CN115286721A - Active polysaccharide, active polysaccharide composition and application of active polysaccharide composition in preparation of product with effect of preventing or treating gastric injury - Google Patents
Active polysaccharide, active polysaccharide composition and application of active polysaccharide composition in preparation of product with effect of preventing or treating gastric injury Download PDFInfo
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Abstract
The invention relates to the technical field of biological medicines, and particularly discloses active polysaccharide, an active polysaccharide composition and application of the active polysaccharide composition in preparation of a product with a function of preventing or treating gastric injury. The active polysaccharide is the codonopsis pilosula polysaccharide prepared by the brand new method; the active polysaccharide composition comprises the codonopsis pilosula polysaccharide and the glycyrrhiza polysaccharide. The research shows that: compared with crude codonopsis pilosula polysaccharide, the codonopsis pilosula polysaccharide and active polysaccharide composition prepared by the method provided by the invention have the advantages that the gastric injury preventing or treating effect is remarkably improved, and the codonopsis pilosula polysaccharide and active polysaccharide composition has an excellent gastric injury preventing or treating effect. Therefore, the active polysaccharide and the active polysaccharide composition have important application values when being used as effective components for developing medicaments, foods or dietary supplements with the effect of preventing or treating the gastric injury and developing stomach-protecting medicaments.
Description
Technical Field
The invention relates to the technical field of biomedicine, and in particular relates to active polysaccharide, an active polysaccharide composition and application thereof in preparation of a product with a function of preventing or treating gastric injury.
Background
The great pressure of life and work makes drinking gradually become a habit of people, and a plurality of researches show that acute or chronic injury of gastric mucosa is closely related to long-term large-scale and unopposed drinking, and the acute or chronic injury of gastric mucosa is mainly manifested as congestion, erosion, ulcer and the like of gastric mucosa.
At present, the treatment of gastric ulcer and alcoholic gastric injury is mainly carried out by using chemical drugs or traditional Chinese medicines. Chemical drugs include H2 proton pump inhibitors, M1 receptor blockers, etc., but long-term use of chemical drugs can have significant side effects, such as: arrhythmia, sexual dysfunction, etc. Compared with chemical drugs, the traditional Chinese medicine has the advantages of small toxic and side effects and multiple targets.
The active polysaccharide extracted from the traditional Chinese medicine has various physiological activities, and has a certain protection effect on alcoholic stomach injury. Although the codonopsis pilosula polysaccharide has been reported to have the effect of preventing or treating gastric injury in the prior art, the effect of the codonopsis pilosula polysaccharide prepared by the prior art on preventing or treating gastric injury needs to be further improved.
Therefore, it is of great value to develop an active polysaccharide that has a better effect of preventing or treating gastric injury.
Disclosure of Invention
In order to solve at least one technical problem in the prior art, the invention firstly provides an active polysaccharide.
The technical scheme of the invention is as follows:
the invention firstly provides an active polysaccharide, wherein the active polysaccharide is codonopsis pilosula polysaccharide, and the codonopsis pilosula polysaccharide is prepared by a method comprising the following steps:
s11, taking traditional Chinese medicine codonopsis pilosula, and preparing codonopsis pilosula crude polysaccharide by using a water extraction and alcohol precipitation method;
s12, adding water to dissolve the crude codonopsis pilosula polysaccharide, dialyzing by using a dialysis bag with the aperture of 18-22 kD to obtain first dialysate and first cut-off liquid, and concentrating and drying the first cut-off liquid to obtain first codonopsis pilosula polysaccharide;
s13, dialyzing the first dialysate by using a dialysis bag with the density of 8-12 kD to obtain a second dialysate and a second trapped fluid, and concentrating and drying the second trapped fluid to obtain second codonopsis pilosula polysaccharide;
s14, dialyzing the second dialysate by using a dialysis bag with the concentration of 2-4 kD to obtain a third dialysate and a third interception liquid, and concentrating and drying the third interception liquid to obtain a third codonopsis pilosula polysaccharide; concentrating and drying the third dialysate to obtain a fourth codonopsis pilosula polysaccharide; and taking the third codonopsis pilosula polysaccharide to obtain the codonopsis pilosula polysaccharide.
