CN115286721B - Active polysaccharide, active polysaccharide composition and application thereof in preparation of products with effect of preventing or treating gastric injury - Google Patents
Active polysaccharide, active polysaccharide composition and application thereof in preparation of products 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 an active polysaccharide, an active polysaccharide composition and application thereof in preparing a product with a stomach injury prevention or treatment effect. The active polysaccharide is pilose asiabell root polysaccharide prepared by adopting the brand new method; the active polysaccharide composition comprises the codonopsis pilosula polysaccharide and the liquorice polysaccharide. Studies have shown that: compared with the crude pilose asiabell root polysaccharide, the pilose asiabell root polysaccharide and the active polysaccharide composition prepared by the method have the advantages that the effect of preventing or treating gastric injury is obviously improved, and the pilose asiabell root polysaccharide and the active polysaccharide composition have excellent effect of preventing or treating gastric injury. Therefore, the active polysaccharide and the active polysaccharide composition provided by the invention are used as active ingredients for developing medicaments, foods or dietary supplements with the effect of preventing or treating gastric injury, and have important application values for developing stomach protecting medicaments.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to an active polysaccharide, an active polysaccharide composition and application thereof in preparing products with the effect of preventing or treating gastric injury.
Background
The great pressure of life and work makes drinking become one habit of people, and many researches show that acute or chronic damage of gastric mucosa is closely related to long-term drinking without restriction, and the acute or chronic damage of gastric mucosa is mainly manifested by congestion, erosion, ulcer and the like of gastric mucosa.
At present, the gastric ulcer and alcoholic gastric injury are mainly treated by chemical medicines or traditional Chinese medicines. Chemical drugs include H2 proton pump inhibitors, M1 receptor blockers, etc., but obvious side effects occur with long-term use of chemical drugs, such as: arrhythmia, sexual dysfunction, and the like. Compared with chemical medicines, 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 gastric injury. Although the dangshen polysaccharide has the effect of preventing or treating gastric injury in the prior art, the effect of preventing or treating gastric injury of the dangshen polysaccharide prepared by the prior art is still to be further improved.
Therefore, the development of an active polysaccharide with better prevention or treatment effect on gastric injury has important application value.
Disclosure of Invention
In order to solve at least one technical problem existing 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 then preparing codonopsis pilosula crude polysaccharide by using a water extraction and alcohol precipitation method;
s12, dissolving the crude codonopsis pilosula polysaccharide in water, dialyzing by using a dialysis bag with the aperture of 18-22 kD to obtain a first dialysate and a first trapped fluid, concentrating and drying the first trapped fluid to obtain the first codonopsis pilosula polysaccharide;
s13, dialyzing the first dialysate by using a dialysis bag with the size of 8-12 kD to obtain second dialysate and second trapped fluid, concentrating and drying the second trapped fluid to obtain second codonopsis pilosula polysaccharide;
s14, dialyzing the second dialyzate by using a dialysis bag with a molecular weight of 2-4 kD to obtain a third dialyzate and a third trapped fluid, concentrating and drying the third trapped fluid to obtain third dangshen polysaccharide; concentrating and drying the third dialysate to obtain fourth codonopsis pilosula polysaccharide; and taking the third dangshen polysaccharide to obtain the dangshen polysaccharide.
The inventors found in the study that the effect of the Codonopsis pilosula polysaccharide prepared by different methods for preventing or treating gastric injury is greatly different. 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 pilose asiabell root crude polysaccharide; and is also obviously higher than other pilose asiabell root polysaccharide obtained in the preparation process of the invention.
Preferably, in step S12, the dialysis is performed with a dialysis bag having a pore size of 20 kD.
Preferably, the dialysis is performed in S13 using a dialysis bag with a pore size of 10 kD.
Preferably, in step S14, the dialysis is performed with a dialysis bag having a pore size of 3 kD.
