阅读说明：本技术 石斛多糖及其制备方法 (Dendrobium polysaccharide and preparation method thereof ) 是由 伍曾利 于 2021-05-13 设计创作，主要内容包括：本发明涉及医药技术领域,具体涉及一种高纯度石斛活性多糖及其制备方法。该石斛多糖以新鲜饱满的石斛鲜条为原料,经清洗,晾干,打浆,提取,澄清处理,醇沉,干燥,复溶,除杂,醇沉,干燥等工艺及技术手段提取石斛多糖。石斛多糖存在于石斛鲜条中,提取过程中,经过不断的除杂工艺,不断的提高石斛多糖的纯度,本专利的石斛多糖纯度达到96.3％,石斛多糖具有增强机体免疫力、抗癌等生物活性,增强石斛多糖的增强机体免疫力及抗癌效果。(The invention relates to the technical field of medicines, and particularly relates to high-purity dendrobe active polysaccharide and a preparation method thereof. The dendrobium polysaccharide is prepared by taking fresh and plump dendrobium fresh strips as raw materials, and extracting the dendrobium polysaccharide through the processes and technical means of cleaning, airing, pulping, extracting, clarifying, alcohol precipitating, drying, redissolving, impurity removing, alcohol precipitating, drying and the like. The dendrobium polysaccharide exists in fresh dendrobium strips, and the purity of the dendrobium polysaccharide is continuously improved through a continuous impurity removal process in the extraction process, the purity of the dendrobium polysaccharide reaches 96.3%, the dendrobium polysaccharide has biological activities of enhancing the body immunity, resisting cancer and the like, and the body immunity and the anti-cancer effect of the dendrobium polysaccharide are enhanced.)

1. The preparation method of the dendrobium polysaccharide is characterized by comprising the following steps:

step 1: mixing dendrobe and water, pulping, heating to 80-100 ℃, extracting, collecting an extracting solution, cooling, centrifuging at 6000-10000 r/min for 6-12min, filtering, and collecting filtrate;

step 2: mixing the filtrate obtained in the step 1 with ethanol until the ethanol concentration of the solution is 75%, standing, centrifuging at 6000-10000 r/min for 6-12min, collecting the precipitate, and sequentially washing with 95% ethanol and acetone once respectively to obtain the washed precipitate;

and step 3: taking the washed precipitate prepared in the step 2, drying, crushing and sieving to prepare the dendrobium crude polysaccharide;

and 4, step 4: mixing the dendrobium crude polysaccharide prepared in the step 3 with water, heating to dissolve, clarifying, adding a 5% hexadecyl trimethyl ammonium bromide solution while stirring until the final concentration of the hexadecyl trimethyl ammonium bromide is 1.5%, continuously stirring, standing, centrifuging for 6-12min at 6000-10000 r/min, collecting precipitates, adding a 38% acetic acid solution at 0-10 ℃ under an ice bath condition, and continuously stirring; centrifuging at 6000-10000 rpm for 6-12min, and collecting precipitate;

and 5: mixing the precipitate prepared in the step 4 with water for dissolving, clarifying, adding 95% ethanol until the concentration of the ethanol is 60%, stirring, standing, centrifuging, taking the precipitate, sequentially washing with 95% ethanol and acetone twice respectively, and collecting the precipitate;

step 6: and (5) drying, crushing and sieving the precipitate prepared in the step (5) to obtain the dendrobium polysaccharide.

2. The preparation method according to claim 1, wherein the weight ratio of water to dendrobium in step 1 is (4-16): 1; the extraction time is 60-180 min; and reducing the temperature to 20-40 ℃.

3. The method according to claim 1 or 2, wherein the concentration of ethanol in step 2 is 95%; the standing time is 30-180 min.

4. The method according to any one of claims 1 to 3, wherein the temperature of the drying in step 3 is 70 ℃; the particle size after sieving is 60-120 meshes.

