阅读说明：本技术 一种姜多糖及其制备方法与应用 (Ginger polysaccharide and preparation method and application thereof ) 是由 王一涛 王胜鹏 郝薇 于 2021-08-10 设计创作，主要内容包括：本发明公开了一种姜多糖及其制备方法与应用,属于中药提取物技术领域。该方法包括以下步骤：姜根茎经干燥、粉碎,乙醇回流脱脂,加水提取,加入乙醇醇沉,离心得到粗多糖。进而可采用超滤得到纯化姜多糖。该姜多糖的制备方法简单、易操作、成本低,可实现产业化,能维持溃疡性结肠炎小鼠的体重、降低疾病活动指数、增加结肠长度、降低结肠组织炎症因子水平,具有缓解结肠炎的作用,可制备成食品或治疗结肠炎的药物。(The invention discloses ginger polysaccharide and a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicine extracts. The method comprises the following steps: drying and crushing ginger rhizome, refluxing and degreasing with ethanol, adding water for extraction, adding ethanol for alcohol precipitation, and centrifuging to obtain crude polysaccharide. Further, ultrafiltration can be adopted to obtain purified ginger polysaccharide. The preparation method of the ginger polysaccharide is simple, easy to operate and low in cost, can realize industrialization, can maintain the body weight of an ulcerative colitis mouse, reduce disease activity index, increase colon length and reduce colon tissue inflammatory factor level, has the function of relieving colitis, and can be prepared into food or medicaments for treating colitis.)

1. The preparation method of the ginger polysaccharide is characterized by comprising the following steps: extracting rhizome of Zingiber officinale with water, performing solid-liquid separation for the first time, collecting liquid phase, precipitating with ethanol, and collecting precipitate.

2. The method of claim 1, wherein the water extraction is performed at 70-100 ℃ for 1-3 hours;

preferably, in the water extraction process, the material-liquid ratio of the ginger rhizome to water is 1: 5-40.

3. The method of claim 2, wherein prior to the water extraction, the method further comprises subjecting the ginger rhizome to a degreasing treatment;

preferably, the degreasing treatment is to perform reflux degreasing on the ginger rhizome and an ethanol water solution;

preferably, the concentration of ethanol in the ethanol aqueous solution is 70-100 vt%;

preferably, the ginger rhizome is dried and crushed and then degreased;

preferably, when the ginger rhizome is fresh ginger rhizome, the ginger rhizome is dried in an oven at 45-60 ℃ for 1-5 days and then crushed.

4. The process according to claim 1, wherein the alcohol used for the alcohol precipitation is ethanol;

preferably, the liquid phase is concentrated and then subjected to alcohol precipitation;

preferably, the volume of the concentrate after concentration is between 5 and 40 vt% of the liquid phase;

preferably, the alcohol is used in an amount of 3 to 6 times the volume of the concentrate during the alcohol precipitation.

5. The method as claimed in claim 4, wherein the second solid-liquid separation is performed by centrifugation at 4500g 2000-60 min;

preferably, before solid-liquid separation, the method also comprises standing the substance after alcohol precipitation for 30min-12 h.

6. The preparation method according to claim 5, further comprising purifying the precipitate obtained by the second solid-liquid separation to obtain purified gingerose;

preferably, the purification is carried out in an ultrafiltration mode, and the retention solution obtained by ultrafiltration is purified ginger polysaccharide;

preferably, the ultrafiltration is centrifugation at 500-;

preferably, the ultrafiltration tubes used for ultrafiltration have a molecular weight cut-off of >3 kDa.

7. A ginger polysaccharide, characterized by being prepared by the preparation method of any one of claims 1 to 6;

preferably, the weight average molecular weight of the gingerose is 0.1-5.0 × 10⁶Da；

Preferably, the polydispersity of the ginger polysaccharide is 0.8-4.0;

preferably, the particle size of the ginger polysaccharide is 30.0-60.0 nm.

8. The use of the ginger polysaccharide according to claim 7, wherein the ginger polysaccharide is used for the preparation of a food product or a medicament for the treatment of colitis;

preferably, the ginger polysaccharide is used for preparing a medicament for treating ulcerative colitis.

