CN115737752A - Application of rhizoma bletillae aqueous extract - Google Patents
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Abstract
The invention discloses an application of rhizoma bletillae water extract in preparing a medicament or health food for treating and/or preventing alcoholic liver injury. The rhizoma bletillae water extract has a good effect in treating and/or preventing alcoholic liver injury.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine application, and particularly relates to a new application of a rhizoma bletillae aqueous extract.
Background
Alcoholic liver injury (ALD) is one of common diseases seriously harming public health worldwide at present, the incidence rate rises year by year, and the ALD poses serious threats to the health of people and social development. Alcoholic liver injury is a liver cell structural abnormality and a dysfunction disease caused by long-term or large-scale drinking, the development of the alcoholic liver injury can be divided into four stages of alcoholic fatty liver, alcoholic hepatitis, hepatic fibrosis and liver cirrhosis, and even more, the alcoholic liver injury can cause liver cancer and cause death and other poor health outcomes.
At present, the treatment of alcoholic liver injury adopts the modes of alcohol withdrawal, nutrition support, little medicine intervention and the like, but specific medicines are still lacked. CN110101798A discloses an anti-alcohol and liver-protecting agent, which is prepared from kudzuvine flower, hovenia dulcis thunb, camphor tree seed, penthorum chinense pursh, bletilla striata, clematis armandii, radix saururi chinensis and liquidambar formosana through the steps of distillation, water extraction and the like. The anti-hangover and hepatoprotective agent has complicated medicinal components and large toxic and side effects.
In recent years, the medicine-food homologous traditional Chinese medicines are paid much attention because of the advantages of multiple physiological functions, low toxic and side effects, multiple ways and multiple target points, and part of effective parts are developed into food or medicines for treating and preventing related diseases, so the medicine-food homologous traditional Chinese medicines also become one of important medicines for preventing and treating alcoholic liver diseases and food research and development directions.
Bletilla striata is a dry tuber of Bletilla striata (thunb.) Reichb.f. of Orchidaceae, has the functions of astringency and hemostasis, detumescence and tissue regeneration, is often used for treating hemoptysis, hematemesis, traumatic hemorrhage and the like, and is an important medicine-food homologous traditional Chinese medicine. In recent years, the application of bletilla striata polysaccharide in treating acute alcoholic liver injury is reported, but the curative effect of the bletilla striata polysaccharide is still to be enhanced.
Disclosure of Invention
In view of the above, the invention provides an application of a water extract of rhizoma bletillae in preparing a medicament or health food for treating and/or preventing alcoholic liver injury, and the water extract has a good curative effect. Furthermore, the aqueous extract of bletilla striata has low toxicity. The bletilla polysaccharide is part of water-soluble components of bletilla, is usually prepared by water extraction and alcohol precipitation, and is purified to obtain the bletilla polysaccharide. Therefore, the bletilla polysaccharide is a chemical component which is soluble in water but not soluble in ethanol solution with certain concentration in the bletilla medicinal materials. At present, reports about the treatment of acute alcoholic liver injury by bletilla striata polysaccharides exist, but no evidence indicates that other chemical components in bletilla striata water-soluble components, especially alcohol-soluble components, also have an improvement effect on alcoholic liver injury. The research of the invention finds that the rhizoma bletillae aqueous extract has obviously better curative effect on the treatment of alcoholic liver injury compared with rhizoma bletillae polysaccharide, and can achieve better treatment effect under the same dosage.
Therefore, the invention provides an application of rhizoma bletillae aqueous extract in preparing a medicament or health food for treating and/or preventing alcoholic liver injury.
According to the use of the present invention, preferably, the alcoholic liver injury is chronic alcoholic liver injury.
Preferably, according to the use of the invention, the alcoholic liver injury is manifested as an increase in the level of alanine aminotransferase and/or aspartate aminotransferase in the serum.
According to the use of the present invention, preferably, the aqueous bletilla striata extract is used as the only active ingredient.