Preferably, the specific method for preparing the codonopsis pilosula crude polysaccharide by using the water extraction and alcohol precipitation method in the step S11 is as follows:
reflux-extracting radix Codonopsis with water under heating to obtain radix Codonopsis extractive solution; concentrating the radix codonopsis extract to obtain radix codonopsis concentrate, and adding ethanol into the radix codonopsis concentrate to make the final volume fraction of ethanol in the radix codonopsis concentrate be 60-80%; finally, standing and taking out the precipitate to obtain the codonopsis pilosula crude polysaccharide.
Preferably, the dosage ratio of the party participating in water is 1g: 6-12 mL.
Most preferably, the dosage ratio of party-participating water is 1g:8mL.
Preferably, the radix codonopsis extract is concentrated to obtain radix codonopsis concentrate, so that the volume of the radix codonopsis concentrate is 1/3-1/5 of the volume of water.
Most preferably, the radix Codonopsis extract is concentrated to give a radix Codonopsis concentrate such that the volume of the radix Codonopsis concentrate is 1/4 of the volume of water.
Preferably, the final volume fraction of ethanol in the codonopsis pilosula concentrate is made to be 70%.
The invention also provides an active polysaccharide composition, which comprises codonopsis pilosula polysaccharide and liquorice polysaccharide;
the codonopsis pilosula polysaccharide is prepared by the method.
Preferably, the weight ratio of the codonopsis pilosula polysaccharide to the liquorice polysaccharide is 2-4:1.
Preferably, the glycyrrhiza polysaccharide is prepared by a method comprising the following steps:
s21, preparing licorice serving as a traditional Chinese medicine, and then preparing crude polysaccharide of the licorice by using a water extraction and alcohol precipitation method;
s22, adding water into the crude licorice polysaccharide to dissolve, then dialyzing by using a dialysis bag with the aperture of 14-16 kD to obtain a first dialysate and a first trapped fluid, concentrating and drying the first trapped fluid to obtain first licorice polysaccharide;
s23, dialyzing the first dialysate by using a 7-9 kD dialysis bag to obtain second dialysate and second trapped fluid, concentrating and drying the second trapped fluid to obtain second glycyrrhiza polysaccharide; concentrating and drying the second dialyzate to obtain third glycyrrhiza polysaccharide; and taking the second glycyrrhiza polysaccharide to obtain the glycyrrhiza polysaccharide.
Preferably, in step S22, the dialysis is performed with a dialysis bag having a pore size of 15 kD.
Preferably, in step S23, dialysis is performed with a dialysis bag having a pore size of 8 kD.
Preferably, the specific method for preparing the licorice crude polysaccharide by water extraction and alcohol precipitation in the step S21 is as follows:
heating and reflux extracting Glycyrrhrizae radix with water to obtain Glycyrrhrizae radix extractive solution; concentrating the licorice extractive solution to obtain licorice concentrate, and adding ethanol to the licorice concentrate to obtain ethanol with final volume fraction of 60-80%; finally, standing and taking out the precipitate to obtain the licorice crude polysaccharide.
Preferably, the dosage ratio of liquorice to water is 1g: 6-12 mL.
Most preferably, the dosage ratio 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 70%.
The inventors have surprisingly found in further studies; the effect of preventing or treating gastric injury of the active polysaccharide composition obtained by combining the third dangshen polysaccharide prepared by the method and the second licorice polysaccharide prepared by the method is further greatly improved compared with that of the third dangshen polysaccharide, and the effect is far higher than that of the dangshen crude polysaccharide. However, the inventors have also found in the study that the active polysaccharide composition obtained by combining the third dangshen polysaccharide prepared by the above-described method of the present invention with other polysaccharides, or the active polysaccharide composition obtained by combining the third dangshen polysaccharide prepared by the method of the present invention with other licorice polysaccharides, does not further improve the stomach injury preventing or treating effect of the third dangshen polysaccharide.
The invention also provides application of the active polysaccharide or the active polysaccharide composition in preparing products with the effect of preventing or treating gastric injury.