9. A food product, wherein the starting material for the food product comprises the ginger polysaccharide of claim 8.

10. A medicament for the treatment of colitis, wherein the starting material comprises the zingiber officinale polysaccharide of claim 7.

Technical Field

The invention relates to the technical field of traditional Chinese medicine extracts, and particularly relates to ginger polysaccharide and a preparation method and application thereof.

Background

Ulcerative Colitis (UC) is a chronic disease of the digestive tract with inflammation and ulceration in the colon and rectum. The main symptoms at the onset include abdominal pain, diarrhea with bloody stools, and weight loss. The etiology of ulcerative colitis is not well defined, and the pathogenesis associated with it is autoimmune disease, genetics, alteration of the intestinal flora, and environmental factors. Current conventional formulations for ulcerative colitis, include aminosalicylic acid, corticosteroids, immunomodulators and monoclonal antibodies, but these often have side effects and poor clinical efficacy.

The ginger is rhizome of Zingiber officinale Rosc. Fresh rhizome is fresh ginger, which is said to enter five zang organs in Shen nong Ben Cao Jing to remove pathogenic wind, cold and heat, typhoid, headache, nasal obstruction, cough and adverse rise of qi; stopping vomiting, removing phlegm and descending qi, relieving exterior syndrome, dispelling cold, stopping vomiting and removing toxicity; the dried rhizome is rhizoma Zingiberis, has effects of warming spleen and stomach for dispelling cold, restoring yang for dredging collaterals, warming lung for resolving fluid retention, and can be used for treating abdominal psychroalgia, emesis diarrhea, cold limbs, slight pulse, cold drink, cough and asthma. The ginger and the dried ginger are important traditional Chinese medicines commonly used by famous families in all generations, and the classic famous prescriptions, such as Sini decoction, dried ginger and monkshood decoction, coptis decoction, Baoyuan decoction and the like, contain the dried ginger or the ginger. Ginger contains a large amount of volatile oil, gingerol (gingerol, zingiberone, etc.), diphenyl heptane, carbohydrate, protein, vitamins and mineral elements. However, the research on ginger mainly focuses on the volatile oil and gingerol components, and the research on other components such as gingerol is only reported.

In view of this, the invention is particularly proposed.

Disclosure of Invention

One of the objects of the present invention is to provide a method for preparing ginger polysaccharide, which is simple, easy to operate, and low in cost, and can effectively extract ginger polysaccharide from ginger (ginger or dried ginger).

The invention also aims to provide the ginger polysaccharide prepared by the preparation method.

The invention also aims to provide an application of the ginger polysaccharide, such as the preparation of food or medicines.

The fourth purpose of the invention is to provide a food with the raw material containing the ginger polysaccharide.

The fifth purpose of the invention is to provide a medicine for treating colitis, which contains the ginger polysaccharide as the raw material.

The invention can be realized as follows:

in a first aspect, the present invention provides a method for preparing ginger polysaccharide, comprising the following steps: extracting rhizome of Zingiber officinale with water, performing solid-liquid separation for the first time, collecting liquid phase, precipitating with ethanol, and collecting precipitate.

In an alternative embodiment, the water extraction is carried out at 70-100 ℃ for 1-3 h.

In an optional embodiment, in the water extraction process, the material-liquid ratio of the ginger rhizome to the water is 1: 5-40.

In an alternative embodiment, the water extraction further comprises degreasing the ginger.

In an alternative embodiment, the degreasing treatment is reflux degreasing of ginger rhizome with an ethanol aqueous solution.

In alternative embodiments, the concentration of ethanol in the aqueous ethanol solution is from 70 to 100 vt%.

In an alternative embodiment, ginger rhizome is dried and crushed before being degreased.

In an alternative embodiment, when the raw material is fresh ginger rhizome, the ginger rhizome is dried in an oven at 45-60 deg.C for 1-5 days and then pulverized.

In an alternative embodiment, the liquid phase is concentrated and then subjected to alcohol precipitation.

In an alternative embodiment, the volume of the concentrate after concentration is 5-40 vt% of the liquid phase.