The inventor finds that the codonopsis pilosula polysaccharide prepared by different methods has great difference in the effects of preventing or treating gastric injury. The inventors have surprisingly found in a number of experiments that: the third codonopsis pilosula polysaccharide prepared by the method has excellent effect of preventing or treating gastric injury; the effect of preventing or treating gastric injury is obviously higher than that of the crude polysaccharide of the codonopsis pilosula; meanwhile, the content of the polysaccharide is obviously higher than that of other codonopsis pilosula polysaccharides obtained in the preparation process.
Preferably, in step S12, dialysis is performed with a dialysis bag having a pore size of 20 kD.
Preferably, S13 is dialyzed with a dialysis bag having a pore size of 10 kD.
Preferably, dialysis is performed in step S14 using a dialysis bag having a pore size of 3 kD.
Preferably, the specific method for preparing crude codonopsis pilosula polysaccharide by using a water extraction and alcohol precipitation method in step S11 comprises the following steps:
extracting radix Codonopsis with water under heating and refluxing to obtain radix Codonopsis extractive solution; then, concentrating the codonopsis pilosula extracting solution to obtain a codonopsis pilosula concentrated solution, and adding ethanol into the codonopsis pilosula concentrated solution to ensure that the final volume fraction of the ethanol in the codonopsis pilosula concentrated solution is 60-80%; and finally, standing and taking the precipitate to obtain the codonopsis pilosula crude polysaccharide.
Preferably, the using ratio of the codonopsis pilosula to the water is 1g:6 to 12mL.
Most preferably, the amount ratio of the process water is 1g:8mL.
Preferably, the codonopsis pilosula extracting solution is concentrated to obtain a codonopsis pilosula concentrated solution, so that the volume of the codonopsis pilosula concentrated solution is 1/3-1/5 of the volume of water.
Most preferably, the Codonopsis pilosula extract is concentrated to give a Codonopsis pilosula concentrate such that the volume of the Codonopsis pilosula concentrate is 1/4 of the volume of water.
Preferably, the final volume fraction of ethanol in the codonopsis pilosula concentrated solution is 70%.
The invention also provides an active polysaccharide composition, which comprises codonopsis pilosula polysaccharide and glycyrrhiza polysaccharide;
the codonopsis pilosula polysaccharide is prepared according to the method.
Preferably, the weight ratio of the codonopsis pilosula polysaccharide to the glycyrrhiza polysaccharide is 2-4:1.
Preferably, the glycyrrhiza polysaccharide is prepared by a method comprising the following steps:
s21, taking traditional Chinese medicine liquorice, and preparing liquorice crude polysaccharide by using a water extraction and alcohol precipitation method;
s22, dissolving crude glycyrrhiza polysaccharide in water, dialyzing by using a dialysis bag with the aperture of 14-16 kD to obtain first dialysate and first cut-off liquid, and concentrating and drying the first cut-off liquid to obtain first glycyrrhiza polysaccharide;
s23, dialyzing the first dialysate by using a dialysis bag of 7-9 kD to obtain a second dialysate and a second trapped fluid, and concentrating and drying the second trapped fluid to obtain second glycyrrhiza polysaccharide; concentrating and drying the second dialysate to obtain third glycyrrhiza polysaccharide; and taking the second glycyrrhiza polysaccharide to obtain the glycyrrhiza polysaccharide.
Preferably, in step S22, dialysis is performed with a dialysis bag having a pore size of 15 kD.
Preferably, dialysis is performed in step S23 using a dialysis bag having a pore size of 8 kD.
Preferably, the specific method for preparing the crude licorice polysaccharide by using the water extraction and alcohol precipitation method in step S21 is as follows:
heating and refluxing traditional Chinese medicine liquorice with water to obtain a liquorice extracting solution; then, concentrating the licorice extract to obtain a licorice concentrated solution, and adding ethanol into the licorice concentrated solution to ensure that the final volume fraction of the ethanol in the licorice concentrated solution is 60-80%; and finally, standing and taking the precipitate to obtain the licorice crude polysaccharide.
Preferably, the dosage ratio of the liquorice to the water is 1g:6 to 12mL.
Most preferably, the ratio of the amount of licorice to water is 1g:8mL.
