention has the following technical effects:

[0013] The present invention provides the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating immunosuppression caused by cyclophosphamide, and through experiments, it is confirmed that Potentilla anserina polysaccharide can alleviate weight loss in mice caused by cyclophosphamide, increase the levels of IFN-γ, IgA, IgM, and TNF-α, reduce the level of IL-17, alleviate spleen injury in mice caused by cyclophosphamide, and maintain intestinal integrity. Combining the immunomodulatory advantages of natural polysaccharides with the traditional medicinal value of Potentilla anserina, the development of a new type of immunoprotectant based on Potentilla anserina polysaccharide has important scientific significance and application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the following described drawings are only some embodiments of the present invention, and for those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0015] Figure 1 It is a graph of the weight change of mice and a technical roadmap. Among them, (A) is the flow chart of the animal experiment; (B) is the graph of the weight change of mice; (C) is the representative image of the spleen.

[0016] Figure 2It is a graph of the results of immune organ indices. Among them, (A) spleen index; (B) thymus index. *p < 0.05, **p < 0.01, ***p < 0.001 compared with the blank group; #p < 0.05, ##p < 0.01, p < 0.001 compared with the model group.

[0017] Figure 3 It is the effect of Potentilla anserina polysaccharide on serum inflammatory factors and immunoglobulins in mice. Among them, (A) IFN-γ; (B) IgA; (C) IgM; (D) IL-17; (E) IL-22; (F) TNF-α, *p < 0.05, **p < 0.01, ***p < 0.001 compared with the blank group; #p < 0.05, ##p < 0.01, p < 0.001 compared with the model group.

[0018] Figure 4 It is a graph of the HE staining results of mouse spleen and ileum. Among them, (A) representative graph of HE staining of mouse spleen; (B) quantification of white pulp area of spleen; (C) representative graph of HE staining of mouse ileum; (D) ratio of ileum villus height / crypt depth. *p < 0.05, **p < 0.01, ***p < 0.001 compared with the blank group; #p < 0.05, ##p < 0.01, p < 0.001 compared with the model group.

[0019] Figure 5 It is the effect of Potentilla anserina polysaccharide on short-chain fatty acids in mouse cecal contents. Among them, *p < 0.05, **p < 0.01, ***p < 0.001 compared with the blank group; #p < 0.05, ##p < 0.01, p < 0.001 compared with the model group. Detailed implementation manners

[0020] Now, various exemplary implementation manners of the present invention will be described in detail. This detailed description should not be regarded as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and implementation schemes of the present invention.

[0021] It should be understood that the terms described in the present invention are only for describing specific implementation manners and are not used to limit the present invention. In addition, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each intermediate value within any stated value or stated range and each smaller range between any other stated value or intermediate value within the stated range are also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0022] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although this invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the implementation or testing of this invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to the said documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0023] Without departing from the scope or spirit of this invention, various improvements and changes can be made to the specific embodiments of the specification of this invention, which are obvious to those skilled in the art. Other embodiments obtained from the specification of this invention are obvious to those skilled in the art. The specification and examples of this application are merely exemplary.

[0024] Regarding the use of "comprising", "including", "having", "containing", etc. herein, they are all open-ended terms, meaning including but not limited to.

[0025] The technical solutions described in this invention, unless otherwise specified, are all conventional solutions in the art. The reagents or raw materials used, unless otherwise specified, are all purchased from commercial channels or have been made public.

[0026] The embodiments of this invention provide the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating immunosuppression caused by cyclophosphamide.

[0027] In some specific embodiments, the preparation method of the Potentilla anserina polysaccharide comprises the following steps:

[0028] (1) After Potentilla anserina is crushed and defatted, it is enzymatically hydrolyzed with amyloglucosidase, and crude polysaccharide is obtained by alcohol precipitation.

[0029] (2) The crude polysaccharide is purified by column chromatography to obtain the Potentilla anserina polysaccharide.

[0030] In some specific embodiments, the method of enzymatically hydrolyzing with amyloglucosidase is as follows:

[0031] ① Water bath heating at 80 - 100 °C for 2 - 3 h;

[0032] ② 0.05% amyloglucosidase enzyme solution, at 50 - 65 °C for 1.5 h;

[0033] ③ Inactivating the enzyme at 100 °C.