In an alternative embodiment, the alcohol is used in an amount of 3 to 6 times the volume of the concentrate during the alcohol precipitation.

In an alternative embodiment, the second solid-liquid separation is centrifugation at 4500g 2000-.

In an optional embodiment, before solid-liquid separation, the method further comprises standing the substance after alcohol precipitation for 30min-12 h.

In an optional embodiment, the method further comprises purifying the precipitate obtained by the second solid-liquid separation to obtain the purified gingerol polysaccharide.

In an alternative embodiment, the purification is performed by ultrafiltration, and the retentate obtained by ultrafiltration is purified ginger polysaccharide.

In an alternative embodiment, the ultrafiltration is centrifugation at 500-.

In an alternative embodiment, ultrafiltration tubes used for ultrafiltration have a molecular weight cut-off of >3 kDa.

In a second aspect, the present invention provides a ginger polysaccharide, prepared by the method of any one of the preceding embodiments.

In an alternative embodiment, the weight average molecular weight of the gingerose is 0.1-5.0 × 10⁶Da。

In an alternative embodiment, the ginger polysaccharide has a polydispersity index of 0.8 to 4.0.

In an alternative embodiment, the ginger polysaccharide has a particle size of 30.0-60.0 nm.

In a third aspect, the present invention provides the use of a ginger polysaccharide according to the previous embodiments, for the preparation of a food product or a medicament for the treatment of colitis.

In an alternative embodiment, the gingerose is used in the preparation of a medicament for the treatment of ulcerative colitis.

In a fourth aspect, the present invention provides a food product whose starting material comprises the ginger polysaccharide of the preceding embodiments.

In a fifth aspect, the present invention provides a medicament for the treatment of colitis, the starting material of which comprises the ginger polysaccharide of the previous embodiments.

The beneficial effect of this application includes:

the preparation method of the ginger polysaccharide provided by the application is simple, easy to operate and low in cost, and the ginger polysaccharide can be effectively extracted from ginger roots and stems. The prepared ginger polysaccharide has the function of relieving ulcerative colitis at least, can maintain the body weight of a mouse with the ulcerative colitis, reduce the disease activity index, increase the length of the colon and reduce the level of inflammatory factors of colon tissues, and provides scientific basis for the application of the ginger polysaccharide in preparing food or medicaments for treating the colitis.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a high performance gel chromatography coupled multi-angle laser scattering and differential refractometer (HPSEC-MALLS-RID) spectrum of ginger polysaccharide of example 4;

FIG. 2 is a schematic diagram showing the molding and administration timing of the mice in example 5;

FIG. 3 is a graph of the effect of various concentrations of gingerose on the body weight of UC mice in example 5, wherein p represents<0.0001，^##Represents p<0.01；

FIG. 4 is a graph showing the effect of various concentrations of gingerol on UC mouse disease index value (DAI) in example 5,^#p represents<0.05，^####Represents p<0.0001；

FIG. 5 is a graph of the effect of various concentrations of gingerose on colon length in mice from example 5, where p is expressed as<0.0001，^###Represents p<0.001；

FIG. 6 shows the effect of different concentrations of gingerol on pathological changes of UC mouse colon tissue in example 5 (hematoxylin-eosin staining, x 200);

FIG. 7 shows the effect of different concentrations of gingerol on pathological changes of UC mouse colon tissue in example 5 (scarlet red staining, x 200);

FIG. 8 is a graph showing the effect of various concentrations of gingerose on TNF- α content in UC mouse colon tissue in example 5<0.05，^##Represents p<0.01；

FIG. 9 is a graph showing the effect of various concentrations of gingerose on the IL-1. beta. content of UC mouse colon tissue in example 5<0.01，^##Represents p<0.01；

FIG. 10 is a graph showing the effect of various concentrations of gingerose on the IL-6 content in UC mouse colon tissue in example 5<0.05，^#Represents p<0.05。

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The ginger polysaccharide provided by the present application, and the preparation method and application thereof are specifically described below.

The application provides a preparation method of ginger polysaccharide, which comprises the following steps: extracting rhizome of Zingiber officinale with water, performing solid-liquid separation for the first time, collecting liquid phase, precipitating with ethanol, and collecting precipitate. The "ginger" herein may be ginger (fresh ginger) or dried ginger.