Preferably, the gastric injury is acute alcoholic gastric injury.
Preferably, the product is a pharmaceutical, food or dietary supplement.
The invention also provides application of the active polysaccharide or the active polysaccharide composition in preparing stomach protecting medicines.
The beneficial effects are that: the invention provides a brand new codonopsis pilosula polysaccharide; compared with the crude pilose asiabell root polysaccharide, the pilose asiabell root polysaccharide prepared by the method has the advantages that the effect of preventing or treating gastric injury is obviously improved, and the pilose asiabell root polysaccharide has excellent effect of preventing or treating gastric injury. After the second licorice polysaccharide prepared by the method is further combined with the third dangshen polysaccharide prepared by the method (namely the dangshen polysaccharide) provided by the invention, 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 pilose asiabell root crude polysaccharide. The codonopsis pilosula polysaccharide and active polysaccharide composition provided by the invention have the advantages of excellent prevention or treatment of gastric injury; therefore, the active ingredient of the composition is used for developing medicines, foods or dietary supplements with the effect of preventing or treating gastric injury, and has important application value for developing stomach protecting medicines.
Detailed Description
The technical scheme of the present invention will be clearly and completely described in the following examples. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
EXAMPLE 1 preparation of Codonopsis pilosula polysaccharide
S11, taking 500g of traditional Chinese medicine codonopsis pilosula, adding 4L of water, heating and reflux-extracting for 1.5h at 100 ℃ to obtain a codonopsis pilosula extract, and concentrating the codonopsis pilosula extract to 1L to obtain a codonopsis pilosula concentrated solution; adding 95% ethanol into the concentrated solution of radix Codonopsis to obtain final volume fraction of ethanol in the concentrated solution of radix Codonopsis of 70%, standing for 6 hr, filtering, and collecting solid to obtain crude polysaccharide of radix Codonopsis;
s12, dissolving the crude codonopsis pilosula polysaccharide in water, dialyzing by using a dialysis bag with the aperture of 20kD to obtain a first dialysate and a first trapped fluid, concentrating and drying the first trapped fluid to obtain the first codonopsis pilosula polysaccharide;
s13, dialyzing the first dialysate by using a 10kD dialysis bag to obtain a second dialysate and a second trapped fluid, concentrating and drying the second trapped fluid to obtain second codonopsis pilosula polysaccharide;
s14, dialyzing the second dialyzate by using a 3kD dialysis bag to obtain a third dialyzate and a third trapped fluid, concentrating and drying the third trapped fluid to obtain third dangshen polysaccharide; concentrating and drying the third dialysate to obtain fourth codonopsis pilosula polysaccharide;
taking the third dangshen polysaccharide to obtain the dangshen polysaccharide.
EXAMPLE 2 preparation of Glycyrrhiza polysaccharide
S21, taking 500g of Chinese medicinal licorice, then adding 4L of water, heating and reflux-extracting for 1.5h at 100 ℃ to obtain licorice extract, and concentrating the licorice extract to 1L to obtain licorice concentrate; adding 95% ethanol into the concentrated solution of Glycyrrhrizae radix to obtain final volume fraction of ethanol in the concentrated solution of Glycyrrhrizae radix of 70%, standing for 6 hr, filtering, and collecting solid to obtain Glycyrrhrizae radix crude polysaccharide;
s22, dissolving the crude licorice polysaccharide in water, dialyzing with a dialysis bag with the aperture of 15kD to obtain a first dialysate and a first trapped fluid, concentrating and drying the first trapped fluid to obtain a first licorice polysaccharide;
s23, dialyzing the first dialysate by using a 8kD dialysis bag to obtain a second dialysate and a second trapped fluid, concentrating and drying the second trapped fluid to obtain second glycyrrhiza polysaccharide; concentrating and drying the second dialyzate 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 dangshen polysaccharide prepared according to the method described in the embodiment 1 and the second licorice polysaccharide prepared according to the method described in the embodiment 2 are uniformly mixed in a weight ratio of 3:1 to obtain the active polysaccharide composition.