5. The preparation method according to any one of claims 1 to 4, wherein the weight ratio of the water to the crude dendrobium polysaccharide in step 4 is (10-15): 1; the heating temperature is 60 ℃; the continuous stirring time is 10 min; the standing is to stand at 20-40 ℃ for 30-180 min.

6. The method according to any one of claims 1 to 5, wherein the weight ratio of water to the precipitate in step 5 is (5-10): 1; the stirring time is 5-20 min; the standing time is 30-180 min.

7. The method according to any one of claims 1 to 6, wherein the drying temperature is 70 ℃, and the particle size after sieving is 60 to 120 mesh.

8. The method according to any one of claims 1 to 7, wherein the dendrobium comprises one or more of Dendrobium officinale, Dendrobium devonianum, Dendrobium huoshanense and Dendrobium nobile.

9. The dendrobe polysaccharide prepared by the preparation method of any one of claims 1 to 8.

10. The use of the dendrobe polysaccharide of claim 9 in the preparation of a medicament or formulation for enhancing immunity and/or cancer.

Technical Field

The invention relates to the technical field of medicines, and particularly relates to dendrobe polysaccharide and a preparation method thereof.

Background

Dendrobium officinale is a perennial epiphytic herb plant of the genus Dendrobium of the family Orchidaceae, and the area of origin is mainly distributed in Qinling and Huaihe provinces of China. Dendrobium officinale, as a well-known famous and precious traditional Chinese medicine, has the effect of 'gold in medicine' since ancient times, has the efficacies of 'yin impairment and fluid deficiency, dry and polydipsia in mouth, poor appetite, dry and retted food, asthenic fever after illness, and unclear vision', is known as the first of nine major celestial herbs in China, is highly advocated in the Yanyi of the materia medica, the compendium of the materia medica pick up the remains and the traditional Chinese herbal medicine from the New and other generations, is called as 'Qianjin weed', and is popular in the world. Modern medical research shows that the dendrobium officinale contains various chemical components such as polysaccharide, alkaloid, phenols and the like, and has the effects of protecting liver, resisting tumors, reducing blood sugar, protecting gastric ulcer, enhancing human body immunity and the like.

At present, polysaccharide extraction methods mainly comprise a heating extraction method, an enzymolysis method, an ultrasonic extraction method and the like, and the extraction methods have advantages and disadvantages respectively. Patent reports on dendrobium polysaccharide extraction mainly include in recent years: the dendrobium polysaccharide extraction method in summer (application No. 201810949604.9), the dendrobium polysaccharide separation and purification method in treentian (application No. 201110365323.7), the trewining method (application No. 201810209020.8), the dunginyuan (sugar-reducing active dendrobium polysaccharide in dunginning and extraction method thereof) (application No. 201010295717.5), and the like. The patent reports greatly enrich the extraction and purification means of the dendrobium polysaccharide, but the dendrobium polysaccharide still has defects in the aspects of extraction efficiency and polysaccharide purity. The polysaccharide extraction process in most patents includes only several conventional steps: degreasing, water extracting, alcohol precipitating, protein removing and drying; the obtained polysaccharide has a purity of 20-50% mostly without innovation.

Therefore, the high-purity dendrobe active polysaccharide and the preparation method thereof have important practical significance.

Disclosure of Invention

In view of the above, the invention provides a high-purity dendrobe active polysaccharide and a preparation method thereof, and the method can greatly improve the purity and activity of dendrobe polysaccharide and enhance the immunity and anticancer effects of dendrobe polysaccharide.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a preparation method of dendrobe polysaccharide, which comprises the following steps:

step 1: mixing dendrobe and water, pulping, heating to 80-100 ℃, extracting, collecting an extracting solution, cooling, centrifuging at 6000-10000 r/min for 6-12min, filtering, and collecting filtrate;

step 2: mixing the filtrate obtained in the step 1 with ethanol until the ethanol concentration of the solution is 75%, standing, centrifuging at 6000-10000 r/min for 6-12min, collecting the precipitate, and sequentially washing with 95% ethanol and acetone once respectively to obtain the washed precipitate;