Wherein the water extraction can be carried out at 70-100 deg.C for 1-3 h. Specifically, the temperature for the water extraction may be 70 ℃, 75 ℃, 85 ℃, 90 ℃ or 100 ℃, preferably 100 ℃ (that is, it is understood that the extraction with boiling water) or any other value within the range of 70-100 ℃. The time for water extraction can be 1h, 1.5h, 2h, 2.5h or 3h, and the like, and can also be any other value within the range of 1-3 h.

In the water extraction process, the ratio of rhizoma Zingiberis recens to water may be 1:5-40, such as 1:5, 1:10, 1:20, 1:25, 1:35 or 1:40, or any other value within 1: 5-40.

In a preferred embodiment, the water extraction is carried out at 100 ℃ for 2h, and the feed-liquid ratio of ginger rhizome to water is 1: 20.

In an alternative embodiment, before the water extraction, the ginger rhizome is subjected to a degreasing treatment. The defatting treatment may be reflux defatting of ginger rhizome with ethanol water solution. Wherein, the concentration of the ethanol in the ethanol water solution can be 70-100 vt%, such as 75 vt%, 80 vt%, 85 vt%, 90 vt%, 95 vt% or 100 vt%, etc., and can also be any other value within the range of 70-100 vt%. Preferably, the concentration of ethanol in the ethanol aqueous solution can be 80 vt%, and the fat-soluble small molecular substances in the ginger rhizome raw material can be sufficiently extracted within the preferable concentration range.

Preferably, ginger roots are dried and crushed and then degreased, and crushing can increase the contact area between the raw material and the extracting agent, so that the dissolution rate of the functional components is increased.

In an alternative embodiment, the alcohol used for alcohol precipitation is preferably ethanol. In specific operation, the liquid phase can be concentrated and then subjected to alcohol precipitation. Concentration may be by concentrating the liquid phase to about 20% by volume (e.g. 5-40 vt%) to give a concentrate. In reference, the alcohol may be used in an amount of 3-6 times, such as 3.5 times, 4 times, 4.5 times, 5 times, 5.5 times or 6 times, preferably 4 times, the volume of the concentrate during the alcohol precipitation process.

Subsequently, the alcohol precipitated material is allowed to stand for 30min-12h, such as 30min, 1h, 100min, 2h, 5h, 8h, 10h or 12h, preferably 12 h.

Further, after standing, the mixture was subjected to a second solid-liquid separation.

In an alternative embodiment, the second solid-liquid separation may be performed by centrifugation at 4500g (e.g. 2000g, 3000g or 4500g, preferably 4000g) for 5-60min (e.g. 5min, 10min, 20min, 30min, 40min, 50min or 60min, preferably 30 min).

The precipitate obtained after the second solid-liquid separation is ginger crude polysaccharide.

Preferably, the precipitate can be re-dissolved in water, and freeze-dried to obtain ginger crude polysaccharide easy to store.

Further, purifying the ginger crude polysaccharide, namely purifying the precipitate obtained by the second solid-liquid separation, so as to obtain the purified ginger polysaccharide.

In an alternative embodiment, the purification may be performed by ultrafiltration, and the retentate obtained by ultrafiltration is purified ginger polysaccharide.

For reference, the ultrafiltration can be performed by centrifugation at 500-3500g (e.g., 500g, 2500g, 3000g, 3500g, etc., preferably 2500g) for 5-90min (e.g., 5min, 10min, 20min, 50min, 70min, 90min, etc., preferably 30 min).

The molecular weight cut-off of the ultrafiltration centrifugal tube used for ultrafiltration is more than 3 kDa.

Specifically, the purification process can be referred to as follows: adding deionized water into the ginger crude polysaccharide to prepare a solution with the concentration of 1-20mg/mL, transferring the solution into an ultrafiltration centrifugal tube (the molecular weight cut-off is more than 3kDa, Millipore, Billerica, MA, USA), centrifuging for 5-90min by 500-3500g, discarding the filtrate, supplementing water for many times, centrifuging, tracking and detecting the sugar content in the filtrate by adopting a phenol-sulfuric acid method until the filtrate is basically detected to be sugar-free, and freeze-drying the cut-off solution to obtain the purified ginger polysaccharide (represented by GP).