Comparative example 1 preparation of active polysaccharide composition
The third dangshen polysaccharide prepared according to the method described in the embodiment 1 and the licorice crude polysaccharide prepared according to the method described in the embodiment 2 are uniformly mixed in a weight ratio of 3:1 to obtain the active polysaccharide composition.
Comparative example 2 preparation of active polysaccharide composition
The third dangshen polysaccharide prepared according to the method described in the embodiment 1 and the first licorice polysaccharide prepared according to the method described in the embodiment 2 are uniformly mixed in a weight ratio of 3:1 to obtain the active polysaccharide composition.
Comparative example 3 preparation of active polysaccharide composition
The third dangshen polysaccharide prepared according to the method of the embodiment 1 and the third licorice polysaccharide prepared according to the method of the embodiment 2 are uniformly mixed in a weight ratio of 3:1 to obtain the active polysaccharide composition.
Experimental example
88 balb/c mice weighing 20+ -2 g were taken, 44 male and female mice each, and the male and female mice were randomly divided into 11 groups of 8 mice each, which were respectively a blank group, a model group, and an experimental group. The blank and model groups were given equal volumes of 0.75% nacl solution; the model was separately administered with 0.75% NaCl solution containing the active polysaccharide or the active polysaccharide composition at a dose of 5mg/kg; the administration was performed by intragastric administration once daily for 7 consecutive days.
After 1h of last dose, each group of mice was given 300 μl of 50% ethanol by gavage. After ethanol injury for 2h, mice were sacrificed, dissected, taken out of the whole stomach, washed with normal saline, immersed in a 2% formalin solution for fixation for 10min, and then the gastric ulcer area was determined 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 the active polysaccharide composition administered in the experimental group is respectively: the third dangshen polysaccharide (i.e., dangshen polysaccharide according to the present invention) prepared according to the method of example 1, dangshen crude polysaccharide, first dangshen polysaccharide, second dangshen polysaccharide, and fourth dangshen polysaccharide, 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 polysaccharide or active polysaccharide composition on acute gastric injury caused by ethanol
In comparison with the blank set of the cells, ## p < 0.01, compared with the model group, ** p＜0.01， * p＜0.05。
as can be seen from Table 1, after the mice were perfused with gastric alcohol, the gastric ulcer index of the mice was significantly increased, indicating that acute drinking had a great damage to the mice' stomach.
As can be seen from the data of table 1, the third dangshen polysaccharide (i.e., dangshen polysaccharide according to the present invention), the dangshen crude polysaccharide, the first dangshen polysaccharide, the second dangshen polysaccharide, and the fourth dangshen polysaccharide, which were prepared according to the method of example 1, were greatly different in gastric ulcer index from mice; the index of gastric ulcer of mice of the third dangshen polysaccharide (i.e. dangshen polysaccharide) prepared according to the method of the embodiment 1 is obviously lower than that of dangshen crude polysaccharide, and is also obviously lower than that of the first dangshen polysaccharide, the second dangshen polysaccharide and the fourth dangshen polysaccharide. Furthermore, as can be seen from the data in table 1, the index of the first Codonopsis pilosula polysaccharide to the gastric ulcer of mice is higher than that of the Codonopsis pilosula polysaccharide. This illustrates: 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 effect of preventing or treating gastric injury; compared with the crude polysaccharide of codonopsis pilosula, the effect of preventing or treating gastric injury can be obviously improved; compared with the crude polysaccharide of radix codonopsis, the other radix codonopsis polysaccharide obtained in the preparation process of the invention can not obviously improve the effect of preventing or treating gastric injury, and even reduce the effect.