Preferably, the licorice extract is concentrated to obtain a licorice concentrate such that the volume of the licorice concentrate is 1/3 to 1/5 of the volume of water.
Most preferably, the licorice extract is concentrated to obtain a licorice concentrate such that the volume of the licorice concentrate is 1/4 of the volume of water.
Preferably, the final volume fraction of ethanol in the licorice concentrate is made to be 70%.
The inventor surprisingly found in further research; compared with the third codonopsis pilosula polysaccharide, the stomach injury preventing or treating effect of the active polysaccharide composition obtained by combining the third codonopsis pilosula polysaccharide prepared by the method disclosed by the invention and the second glycyrrhiza polysaccharide prepared by the method disclosed by the invention is further greatly improved, and the active polysaccharide composition is far higher than that of codonopsis pilosula crude polysaccharide. However, the inventors have also found in their studies that the active polysaccharide composition obtained by combining the third codonopsis pilosula polysaccharide prepared according to the above-mentioned method of the present invention with other polysaccharides, or the active polysaccharide composition obtained by combining the third codonopsis pilosula polysaccharide with other glycyrrhiza polysaccharides prepared according to the method of the present invention, does not further improve the gastric lesion preventing or treating effect of the third codonopsis pilosula polysaccharide.
The invention also provides application of the active polysaccharide or the active polysaccharide composition in preparing a product with the effect of preventing or treating gastric injury.
Preferably, the gastric injury is acute alcoholic gastric injury.
Preferably, the product is a medicament, food or dietary supplement.
The invention also provides the application of the active polysaccharide or the active polysaccharide composition in preparing a stomach-protecting medicine.
Has the advantages that: the invention provides a brand new codonopsis pilosula polysaccharide; compared with crude codonopsis pilosula polysaccharide, the codonopsis pilosula polysaccharide prepared by the method provided by the invention has the advantages that the gastric injury preventing or treating effect is remarkably improved, and the codonopsis pilosula polysaccharide has an excellent gastric injury preventing or treating effect. After the second glycyrrhiza polysaccharide prepared by the method is further combined with the third codonopsis pilosula polysaccharide (namely the codonopsis pilosula polysaccharide) prepared by the method, the effect of preventing or treating gastric injury is further greatly improved; the effect of treating gastric injury is far higher than that of the crude polysaccharide of codonopsis pilosula. The codonopsis pilosula polysaccharide and active polysaccharide composition has excellent effects of preventing or treating gastric injury; therefore, the compound has important application value when being used as an effective component for developing medicaments, foods or dietary supplements with the effect of preventing or treating gastric injury and developing stomach-protecting medicaments.
Detailed Description
The technical solution of the present invention will be clearly and completely described with reference to the following examples. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
EXAMPLE 1 preparation of Codonopsis pilosula polysaccharide
S11, taking 500g of traditional Chinese medicine codonopsis pilosula, adding 4L of water, heating and refluxing for extraction for 1.5 hours at 100 ℃ to obtain a codonopsis pilosula extracting solution, and concentrating the codonopsis pilosula extracting solution to 1L to obtain a codonopsis pilosula concentrated solution; adding 95% ethanol by volume fraction into radix Codonopsis concentrated solution to make ethanol in radix Codonopsis concentrated solution at final volume fraction of 70%, standing for 6 hr, filtering, and collecting solid to obtain radix Codonopsis crude polysaccharide;
s12, adding water to dissolve crude codonopsis pilosula polysaccharides, dialyzing by using a dialysis bag with the aperture of 20kD to obtain first dialysate and first cut-off liquid, and concentrating and drying the first cut-off liquid to obtain first codonopsis pilosula polysaccharides;
s13, dialyzing the first dialysate by using a 10kD dialysis bag to obtain a second dialysate and a second trapped fluid, and concentrating and drying the second trapped fluid to obtain a second codonopsis pilosula polysaccharide;
s14, dialyzing the second dialysate by using a 3kD dialysis bag to obtain a third dialysate and a third interception liquid, and concentrating and drying the third interception liquid to obtain a third codonopsis pilosula polysaccharide; concentrating and drying the third dialysate to obtain a fourth codonopsis pilosula polysaccharide;
and taking the third codonopsis pilosula polysaccharide to obtain the codonopsis pilosula polysaccharide.