It should be noted that the above purification steps in the present application do not exclude other conventional purification methods, and are not described herein in any greater detail.

Correspondingly, the application also provides the ginger polysaccharide prepared by the preparation method.

The weight average molecular weight of the obtained ginger polysaccharide is about 0.1-5.0 × 10 by analyzing with high performance gel chromatography combined with multi-angle laser scattering and differential refractometer⁶Da, the polydispersion coefficient of the gingerose is 0.8-4.0, and the particle diameter of the gingerose is 30.0-60.0 nm. In certain embodiments, the weight average molecular weight of the resulting ginger polysaccharide is about 1.217 × 10⁶Da, polydispersity of about 1.81, and particle size of about 44.6 nm.

In addition, the application also provides the application of the ginger polysaccharide, for example, the ginger polysaccharide is used for preparing food (such as functional food and the like) or medicaments for treating colitis.

In a preferred embodiment, the gingerose is used in the manufacture of a medicament for the treatment of ulcerative colitis.

Further, the application also provides a food, and the raw material of the food comprises the ginger polysaccharide. The above-mentioned food products may, by reference, include milk and dairy products, fat products, frozen drinks, food products, bakery products, meat products, egg products, nutritional products, functional foods or beverages and the like, and in addition, other types of food products are not excluded.

The application also provides a medicament for treating colitis (especially ulcerative colitis), which comprises the ginger polysaccharide. The ginger polysaccharide is used as a raw material of the medicine, so that the medicine has the advantages of safety, definite action and less adverse reaction.

The rhizoma Zingiberis recens polysaccharide has good antiinflammatory effect and immunoregulatory activity, can relieve and treat ulcerative colitis, maintain body weight of mouse with ulcerative colitis, reduce disease activity index, increase colon length, and reduce tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), and interleukin-6 (IL-6) content of colon tissue. The effects of the aforementioned gingerose on ulcerative colitis may be associated with modulation of inflammatory cytokines, intestinal flora and the immune system and adhesion to the surface of colonic ulcers. In addition, the ginger polysaccharide as a drug carrier has good controlled release effect, high affinity and biodegradability.

The gingerose provided by the application can relieve ulcerative colitis by regulating the level of immune cells, interfering a signal transduction pathway, regulating the secretion of inflammatory cytokines and the abundance of intestinal flora and protecting the immune barrier of intestinal mucosa, and can be combined with other medicines for comprehensive treatment.

The features and properties of the present invention are described in further detail below with reference to examples.

Example 1

The present example provides a method for preparing a dried ginger polysaccharide:

(1) drying and pulverizing rhizoma Zingiberis, defatting with 80 vt% ethanol water solution under reflux, adding boiling water to make the ratio of material to liquid 1:20, extracting for 2 hr, and concentrating under reduced pressure to 20% volume.

(2) Adding 4 times volume of ethanol into the concentrate after vacuum concentration for precipitation, standing for 12h, centrifuging for 30min at 4000g, retaining the precipitate, adding water for redissolution, and freeze-drying to obtain crude polysaccharide.

(3) Adding deionized water into the crude polysaccharide to prepare a solution with the concentration of 10mg/mL, transferring the solution into an ultrafiltration centrifugal tube (the molecular weight cut-off is more than 3kDa, Millipore, Billerica, MA, USA), centrifuging for 30min at 2500g, discarding filtrate, supplementing water for many times, centrifuging, tracking and detecting the sugar content in the filtrate by adopting a phenol-sulfuric acid method until the filtrate is basically free of sugar, and freeze-drying the cut-off to obtain purified dried Ginger Polysaccharide (GP).

Example 2

The present example provides a method for preparing a dried ginger polysaccharide:

(1) drying and pulverizing rhizoma Zingiberis, defatting with 70 v/t% ethanol water solution under reflux, adding water to make the ratio of material to liquid 1:5, extracting at 70 deg.C for 3 hr, and concentrating under reduced pressure to 5% volume.