and step 3: taking the washed precipitate prepared in the step 2, drying, crushing and sieving to prepare the dendrobium crude polysaccharide;

and 4, step 4: mixing the dendrobium crude polysaccharide prepared in the step 3 with water, heating to dissolve, clarifying, adding a 5% hexadecyl trimethyl ammonium bromide solution while stirring until the final concentration of the hexadecyl trimethyl ammonium bromide is 1.5%, continuously stirring, standing, centrifuging for 6-12min at 6000-10000 r/min, collecting precipitates, adding a 38% acetic acid solution at 0-10 ℃ under an ice bath condition, and continuously stirring; centrifuging at 6000-10000 rpm for 6-12min, and collecting precipitate;

and 5: mixing the precipitate prepared in the step 4 with water for dissolving, clarifying, adding 95% ethanol until the concentration of the ethanol is 60%, stirring, standing, centrifuging, taking the precipitate, sequentially washing with 95% ethanol and acetone twice respectively, and collecting the precipitate;

step 6: and (5) drying, crushing and sieving the precipitate prepared in the step (5) to obtain the dendrobium polysaccharide.

In some embodiments of the invention, the weight ratio of water to dendrobium in step 1 is (4-16): 1; the extraction time is 60-180 min; and reducing the temperature to 20-40 ℃.

In some embodiments of the invention, the concentration of ethanol in step 2 is 95%; the standing time is 30-180 min.

In some embodiments of the invention, the temperature of the drying in step 3 is 70 ℃; the particle size after sieving is 60-120 meshes.

In some specific embodiments of the invention, the weight ratio of the water to the dendrobium crude polysaccharide in the step 4 is (10-15): 1; the heating temperature is 60 ℃; the continuous stirring time is 10 min; the standing is to stand at 20-40 ℃ for 30-180 min.

In some embodiments of the invention, the weight ratio of water to the precipitate in step 5 is (5-10): 1; the stirring time is 5-20 min; the standing time is 30-180 min.

In some embodiments of the present invention, the drying temperature is 70 ℃, and the particle size after sieving is 60-120 meshes.

In some embodiments of the invention, the dendrobium comprises one or more of dendrobium officinale, dendrobium devonianum, dendrobium huoshanense and dendrobium nobile.

Based on the research, the invention also provides the dendrobium polysaccharide prepared by the preparation method.

More importantly, the invention also provides application of the dendrobium polysaccharide in preparing a medicine or a preparation for enhancing the immunity of the organism and/or resisting cancer.

The preparation method of the dendrobium polysaccharide provided by the invention takes fresh and plump fresh dendrobium strips as raw materials, and the dendrobium polysaccharide is extracted by the processes and technical means of cleaning, airing, pulping, extracting, clarifying, alcohol precipitating, drying, redissolving, impurity removing, alcohol precipitating, drying and the like. The dendrobium polysaccharide exists in fresh dendrobium strips, and the purity of the dendrobium polysaccharide is continuously improved through a continuous impurity removal process in the extraction process, the purity of the dendrobium polysaccharide reaches 96.3%, the dendrobium polysaccharide has biological activities of enhancing the body immunity, resisting cancer and the like, and the body immunity and the anti-cancer effect of the dendrobium polysaccharide are enhanced.

Detailed Description

The invention discloses dendrobe polysaccharide and a preparation method thereof, and can be realized by appropriately improving process parameters by taking the contents as reference by the technical personnel in the field. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.