As can be seen from the data in table 1, the active polysaccharide composition prepared in example 3 has a much smaller gastric ulcer index on mice than the third dangshen polysaccharide prepared according to the method in example 1 (i.e., dangshen polysaccharide of the present invention), and is also much smaller than dangshen crude polysaccharide; however, the gastric ulcer index of the active polysaccharide compositions prepared in comparative examples 1 to 3 was not reduced as compared with the third dangshen polysaccharide prepared according to the method of example 1 (i.e., dangshen polysaccharide according to the present invention). This illustrates: only the active polysaccharide composition obtained by combining the third dangshen polysaccharide prepared by the method of the invention and the second licorice polysaccharide prepared by the method of the invention has the effect of preventing or treating gastric injury, which is further and greatly improved compared with the third dangshen polysaccharide. However, the active polysaccharide composition obtained by combining the third dangshen polysaccharide prepared by the above method according to the present invention with other licorice polysaccharide prepared by the method according to the present invention does not further improve the stomach injury preventing or treating effect of the third dangshen polysaccharide.
Claims (7)
1. An active polysaccharide composition, which is characterized in that the active polysaccharide composition comprises codonopsis pilosula polysaccharide and licorice polysaccharide; wherein the weight ratio of the codonopsis pilosula polysaccharide to the liquorice polysaccharide is 3:1;
the codonopsis pilosula polysaccharide is prepared by a method comprising the following steps:
s11, taking traditional Chinese medicine codonopsis pilosula, and then preparing codonopsis pilosula crude polysaccharide by using a water extraction and alcohol precipitation method;
s12, dissolving the codonopsis pilosula crude polysaccharide in water, and then dialyzing by using a dialysis bag with the aperture of 18-22 kD to obtain
Concentrating and drying the first trapped fluid to obtain first dangshen polysaccharide;
s13, dialyzing the first dialysate by using a dialysis bag with the size of 8-12 kD to obtain second dialysate and second trapped fluid, concentrating and drying the second trapped fluid to obtain second codonopsis pilosula polysaccharide;
s14, dialyzing the second dialyzate by using a dialysis bag with a molecular weight of 2-4 kD to obtain a third dialyzate and a third trapped fluid, concentrating and drying the third trapped fluid to obtain third dangshen polysaccharide; concentrating and drying the third dialysate to obtain fourth codonopsis pilosula polysaccharide; taking the third dangshen polysaccharide to obtain the dangshen polysaccharide;
the liquorice polysaccharide is prepared by a method comprising the following steps:
s21, preparing licorice, namely extracting and precipitating the licorice by water to prepare crude polysaccharide of the licorice;
s22, dissolving the licorice crude polysaccharide in water, and then dialyzing with a dialysis bag with the aperture of 14-16 kD to obtain
Concentrating and drying the first trapped fluid to obtain first glycyrrhiza polysaccharide;
s23, dialyzing the first dialysate by using a 7-9 kD dialysis bag to obtain a second dialysate and a second trapped fluid, concentrating and drying the second trapped fluid to obtain second glycyrrhiza polysaccharide; concentrating and drying the second dialyzate to obtain third glycyrrhiza polysaccharide; and taking the second glycyrrhiza polysaccharide to obtain the glycyrrhiza polysaccharide.
2. The active polysaccharide composition according to claim 1, wherein the dialysis is performed in step S12 with a dialysis bag having a pore size of 20 kD.
3. The active polysaccharide composition according to claim 1, wherein S13 is dialyzed with a dialysis bag having a pore size of 10 kD.
4. The active polysaccharide composition according to claim 1, wherein the dialysis is performed in step S14 with a dialysis bag having a pore size of 3 kD.
5. The active polysaccharide composition according to claim 1, wherein in step S22 the dialysis is performed with a dialysis bag having a pore size of 15 kD.
6. The active polysaccharide composition according to claim 1, wherein the dialysis is performed in step S23 with a dialysis bag having a pore size of 8 kD.
7. Use of the active polysaccharide composition according to any one of claims 1 to 6 for the preparation of a product having a gastric injury preventing or treating effect;
the gastric injury is acute alcoholic gastric injury.
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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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- 2022-07-28 CN CN202210899455.6A patent/CN115286721B/en active Active
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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