(2) Adding 3 times volume of ethanol into the concentrate after vacuum concentration for precipitation, standing for 30min, centrifuging for 5min at 4500g, retaining the precipitate, adding water for redissolution, and freeze drying to obtain crude polysaccharide.

(3) Adding deionized water into the crude polysaccharide to prepare a solution with the concentration of 1mg/mL, transferring the solution into an ultrafiltration centrifugal tube (molecular weight cut-off: 3kDa, Millipore, Billerica, MA, USA), centrifuging for 5min at 3500g, discarding filtrate, supplementing water for several times, centrifuging, tracking and detecting the sugar content in the filtrate by adopting a phenol-sulfuric acid method until the filtrate is basically free of sugar, and freeze-drying the cut-off solution to obtain purified dried Ginger Polysaccharide (GP).

Example 3

The embodiment provides a preparation method of ginger polysaccharide, which comprises the following steps:

(1) drying and pulverizing rhizoma Zingiberis recens, defatting with 100 vt% ethanol water solution under reflux, extracting at 98 deg.C for 1 hr at a material-to-liquid ratio of 1:40, and concentrating under reduced pressure to 40% volume.

(2) Adding 6 times volume of ethanol into the concentrate after vacuum concentration for precipitation, standing for 6h, centrifuging for 60min at 2000g, retaining the precipitate, adding water for redissolution, and freeze-drying to obtain crude polysaccharide.

(3) Adding deionized water into the crude polysaccharide to prepare a solution with the concentration of 20mg/mL, transferring the solution into an ultrafiltration centrifugal tube (the molecular weight cut-off is more than 3kDa, Millipore, Billerica, MA, USA), centrifuging for 90min at 500g, discarding filtrate, supplementing water for several times, centrifuging, tracking and detecting the sugar content in the filtrate by adopting a phenol-sulfuric acid method until the filtrate is basically free of sugar, and freeze-drying the cut-off to obtain the purified Ginger Polysaccharide (GP).

Example 4

Molecular parameter and conformation analysis of polysaccharide of dried ginger

The molecular parameters and chain conformation in the solution of the dried ginger polysaccharide prepared in example 1 were measured by high performance gel exclusion chromatography-multi-angle laser light scattering method, and about 3mg of dried ginger polysaccharide was weighed and dissolved in 1.0mL of 0.9% (w/v) aqueous sodium chloride solution, and HPLC-MALLS-RID-DAD detection conditions were as follows: the mobile phase is 0.9% (w/v) of sodium chloride aqueous solution; the column temperature of the chromatographic column is 35 ℃; the flow rate is 0.5 mL/min; the chromatographic column Shodex Ohpak SB-806M HQ (300mm × 8.0mm, i.d.) gel column is connected in series with Shodex Ohpak SB-G6B (50mm × 6mm) gel column; the sample was taken in an amount of 100. mu.L and the UV absorption of the sample was detected in parallel with DAD.

The results are shown in FIG. 1: the weight average molecular weight Mw, polydispersity Mw/Mn and particle diameter of Zingiberis rhizoma polysaccharide are 1.217 × 10⁶Da, 1.81 and 44.6nm, has ultraviolet absorption, indicating glycoprotein.

Example 5

Therapeutic effect of gingerose on ulcerative colitis

(1) Animal(s) production

25 male SPF grade C57BL/6J mice, weighing 17-24 g, were provided by the Guangdong province medical laboratory animal center.

(2) Grouping and molding

Animals were randomly divided into 5 groups, control, model, positive control, low-dose of dried ginger polysaccharide, high-dose of dried ginger polysaccharide, 5 animals per group. The test was started after one week of acclimatization of all mice, and starting from day 0, the mice of the control group were drunk with distilled water, and the remaining groups were freely drunk with 3% Dextran Sulfate Sodium (DSS) solution for 10 days to induce ulcerative colitis model, and administered on days 1, 3, 5, 7, and 9, respectively (as shown in fig. 2). The dosages of the dried ginger polysaccharide are respectively 100mg/kg and 200mg/kg, the dosage of the positive control group is 100mg/kg mesalazine solution, and the blank control group and the model group are respectively filled with distilled water with corresponding volumes. The ginger polysaccharide was the ginger polysaccharide prepared in example 1.