The preparation method of the dendrobium polysaccharide provided by the invention comprises the following steps:

taking fresh dendrobium officinale strips, cleaning, adding 4-16 times of water, pulping, heating to 80-100 ℃, extracting for 60-180 min, cooling to 20-40 ℃, centrifuging for 6-12min at 6000-10000 r/min, filtering, and taking clear liquid for later use;

adding 95% ethanol into the clarified liquid to make the ethanol concentration of the solution reach 75%, standing for 30-180 min, centrifuging at 6000-10000 rpm for 6-12min, collecting precipitate, and washing with 95% ethanol and acetone respectively;

drying the precipitate at 70 ℃, crushing, and sieving with a 60-120 mesh sieve to obtain the dendrobium officinale viscous crude polysaccharide;

taking crude polysaccharide, adding 10-15 times of water, heating to 60 ℃ for dissolving, clarifying, adding a 5% hexadecyl trimethyl ammonium bromide solution under the stirring condition until the final concentration is 1.5%, continuously stirring for 10min, and standing for 30-180 min at 20-40 ℃; centrifuging at 6000-10000 r/min for 6-12min, taking the precipitate, adding a 38% acetic acid solution at 0-10 ℃ under an ice bath condition, and stirring for 10 min; centrifuging at 6000-10000 rpm for 6-12min, and taking the precipitate;

dissolving the precipitate with 5-10 times of water, clarifying, adding 95% ethanol into the solution until the ethanol concentration reaches 60%, stirring, standing for 30-180 min, centrifuging, collecting the precipitate, and washing with 95% ethanol and acetone twice respectively;

and drying the precipitate at 70 ℃, and crushing the dried precipitate through a 60-120 mesh screen to obtain the dendrobium officinale viscous high-purity polysaccharide.

The preparation method of the dendrobium polysaccharide provided by the invention takes fresh and plump fresh dendrobium strips as raw materials, and the dendrobium polysaccharide is extracted by the processes and technical means of cleaning, airing, pulping, extracting, clarifying, alcohol precipitating, drying, redissolving, impurity removing, alcohol precipitating, drying and the like. The dendrobium polysaccharide exists in fresh dendrobium strips, and the purity of the dendrobium polysaccharide is continuously improved through a continuous impurity removal process in the extraction process, the purity of the dendrobium polysaccharide reaches 96.3%, the dendrobium polysaccharide has biological activities of enhancing the body immunity, resisting cancer and the like, and the body immunity and the anti-cancer effect of the dendrobium polysaccharide are enhanced.

In the dendrobium polysaccharide and the preparation method thereof provided by the invention, the used raw materials and reagents can be purchased from the market.

The invention is further illustrated by the following examples:

example 1:

100kg of fresh dendrobium officinale strips are cleaned by tap water and dried for standby.

Adding 4 times of water, pulping with a double-pass pulping machine, heating to 80 deg.C, extracting for 60min, cooling to 20 deg.C, centrifuging at 6000 rpm for 6min, filtering, and collecting clarified liquid to obtain 275kg of liquid with solid content of 1.7%, and concentrating to obtain 10% of solid content to obtain 44.5kg of concentrated solution;

adding 95% ethanol into the concentrated solution to make the ethanol concentration of the solution reach 75%, standing for 30min, centrifuging at 6000 rpm for 6min, collecting precipitate, and washing with 95% ethanol and acetone respectively;

drying the precipitate at 70 deg.C, pulverizing, and sieving with 60 mesh sieve to obtain 3.42kg crude polysaccharide of herba Dendrobii;

taking crude polysaccharide of herba Dendrobii, adding 10 times of water, heating to 60 deg.C for dissolving, clarifying to obtain 32.3kg of liquid, adding 5% hexadecyl trimethyl ammonium bromide solution under stirring to final concentration of 1.5%, stirring for 10min, and standing at 20 deg.C for 30 min; centrifuging at 6000 rpm for 6min, collecting precipitate, adding 38% acetic acid solution at 10 deg.C under ice bath condition, and stirring for 10 min; centrifuging at 6000 rpm for 6min, and collecting precipitate;

dissolving the precipitate with 5 times of water, clarifying to obtain clarified liquid 3.8kg, adding 95% ethanol into the solution until the ethanol concentration reaches 60%, stirring, standing for 30min, centrifuging at 6000 rpm for 6min, collecting precipitate, and washing with 95% ethanol and acetone twice respectively;

drying the precipitate at 70 ℃, crushing, and sieving with a 60-mesh sieve to obtain 842g of the dendrobium officinale high-purity polysaccharide.