[0034] The embodiments of this invention also provide the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating weight loss caused by cyclophosphamide.

[0035] The embodiments of the present invention also provide the application of Potentilla anserina polysaccharide in the preparation of a drug for increasing the levels of IFN-γ, IgA, IgM, and TNF-α.

[0036] The embodiments of the present invention also provide the application of Potentilla anserina polysaccharide in the preparation of a drug for reducing the level of IL-17.

[0037] The embodiments of the present invention also provide the application of Potentilla anserina polysaccharide in the preparation of a drug for alleviating the spleen injury caused by cyclophosphamide.

[0038] The embodiments of the present invention also provide the application of Potentilla anserina polysaccharide in the preparation of a drug for maintaining the integrity of the intestine.

[0039] Example 1

[0040] 1.1 Main reagents

[0041] Potentilla anserina: Tibet Autonomous Region; Cyclophosphamide: Sigma Company, USA; Lentinan: Hubei Guangren Pharmaceutical Co., Ltd.; ELISA kit: Jiangsu Enzyme Immunoassay Industry Co., Ltd.

[0042] 1.2 Main instruments

[0043] Low-temperature high-speed centrifuge, Eppendorf Company, Germany; Microplate reader, Thermo Fisher Scientific; Gas chromatograph, Shimadzu Corporation, Japan; -80°C ultra-low temperature refrigerator, Haier Biomedical Co., Ltd.; Electronic analytical balance, Sartorius Technology Co., Ltd., Germany.

[0044] Preparation method of Potentilla anserina polysaccharide:

[0045] (1) Clean Potentilla anserina, place it in a constant temperature drying oven at 40°C for drying, crush it and pass through a 60-mesh sieve;

[0046] (2) Take the Potentilla anserina powder crushed to 60 meshes, wrap it with filter paper, use petroleum ether as a degreasing agent, perform Soxhlet extraction at 180°C for 6 h, and dry it at low temperature after the petroleum ether has completely volatilized for standby.

[0047] (3) Add distilled water to the Potentilla anserina powder according to a solid-liquid ratio of 1:30, place it in a water bath at 85°C for 3 h to fully swell the degreased Potentilla anserina powder; after taking out the pot and cooling, adjust the pH to 4.5, add 0.05% amyloglucosidase enzyme solution, react at 55°C for 1.5 h; raise the temperature to 100°C to inactivate the enzyme for 20 min, then take out the pot and immediately place it in a cold water bath for cooling, centrifuge the extract at 5000 r / min for 10 min, perform suction filtration, and separate the supernatant and the solid residue.

[0048] (3) Take the above supernatant, add 5 times its volume of 95% ethanol for ethanol precipitation for 24 h; centrifuge at 4800 r / min for 10 min to separate the precipitate, remove the ethanol, add distilled water until the precipitate dissolves; rotary evaporate to remove the ethanol; dialyze with a dialysis bag for 48 h (replace the distilled water every 4 h); rotary evaporate and concentrate, pour into a plate, freeze-dry to obtain crude polysaccharide.

[0049] (4) Take the above solid residue, with a solid-liquid ratio of 1:1 of 95% ethanol, stir, let stand for 10 min, filter by suction, and take the precipitate; with a solid-liquid ratio of 1:1 of 70% ethanol, stir, let stand for 10 min, filter by suction, take the precipitate and add water, centrifuge at 4800 r / min for 10 min, then take the precipitate and add 95% ethanol, let stand for 10 min, filter by suction, and dry the precipitate to obtain pure Potentilla anserina polysaccharide.

[0050] 1.3 Experimental animals

[0051] Experimental animals: Female Balb / c mice, 5 weeks old, with a body weight of (20 ± 3) g, purchased from Zhuhai Besttone Biotechnology Co., Ltd., license number SCXK (Guangdong) 2020-0051; raised in the SPF-class mouse house of the Teaching Animal Hospital of Guangdong Ocean University. Clean the mouse cages every two days and change the bedding once. Feed and drink freely. The environmental temperature is 20 - 25 °C, the humidity is 40% - 70%, natural ventilation and air exchange, and a 12-h light-dark cycle.