EXAMPLE 2 preparation of Glycyrrhiza polysaccharide
S21, taking 500g of traditional Chinese medicine liquorice, adding 4L of water, heating and refluxing for 1.5 hours at 100 ℃ to obtain a liquorice extracting solution, and concentrating the liquorice extracting solution to 1L to obtain a liquorice concentrated solution; adding 95% ethanol to the Glycyrrhrizae radix concentrated solution to make the final volume fraction of ethanol in the Glycyrrhrizae radix concentrated solution 70%, standing for 6 hr, filtering, and collecting solid to obtain Glycyrrhrizae radix crude polysaccharide;
s22, adding water to dissolve the crude licorice polysaccharide, dialyzing by using a dialysis bag with the aperture of 15kD to obtain first dialysate and first cut-off liquid, and concentrating and drying the first cut-off liquid to obtain first licorice polysaccharide;
s23, dialyzing the first dialysate by using a dialysis bag with the density of 8kD to obtain a second dialysate and a second trapped fluid, and concentrating and drying the second trapped fluid to obtain second glycyrrhiza polysaccharide; concentrating and drying the second dialysate to obtain third glycyrrhiza polysaccharide;
and taking the second glycyrrhiza polysaccharide to obtain the glycyrrhiza polysaccharide.
EXAMPLE 3 preparation of active polysaccharide composition
The third codonopsis pilosula polysaccharide prepared according to the method in the embodiment 1 and the second glycyrrhiza polysaccharide prepared according to the method in the embodiment 2 are uniformly mixed according to the weight ratio of 3:1 to obtain the active polysaccharide composition.
Comparative example 1 preparation of active polysaccharide composition
The third codonopsis pilosula polysaccharide prepared according to the method in the embodiment 1 and the crude licorice polysaccharide prepared according to the method in the embodiment 2 are uniformly mixed according to the weight ratio of 3:1 to obtain the active polysaccharide composition.
Comparative example 2 preparation of active polysaccharide composition
The third codonopsis pilosula polysaccharide prepared according to the method in the embodiment 1 and the first glycyrrhiza polysaccharide prepared according to the method in the embodiment 2 are uniformly mixed according to the weight ratio of 3:1 to obtain the active polysaccharide composition.
Comparative example 3 preparation of an active polysaccharide composition
The third codonopsis pilosula polysaccharide prepared according to the method in the embodiment 1 and the third glycyrrhiza polysaccharide prepared according to the method in the embodiment 2 are uniformly mixed according to the weight ratio of 3:1 to obtain the active polysaccharide composition.
Examples of the experiments
88 balb/c mice with the weight of 20 +/-2 g are taken, the male and female mice are 44 respectively, the male and female mice are randomly divided into 11 groups, each group comprises 8 mice, and the groups respectively comprise a blank group, a model group and an experimental group. Blank and model groups were given equal volumes of 0.75% NaCl solution; respectively administering 0.75% NaCl solution containing active polysaccharide or active polysaccharide composition to the model group, wherein the administration dose of the active polysaccharide or the active polysaccharide composition is 5mg/kg; the administration is performed by gavage once a day for 7 days.
1h after the last administration, each group of mice was gavaged with 300. Mu.L of 50% ethanol. After 2h ethanol injury, the mice were sacrificed, dissected, the whole stomach was taken out, washed with physiological saline, immersed in 2% formalin solution for fixation for 10min, and then the gastric ulcer area was measured under a microscope, and the sum of the ulcer areas was the ulcer index (mm) 2 ) The specific results are shown in Table 1. Wherein, the smaller the ulcer index, the better the effect of preventing or treating gastric injury.
Wherein the active polysaccharide or active polysaccharide composition administered to the experimental group is: the third codonopsis polysaccharide (i.e. the codonopsis polysaccharide of the present invention), the crude codonopsis polysaccharide, the first codonopsis polysaccharide, the second codonopsis polysaccharide and the fourth codonopsis polysaccharide prepared according to the method of example 1, the active polysaccharide composition prepared in example 3, the active polysaccharide composition prepared in comparative example 1, the active polysaccharide composition prepared in comparative example 2 and the active polysaccharide composition prepared in comparative example 3.