Example 2

100kg of fresh dendrobium officinale strips are cleaned by tap water and dried for standby.

Adding 10 times of water, pulping with a double-pass pulping machine, heating to 90 deg.C, extracting for 120min, cooling to 30 deg.C, centrifuging at 8000 rpm for 9min, filtering, collecting clarified liquid to obtain 692kg of liquid, 0.7% of solid, and concentrating to 10% of solid to obtain 43kg of concentrated solution;

adding 95% ethanol into the concentrated solution to make the ethanol concentration of the solution reach 75%, standing for 105min, centrifuging at 8000 rpm for 9min, collecting precipitate, and washing with 95% ethanol and acetone respectively;

drying the precipitate at 70 deg.C, pulverizing, and sieving with 90 mesh sieve to obtain 3.50kg crude polysaccharide of herba Dendrobii;

taking crude polysaccharide of herba Dendrobii, adding 12.5 times of water, heating to 60 deg.C for dissolving, clarifying to obtain 38.6kg of liquid, adding 5% hexadecyl trimethyl ammonium bromide solution under stirring to final concentration of 1.5%, stirring for 10min, and standing at 30 deg.C for 105 min; centrifuging at 8000 rpm for 9min, collecting precipitate, adding 38% acetic acid solution at 5 deg.C under ice bath condition, and stirring for 10 min; centrifuging at 8000 rpm for 9min, and collecting precipitate;

dissolving the precipitate with 7.5 times of water, clarifying to obtain 8.3kg of clarified liquid, adding 95% ethanol into the solution until the ethanol concentration reaches 60%, stirring, standing for 105min, centrifuging at 8000 rpm for 9min, collecting precipitate, and washing with 95% ethanol and acetone twice respectively;

and drying the precipitate at 70 ℃, crushing, and sieving with a 80-mesh sieve to obtain 840g of the dendrobium officinale high-purity polysaccharide.

Example 3

100kg of fresh dendrobium officinale strips are cleaned by tap water and dried for standby.

Adding 16 times of water, pulping with a double-pass pulping machine, heating to 100 deg.C, extracting for 180min, cooling to 40 deg.C, centrifuging at 10000 rpm for 12min, filtering, collecting clarified liquid to obtain 1280kg of liquid and 0.36% of solid, and concentrating to obtain 44.5kg of concentrated solution with solid content of 10%;

adding 95% ethanol into the concentrated solution to make the ethanol concentration of the solution reach 75%, standing for 180min, centrifuging at 10000 rpm for 12min, collecting precipitate, and washing with 95% ethanol and acetone respectively;

drying the precipitate at 70 deg.C, pulverizing, and sieving with 120 mesh sieve to obtain 3.48kg crude polysaccharide of herba Dendrobii;

taking the crude polysaccharide of herba Dendrobii, adding 15 times of water, heating to 60 deg.C for dissolving, clarifying to obtain 30.5kg of liquid, adding 5% hexadecyl trimethyl ammonium bromide solution under stirring to final concentration of 1.5%, stirring for 10min, and standing at 40 deg.C for 180 min; centrifuging at 10000 r/min for 12min, collecting precipitate, adding 38% acetic acid solution at 10 deg.C under ice bath condition, and stirring for 10 min; centrifuging at 10000 rpm for 12min, and collecting precipitate;

dissolving the precipitate with 10 times of water, clarifying to obtain 8.1kg of clarified liquid, adding 95% ethanol into the solution until the ethanol concentration reaches 60%, stirring, standing for 180min, centrifuging at 10000 rpm for 12min, collecting precipitate, and washing with 95% ethanol and acetone twice respectively;

drying the precipitate at 70 ℃, crushing, and sieving with a 120-mesh sieve to obtain 840g of the dendrobium officinale high-purity polysaccharide;

detection example:

polysaccharide detection method (phenol-sulfuric acid method): and (3) determining the content of the polysaccharide by using a sulfuric acid-phenol color development method by taking D-anhydrous glucose as a reference, wherein a standard curve equation is that y is 7.66812x +0.00000(r is 0.99926), wherein x is the mass concentration (mg/mL) of the D-anhydrous glucose and y is absorbance, and calculating the content of the polysaccharide according to a regression equation after measuring the absorbance.