[0052] 1.4 Experimental methods

[0053] 1.4.1 Grouping of mice and administration methods

[0054] Forty-eight female BALB / c mice were randomly divided into 6 groups after 7 days of adaptation, namely the control group, the model group, the positive group, the low-dose polysaccharide group, the medium-dose polysaccharide group, and the high-dose polysaccharide group, with 8 mice in each group. Different substances were intragastrically administered continuously for 21 days from the 8th day to the 28th day.

[0055] The control group and the model group were intragastrically administered normal saline;

[0056] The positive group was intragastrically administered lentinan at 100 mg / kg;

[0057] The polysaccharide groups were intragastrically administered different doses of Potentilla anserina polysaccharide (40 mg / kg, 80 mg / kg, 160 mg / kg).

[0058] Intragastric administration was performed once a day. During the experiment, the body weights of the mice were measured every day, and the states of the mice were observed.

[0059] On days 26, 27, and 28, the control group of mice was intraperitoneally injected with normal saline, and the other groups of mice were intraperitoneally injected with 80 mg / kg of cyclophosphamide to construct an immune deficiency model. On the evening of day 28, mouse feces were collected. On day 29, mouse blood was collected and the mice were sacrificed. After sacrificing the mice, the spleens, thymuses, cecal contents, and ileum tissues of the mice were collected, placed in cryotubes, and stored in a -80 °C refrigerator.

[0060] 1.4.2 Calculation of immune organ indices

[0061] Thymus index (mg / g) = thymus weight (mg) / live body weight of mouse (g);

[0062] Spleen index (mg / g) = spleen weight (mg) / live body weight of mouse (g).

[0063] 1.4.3 Serum cytokines and immunoglobulins

[0064] The contents of immunoglobulin A (IgA), immunoglobulin M (IgM), interleukin-17 (IL-17), interleukin-22 (IL-22), tumor necrosis factor (TNF-α), and interferon γ (IFN-γ) in the sample serum were measured according to the instructions of the ELLSA kit.

[0065] 1.4.4 HE staining of spleen and ileum

[0066] HE staining was entrusted to Wuhan Pinuofei Technology Co., Ltd. The experimental steps were roughly as follows: dewaxing paraffin sections to water → hematoxylin staining → eosin staining → dehydration and mounting.

[0067] 1.4.5 Determination of short-chain fatty acids

[0068] Weigh 50 mg of mouse feces, add 50 μL of pre-cooled 15% phosphoric acid after grinding, then add 1 ml of ethyl acetate, vortex for 1 min, then centrifuge (12000 rpm, 4 °C, 10 min), and take the supernatant through a membrane (pore size 0.22 μm) for gas chromatography analysis.

[0069] 2. Results and analysis

[0070] 2.1 Changes in mouse body weight

[0071] Cyclophosphamide (CTX) is a broad-spectrum anti-tumor drug used to treat various cancers, autoimmune, and immune-mediated diseases. However, taking CTX can also induce various adverse reactions, including DNA and immune cell damage, inhibition of cellular and humoral immune responses, disruption of the intestinal mucosal barrier, and intestinal flora translocation.

[0072] During the experiment, all mice were randomly divided into 6 groups, with 8 mice in each group. The grouping and gavage conditions were as follows Figure 1As shown in (A). The body weights of the mice were measured before daily intragastric administration, and the intragastric administration volume was adjusted according to the body weight. Figure 1 (B) is a graph showing the body weight changes of the mice during the 21-day intragastric administration. Starting from the 19th day, after continuous intraperitoneal injection of CTX for three days, the body weights of each group decreased. The body weights of the control group, model group, positive group, low-dose polysaccharide group, medium-dose polysaccharide group, and high-dose polysaccharide group decreased by 5.99%, 9.07%, 7.70%, 8.82%, 5.57%, and 4.71% respectively. It was found from the body weight changes that the degree of body weight reduction in the PAP group mice was alleviated, and the medium and high-dose groups in the PAP group had stronger alleviation effects on the body weight reduction of the mice than the positive group, proving that Potentilla anserina polysaccharide can improve the body of CTX-induced immunosuppressed mice.