TABLE 1 protective action of active polysaccharides or active polysaccharide combinations against acute gastric injury caused by ethanol
In comparison to the blank set, the results, ## p is less than 0.01, compared with the model group, ** p＜0.01， * p＜0.05。
as can be seen from Table 1, after the mice are infused with gastric alcohol, the gastric ulcer index of the mice is obviously increased, which indicates that acute drinking has great damage to the mice stomach.
As can be seen from the data in table 1, the index differences of the third codonopsis pilosula polysaccharide (i.e., the codonopsis pilosula polysaccharide of the present invention), the crude codonopsis pilosula polysaccharide, the first codonopsis pilosula polysaccharide, the second codonopsis pilosula polysaccharide, and the fourth codonopsis pilosula polysaccharide, which are prepared according to the method of example 1, to the gastric ulcer of mice are great; wherein the index of gastric ulcer of mice by the third codonopsis pilosula polysaccharide (namely the codonopsis pilosula polysaccharide of the invention) prepared according to the method in the embodiment 1 is obviously lower than that of the crude codonopsis pilosula polysaccharide, and is also obviously lower than that of the first codonopsis pilosula polysaccharide, the second codonopsis pilosula polysaccharide and the fourth codonopsis pilosula polysaccharide. In addition, as can be seen from the data in table 1, the first codonopsis pilosula polysaccharide has a higher gastric ulcer index in mice than the crude codonopsis pilosula polysaccharide. This indicates that: the codonopsis pilosula polysaccharide prepared by different methods has great difference in the effect of preventing or treating gastric injury; however, only the third codonopsis pilosula polysaccharide prepared by the method of the present invention has more excellent effects of preventing or treating gastric injury; compared with the crude polysaccharide of radix codonopsitis, the effect of preventing or treating gastric injury can be obviously improved; compared with crude codonopsis pilosula polysaccharide, the codonopsis pilosula polysaccharide obtained in the preparation process of the invention can not remarkably improve the effect of preventing or treating gastric injury, and even can be reduced.
As can be seen from the data in Table 1, the active polysaccharide composition prepared in example 3 has a gastric ulcer index in mice substantially lower than that of the third Codonopsis polysaccharide (i.e., the Codonopsis polysaccharide of the present invention) prepared in example 1, and is also much lower than that of the crude Codonopsis polysaccharide; however, the active polysaccharide compositions prepared in comparative examples 1 to 3 had gastric ulcer index in mice that was not reduced as compared to the third Codonopsis polysaccharide (i.e., the Codonopsis polysaccharide of the present invention) prepared according to the method of example 1. This indicates that: only the active polysaccharide composition obtained by combining the third codonopsis pilosula polysaccharide prepared by the method of the present invention with the second glycyrrhiza polysaccharide prepared by the method of the present invention has the effect of preventing or treating gastric injury, which is further greatly improved compared with the third codonopsis pilosula polysaccharide. However, the active polysaccharide composition obtained by combining the third codonopsis pilosula polysaccharide prepared according to the above-mentioned method of the present invention with other glycyrrhiza polysaccharides prepared according to the method of the present invention does not further improve the gastric lesion preventing or treating effect of the third codonopsis pilosula polysaccharide.
Claims (10)
1. The active polysaccharide is characterized by being codonopsis pilosula polysaccharide, and being prepared by a method comprising the following steps:
s11, taking traditional Chinese medicine codonopsis pilosula, and preparing codonopsis pilosula crude polysaccharide by using a water extraction and alcohol precipitation method;
s12, adding water to dissolve crude codonopsis pilosula polysaccharides, dialyzing by using a dialysis bag with the aperture of 18-22 kD to obtain first dialysate and first cut-off liquid, and concentrating and drying the first cut-off liquid to obtain first codonopsis pilosula polysaccharides;
s13, dialyzing the first dialysate by using a dialysis bag with the density of 8-12 kD to obtain a second dialysate and a second trapped fluid, and concentrating and drying the second trapped fluid to obtain second codonopsis pilosula polysaccharide;
s14, dialyzing the second dialysate by using a dialysis bag with the concentration of 2-4 kD to obtain a third dialysate and a third interception liquid, and concentrating and drying the third interception liquid to obtain a third codonopsis pilosula polysaccharide; concentrating and drying the third dialysate to obtain a fourth codonopsis pilosula polysaccharide; and taking the third codonopsis pilosula polysaccharide to obtain the codonopsis pilosula polysaccharide.