Table 1: product detection

Serial number	Product(s)	Polysaccharide content
			1	The invention relates to dendrobe polysaccharide	96.3％*
2	Products of patent 201810949604.9	36.6％
			3	Products of patent 201810209020.8	43.9％
4	Products of patent 201110365323.7	46.1％

Note: compared with the comparison patent, the dendrobium polysaccharide product has obvious difference (P is less than 0.05) in statistical result.

Test example 1: dendrobe polysaccharide immunity enhancing function test

1. Test materials

1.1 test samples

The dendrobe polysaccharides prepared by the processes of the examples and the comparative examples (application numbers of 201810949604.9, 201810209020.8 and 201110365323.7) are taken as test samples.

1.2 test dose

The samples of examples and comparative examples were used in an amount of 2.4g/d, and the average amount was 40 mg/(kg. d) in terms of 60kg of adult body weight, i.e., the amount of the test sample was 40 mg/(kg. d).

In example 1 of the present invention, the test samples were set to 20, 40 and 80mg/(kg · d) for the low, medium and high 3 dose groups.

1.3 Experimental animals

Kunming mice, 18-22 g, provided by the Experimental animal center of Guangzhou Chinese medicine university, are normally bred for 3 days and then are tested.

1.4 Main instruments and reagents

UV-2501 ultraviolet-visible spectrophotometer (Shimadzu, Japan); BS110S electronic balance (Sartorius corporation); XZC-5A mouse diving tower instrument (jiangsu seons biotechnology limited); d-galactose; a SOD reagent; an MDA reagent; coomassie brilliant blue reagent; cyclophosphamide is used for injection.

2. Test method

2.1 establishment of immune hypofunction model and group administration

120 Kunming mice, male and female halves, were taken and randomly divided into a blank control group, a negative control group, example 1 high, medium and low dose groups, example 2 group, example 3 group, comparative example 1 group, comparative example 2 group and comparative example 3 group, each group containing 12 mice. Except for the blank control group, the other groups of mice were injected with cyclophosphamide injection at 50.0mg/kg in the abdominal cavity on days 1, 3 and 5. And (3) performing modeling, and simultaneously performing intragastric administration on the test sample according to the corresponding dose, performing intragastric administration on the blank control group and the negative control group for 1 time every day by using distilled water with the corresponding volume, and continuously performing intragastric administration for 14 days.

2.2 Observation index

1h after the last administration, the mice were weighed, 25% Chinese ink physiological saline solution was injected into the tail vein of 10.0mL/kg, 2 minutes and 12 minutes after the injection of the Chinese ink, 2-3 drops of blood were collected from the retroorbital venous plexus of the mice by using a capillary glass tube, and 20.0. mu.l of the blood was dissolved in 2.0mL of 0.1% Na₂CO₃In the solution, the mixture was shaken up, and the absorbance at 680nm was measured under 721 type spectrophotometry, and the phagocytosis index K was calculated according to the following formula. And (5) dissecting spleen and thymus at the same time, and calculating thymus and spleen coefficients.

The experimental data adopts a one-factor variance analysis for comparison among multiple groups, a t-test analysis method for experimental data, and differences among the groups are compared, and the data are expressed by x +/-s.