[0073] 2.2 Physical images of the spleen

[0074] The spleen is an important immune organ. A smaller spleen is usually accompanied by a decline in immune function, including reduced antibody production and decreased immune cell activity. Figure 1 (C) is the physical image of the spleen of the mice after they were sacrificed 21 days after intragastric administration. Compared with the Con group, the spleen of the mice in the Mod group was significantly smaller, indicating that CTX caused severe damage to the mouse body and the immunosuppression model was successfully established. Figure 1 In (C), the spleen of the mice in the PAP group was significantly larger than that in the Mod group, indicating that the PAP group mice had an improvement effect on the body of CTX-induced immunosuppressed mice.

[0075] 2.3 Immune organ index

[0076] Immune organs can play a role in protecting the body from infection, and their immunomodulatory activities are closely related to changes in the immune organ index. The results of the immune organ index are as Figure 2 shown. Compared with the Con group, the immune organ index of the Mod group decreased significantly (p < 0.001), indicating that CTX caused atrophy of the spleen and thymus in the mice and the immunosuppression model was successfully established. Compared with the Mod group, there was no significant difference in the immune organ index of the PAP group (p > 0.05). It was speculated that the reason was that the PAP group did not have enough time to recover, resulting in the immune organs being affected and unable to reach the ideal state.

[0077] 2.4 Serum cytokines and immunoglobulins

[0078] Th1 and Th17 cells are two important cell subtypes differentiated from CD4+ T cells. Th1 cells can secrete TNF-α and IFN-γ cytokines, while Th17 cells mainly secrete IL-17 and IL-22. Cytokines are one of the most important effector and messenger molecules in the immune system. TNF-α and IFN-γ are crucial in protecting organisms from pathogens and activating the intestinal anti-infection response. IL-17 and IL-22 can induce innate immune responses in intestinal epithelial cells and play important roles in host defense and tissue repair on the intestinal mucosal surface. Immunoglobulins, as important markers of humoral immunity, play a fundamental role in immune regulation by directly neutralizing pathogens, transforming them into forms more easily consumed by phagocytes, and enhancing the ability of chemicals to destroy pathogens. Immunoglobulin levels are often used to evaluate humoral immune function. To evaluate whether CTX can also inhibit cytokine secretion and explore the regulatory role of polysaccharides, the secretion levels of TNF-α, IFN-γ, IL-17, IL-22, IgA, and IgM were measured.

[0079] The results of cytokines and immunoglobulins are as Figure 3 shown. Compared with the Con group, the levels of IFN-γ, IgA, IgM, IL-22, and TNF-α in the serum of mice in the Mod group were significantly decreased (p < 0.001), and the level of IL-17 in the serum was significantly increased (p < 0.001). Compared with the Mod group, the PAP group significantly increased the levels of IFN-γ, IgA, IgM, and TNF-α in the serum (p < 0.001), decreased the level of IL-17 in the serum, but the increase in IL-22 in the serum was not significant (p > 0.05). For the increase in the concentration of IL-17 in the model group, it is speculated that an immune compensatory response may have occurred. In the cyclophosphamide-induced model, the overall immune system is inhibited, but at the same time, in order to resist the invasion of external pathogens, the body locally enhances the immune response by increasing IL-17.

[0080] 2.5 HE staining of spleen and ileum

[0081] The spleen, as an important immune organ, is the site of the body's immune response, and the results of its HE staining are often used as a preliminary index for evaluating the immunopharmacology of drugs. The pathological staining results of the mouse spleen are as Figure 4 shown in A. The Con group had more white pulp areas, and the boundary between the white pulp and red pulp was clear. In the Mod group, there was no obvious boundary between the red pulp and white pulp of the spleen, and the volume of the white pulp was significantly reduced. In the LNT group and the PAP group, a significant recovery of the white pulp area was observed, indicating that the spleen damage caused by CTX was alleviated. The white pulp in the spleen is composed of dense lymphocytes and is the main site of specific immunity in the body. The quantitative results of the white pulp areas in the spleens of each group are asFigure 4 As shown in B of the figure, compared with the Con group, the white pulp area of the Mod group decreased significantly (p < 0.001), indicating that cyclophosphamide caused damage to the spleens of mice. Compared with the Mod group, the white pulp area of the PAP group increased significantly (p < 0.01), suggesting that PAP had the ability to alleviate the spleen damage caused by cyclophosphamide.