(3) Mouse body weight and Disease Activity Index (DAI) evaluation

During the experiment, the weight, the fecal characters and the hematochezia condition of each group of mice are observed and recorded every day, and the DAI scoring of the mice is carried out, wherein the scoring standard is as follows: percent body mass loss, stool quality, and stool bleeding.

As can be seen from the body weight results (as shown in fig. 3), the body weight of the mice began to decrease from day 6, the body weight of the model group was already significantly lower than that of the blank group (p <0.0001) at day 10, and the body weight of the mice in the high dose zingiber polysaccharide group was significantly improved (p <0.01) compared to that of the model group.

From the DAI results of fig. 4, the group administered with zingiber officinale polysaccharide was significantly reduced (p <0.05, p <0.0001) and dose-dependent compared to the model group.

(4) Colon length determination in mice

After the mice were sacrificed, the mice were dissected, the entire colon was taken out from the cecum to the anus, the colon was washed with precooled PBS solution, stretched and laid flat in a tensionless state, and the colon length of each group of mice was measured.

The results are shown in FIG. 5: the colon length was significantly shortened in the model group mice compared to the blank group (p < 0.0001); the colon length was significantly longer (p <0.001) in the zingiber polysaccharide high dose group mice compared to the model group.

(5) Pathological change of colon tissue in mouse

The colon tissues close to the rectum of each group of mice are taken and fixed in 4% paraformaldehyde for 48 hours, and pathological changes of the colon tissues are observed under a microscope after hematoxylin-eosin (HE) staining and sirius red staining respectively.

The HE staining results are shown in FIG. 6, the colon tissue glands of the control mice are arranged regularly, goblet cells are not damaged, and the tissue structure is complete; the colon tissue of the model group has serious intestinal epithelial injury, crypt atrophy or deletion, cellular edema and a large amount of inflammatory cell infiltration; the epithelial injury and inflammatory cell infiltration of colon tissues of mice in each administration group are reduced, and the phenomena of cryptitis and necrosis are improved, wherein the colon tissues of the mice in a high-dose group of rhizoma zingiberis polysaccharide only have mild epithelial injury and a small amount of inflammatory cell infiltration. Fig. 7 is a result of sirius red staining, and it can be seen by comparison that after sirius red staining is performed, a model group shows red, intestinal fibrosis of a control group is severe, deposition amounts of collagen in a mucosal tissue and a submucosal layer of the mucosal tissue are gradually increased, and staining degrees and staining areas are increased; compared with a model group, the fibrosis condition of each administration group is relieved, and the fibrosis condition of colon tissues of a rhizoma zingiberis polysaccharide high-dose group is improved most obviously.

(6) Determination of contents of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) and interleukin-6 (IL-6) in colon tissue

Taking colon tissue 3cm, cleaning with precooled PBS solution, cutting into pieces, placing in RPMI1640 culture medium, incubating in a carbon dioxide incubator for 24h, centrifuging, taking tissue culture solution, and determining the contents of TNF-alpha, IL-1 beta and IL-6 according to the operation instruction of the ELISA kit.

The results are shown in fig. 8 to 10: compared with the blank group, the content of TNF-alpha, IL-1 beta and IL-6 in the model group is obviously increased (p <0.05, p <0.01 and p <0.05), and compared with the model group, the content of TNF-alpha, IL-1 beta and IL-6 in colon tissues can be obviously reduced in the rhizoma zingiberis polysaccharide high-dose group (p <0.01, p <0.01 and p < 0.05).

In summary, the preparation method of the ginger polysaccharide provided by the application is simple, easy to operate and low in cost, and the ginger polysaccharide can be effectively extracted from ginger rhizomes. The prepared ginger polysaccharide has the function of relieving ulcerative colitis at least, can maintain the body weight of a mouse with the ulcerative colitis, reduce the disease activity index, increase the length of the colon and reduce the level of inflammatory factors of colon tissues, and provides scientific basis for the application of the ginger polysaccharide in preparing food or medicaments for treating the colitis.

The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.