3. Results

Influence of dendrobe polysaccharide on organ weight and phagocytic function of mononuclear macrophage

Compared with a blank group, the thymus coefficient, the spleen coefficient and the carbon clearance index K of the mice in the negative control group are obviously reduced (P < 0.01); and each dosage group of the dendrobe polysaccharide test sample can obviously improve spleen coefficients and carbon particle clearance indexes (P <0.01 or 0.05) of mice with low immunity caused by cyclophosphamide, and the high and medium dosage groups can obviously improve thymus coefficients (P <0.01 or 0.05) of the mice with low immunity, so that the dendrobe polysaccharide test sample can enhance the immune function of the dendrobe polysaccharide test sample and has the effect of improving the nonspecific phagocytic function of a reticuloendothelial system of the mice. The results are shown in Table 2.

TABLE 2 Effect of high purity Dendrobii polysaccharides on immunocompromised mice (x. + -.s)

Note: compared with the blank control group, the composition of the composition,^*P<0.01; compared with the negative control group, the test results show that,^##P<0.01，^#P<0.05; in comparison with the comparative example,^※※P<0.01，^※P<0.05。

3.4 analysis

Under the experimental condition that cyclophosphamide forms immunosuppression on mice, the carbon particle clearance index of each dose group of mice of the dendrobe polysaccharide test sample is obviously higher than that of a negative control group, namely the mononuclear phagocyte function is obviously enhanced, which indicates that the dendrobium polysaccharide test sample can enhance the nonspecific immunity function of an organism. Meanwhile, the dendrobium polysaccharide test sample can increase the thymus coefficient and the spleen coefficient of a mouse.

The function of the high-purity dendrobium polysaccharide is evaluated from immune regulation, and the function of the high-purity dendrobium polysaccharide can be proved to play a role in improving the function of an immune system and the like.

Test example 2: anticancer function test of high-purity dendrobium polysaccharide

1. Test drugs:

sample preparation: high-purity dendrobe polysaccharide (examples 1-3 of the invention, comparative examples 1-3);

comparison products: cyclophosphamide.

2. Animal(s) production

NIH₆₁₅Pure mouse.

3. Tumor strain

SI₁₈₀A. EAC and L₆₁₅Mice were provided by the institute for blood, institute of Chinese medical sciences.

4. Experimental methods and results

(1) Aloe polysaccharide to mouse S₁₈₀Influence of (2)

Taking NIH pure line mice with weight of 20S 2g and dual purposes of male and female, inoculating according to the method of national anti-tumor drug in-vivo screening regulation, and inoculating S with good tumor growth in 6-9 days₁₈₀A, ascites is removed and the physiological conditions are sterilizedDiluting with saline at a ratio of l:4 to obtain tumor cell suspension (5 × 10)⁶Cells/ml). Respectively in an amount of 0.2ml_RInoculating to the right axilla of the mouse or inoculating to the abdominal cavity of the mouse.

And randomly grouping after 24h, wherein 10 of the dendrobium polysaccharide in each group are respectively administrated by intraperitoneal or subcutaneous injection once a day, the dose of the dendrobium polysaccharide is 60mg/kg, the dose of the cyclophosphamide is 60mg/kg, the blank control group is administrated with physiological saline with the same volume for 10d continuously, the mice inoculated under the skin are killed the next day after the medicine is stopped, tumor masses are taken out, the weight is measured, and the tumor inhibition rate is calculated. The number of days of life was observed after the mice inoculated with the abdominal cavity had stopped taking the drug, and the life extension rate was calculated. The results are shown in the following table.

Table 3: dendrobe polysaccharide pair mouse S₁₈₀Influence of (2)

Note: compared with the blank control group (physiological saline group),^*P<0.01; in comparison with the comparative example group,^#P<0.01。

table 4: effect of dendrobe polysaccharide on mouse S180A

Note: compared with the physiological saline (blank control group),^*P<0.01; in comparison with the comparative example group,^#P<0.01。

as can be seen from the test results, the dendrobii polysaccharide has significant effect on resisting mouse S₁₈₀Action of (1) against solid tumors S₁₈₀Or ascites type S₁₈₀(S₁₈₀A) Has strong antagonistic effect, and the effect is remarkably superior to that of a comparative product.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.