[0082] Maintaining intestinal integrity is a prerequisite for the normal function of the intestinal barrier. Intestinal villus length and crypt depth are often used as indicators of intestinal morphology. The pathological staining results of the ileum of mice are as Figure 4 shown in C. It can be found that the intestinal villi of mice in the Con group were intact and the crypt structure was clear. Compared with the Con group, the intestinal villi in the Mod group were ruptured, the crypts were shallower, and the structure was looser than that in the Con group. Compared with the Mod group, the intestinal villi in the PAP group were more intact and the crypt structure was clearer. The intestinal barrier damage caused by CTX was well alleviated, indicating that PAP had an ideal protective effect on the intestinal barrier. The ratio of ileal villus height / crypt depth in each group is as Figure 4 shown in D. Compared with the Con group, the ratio of ileal villus height / crypt depth in the Mod group decreased significantly (p < 0.001), indicating that CTX caused obvious damage to the intestinal barrier. Compared with the Mod group, the ratio of ileal villus height / crypt depth increased significantly after PAP intervention (p < 0.05), and the repair effect of the medium and high dose groups in the PAP group was more obvious (p < 0.001), with no significant difference from the Con group, indicating that PAP could improve the repair of the intestinal barrier.

[0083] 2.6 Determination of SCFAs

[0084] Natural polysaccharides that are difficult to digest by the human body are mainly degraded by intestinal microorganisms in the colon, thereby producing various metabolites (such as SCFAs), regulating the intestinal microbiota, and maintaining the balance of the intestinal microenvironment. SCFAs are mainly composed of acetic acid, propionic acid, butyric acid, valeric acid, isobutyric acid, and isovaleric acid. They play a key role in the establishment of the intestinal microbiota and the intestinal immune system and in maintaining the balance between them. The determination results of short-chain fatty acids in the cecal contents of mice are as Figure 5 shown. Compared with the Con group, the contents of acetic acid, butyric acid, isobutyric acid, valeric acid, and total SCFAs in the Mod group decreased significantly (p < 0.05). Compared with the Mod group, the concentrations of acetic acid and total SCFAs in the cecal contents were increased in the PAP group (p < 0.05). Among them, the concentrations of butyric acid, isobutyric acid, isovaleric acid, and valeric acid in the cecal contents of mice were significantly increased in the H-PAP group (p < 0.05). Therefore, these results suggest that PAP may be degraded by intestinal microorganisms in the colon to produce SCFAs, increasing the concentration of SCFAs in the cecum, thereby repairing the intestinal immune disorder and intestinal barrier damage induced by CTX.

[0085] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, rather than limitations on the implementation manners of the present invention. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to enumerate all implementation manners here. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the claims of the present invention.

Claims

1. Application of Potentilla anserina polysaccharide in the preparation of drugs for alleviating immunosuppression caused by cyclophosphamide.

2. The use according to claim 1, characterized in that: The preparation method of Potentilla anserina polysaccharide comprises the following steps: (1) After the bracken anserine is crushed and defatted, it is enzymatically hydrolyzed by amyloglucosidase and precipitated with alcohol to obtain crude polysaccharide; (2) The crude polysaccharide is purified by column chromatography to obtain the Potentilla anserina polysaccharide.

3. The use according to claim 2, characterized in that: The method for enzymolysis using amyloglucosidase is: ①Heat in a water bath at 80-100℃ for 2-3h; ②0.05% amyloglucosidase solution, 50-65°C, 1.5h; ③Inactivate the enzyme at 100℃.

4. Application of Potentilla anserina polysaccharide in the preparation of drugs for alleviating weight loss caused by cyclophosphamide.

5. Application of Potentilla anserina polysaccharide in the preparation of drugs for increasing the levels of IFN-γ, IgA, IgM, and TNF-α.

6. Application of Potentilla anserina polysaccharide in the preparation of drugs for reducing IL-17 levels.

7. Application of Potentilla anserina polysaccharide in the preparation of drugs for alleviating spleen damage caused by cyclophosphamide.

8. Application of Potentilla anserina polysaccharide in the preparation of medicines for maintaining intestinal integrity.

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