2. The active polysaccharide according to claim 1, wherein dialysis is carried out in step S12 using a dialysis bag having a pore size of 20 kD.
3. The active polysaccharide according to claim 1, wherein S13 is dialyzed with a dialysis bag having a pore size of 10 kD.
4. The active polysaccharide according to claim 1, wherein dialysis is carried out in step S14 using a dialysis bag having a pore size of 3 kD.
5. An active polysaccharide composition, wherein the active polysaccharide composition comprises codonopsis pilosula polysaccharide and glycyrrhiza polysaccharide;
the codonopsis pilosula polysaccharide is prepared by the method of any one of claims 1 to 4.
6. The active polysaccharide composition of claim 5, wherein the glycyrrhiza polysaccharide is prepared by a process comprising the steps of:
s21, taking traditional Chinese medicine liquorice, and preparing liquorice crude polysaccharide by using a water extraction and alcohol precipitation method;
s22, adding water to dissolve the crude licorice polysaccharide, dialyzing by using a dialysis bag with the aperture of 14-16 kD to obtain first dialysate and first cut-off liquid, and concentrating and drying the first cut-off liquid to obtain first licorice polysaccharide;
s23, dialyzing the first dialysate by using a 7-9 kD dialysis bag to obtain a second dialysate and a second trapped fluid, and concentrating and drying the second trapped fluid to obtain a second glycyrrhiza polysaccharide; concentrating and drying the second dialysate to obtain third glycyrrhiza polysaccharide; and taking the second glycyrrhiza polysaccharide to obtain the glycyrrhiza polysaccharide.
7. The active polysaccharide composition of claim 6, wherein in step S22 dialysis is performed with a dialysis bag having a pore size of 15 kD.
8. The active polysaccharide composition of claim 6, wherein in step S23 dialysis is performed with a dialysis bag having a pore size of 8 kD.
9. Use of an active polysaccharide or active polysaccharide composition according to any one of claims 1 to 8 in the manufacture of a product for the prevention or treatment of gastric injury.
10. Use of an active polysaccharide or active polysaccharide composition as claimed in any one of claims 1 to 8 in the manufacture of a stomach care medicament.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1648136A (en) * | 2005-01-12 | 2005-08-03 | 贵州省中国科学院天然产物化学重点实验室 | Companumoea root polyose, derivative and its preparing method and use |
| CN104147042A (en) * | 2014-09-02 | 2014-11-19 | 遵义医学院 | Application of sulfated codonopsis pilosul polysaccharide to preparation of herpes simplex virus type I resistance medicament and preparation method of herpes simplex virus type I resistance medicament |
| CN106188326A (en) * | 2016-07-15 | 2016-12-07 | 山东省食品药品检验研究院 | The extracting method of aging ingredient and application in Radix Codonopsis |
| CN106632710A (en) * | 2016-10-12 | 2017-05-10 | 山西医科大学 | Radix codonopsis homogeneous polysaccharide with anti-gastric ulcer effect and preparation method and application thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1648136A (en) * | 2005-01-12 | 2005-08-03 | 贵州省中国科学院天然产物化学重点实验室 | Companumoea root polyose, derivative and its preparing method and use |
| CN104147042A (en) * | 2014-09-02 | 2014-11-19 | 遵义医学院 | Application of sulfated codonopsis pilosul polysaccharide to preparation of herpes simplex virus type I resistance medicament and preparation method of herpes simplex virus type I resistance medicament |
| CN106188326A (en) * | 2016-07-15 | 2016-12-07 | 山东省食品药品检验研究院 | The extracting method of aging ingredient and application in Radix Codonopsis |
| CN106632710A (en) * | 2016-10-12 | 2017-05-10 | 山西医科大学 | Radix codonopsis homogeneous polysaccharide with anti-gastric ulcer effect and preparation method and application thereof |
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