According to the use of the present invention, preferably, the aqueous extract of rhizoma bletillae is obtained by the following method:
(1) Extracting rhizoma bletillae with water as a solvent to obtain an aqueous extract;
(2) Concentrating the water extract under reduced pressure to obtain concentrated water extract;
(3) And drying the concentrated water extract to obtain a bletilla striata water extract.
According to the use of the present invention, preferably, the extraction manner is heating reflux extraction; the extraction times are 1 to 5; the dosage of the water extracted each time is 5 to 30 times of the weight of the bletilla striata medicinal materials; the extraction time is 0.5-5 h each time.
According to the use of the present invention, it is preferred that the aqueous extract solution is concentrated under reduced pressure at 40 to 70 ℃ to obtain a concentrated aqueous extract solution having a volume of 0.1 to 0.4 times the volume of the aqueous extract solution.
According to the application of the invention, preferably, the total polysaccharide content in the aqueous extract of rhizoma bletillae is more than or equal to 70.00wt%, the total protein content is more than or equal to 4.00wt%, and the alcohol-soluble extract content is more than or equal to 10.00wt%.
According to the use of the invention, preferably, the human dosage of the rhizoma bletillae aqueous extract is 2-20 mg/kg.
According to the use of the invention, preferably, the human dosage of the rhizoma bletillae aqueous extract is 4-12 mg/kg.
The common bletilla pseudobulb as a medicine-food homologous traditional Chinese medicine has small toxic and side effects. Although the bletilla striata polysaccharide is considered as an important bioactive component in the bletilla striata all the time, the invention unexpectedly discovers that the dosage of the aqueous bletilla striata extract is lower than that of the bletilla striata polysaccharide, and the curative effect is better, which shows that other water-soluble components in the aqueous bletilla striata extract can be used alone or in synergistic effect with the bletilla striata polysaccharide to have the effect of treating alcoholic liver injury.
Drawings
FIG. 1 is a bar graph of the mean ALT enzyme activity in serum of mice in each group after administration.
FIG. 2 is a bar graph showing the average value of AST enzyme activities in the serum of each group of mice after administration.
FIG. 3 is a H & E staining chart of liver tissue sections of various groups of mice after administration.
In fig. 1 and 2, P <0.05 and P <0.01 are shown.
Detailed Description
The present invention will be further described with reference to the following specific examples, but the scope of the present invention is not limited thereto.
The invention unexpectedly discovers that the aqueous extract of bletilla has good curative effect on the aspect of treating alcoholic liver injury, and the effect is superior to that of bletilla polysaccharide. Therefore, the invention provides an application of rhizoma bletillae aqueous extract in preparing a medicament or health food for treating and/or preventing alcoholic liver injury. The prior literature discloses that rhizoma bletillae polysaccharide can be used for treating acute alcoholic liver injury, but the literature does not disclose the application of rhizoma bletillae water extract in treating alcoholic liver injury. The bletilla striata polysaccharide is considered as a main active ingredient in the bletilla striata medicinal material, and the invention unexpectedly discovers that the aqueous bletilla striata extract has better curative effect on alcoholic liver injury compared with the bletilla striata polysaccharide. This shows that other chemical components in the aqueous extract of bletilla striata can act alone or in synergy with bletilla striata polysaccharide to have the effect of treating alcoholic liver injury.
The alcoholic liver injury is preferably chronic alcoholic liver injury. In certain embodiments, the alcoholic liver injury is caused by an increase in serum alanine aminotransferase content and/or aspartate aminotransferase content. Aspartate aminotransferase, also known as "AST," is found primarily in organs such as the liver, heart muscle, kidney, etc. Alanine aminotransferase, also known as "ALT", is found primarily in the liver. ALT is more sensitive than AST changes, ranging from mild steatosis in the liver to fibrosis or even cancer, and changes with disease progression.
Preferably, the rhizoma bletillae aqueous extract is used as the only active ingredient. Thus, not only can a good curative effect be obtained, but also the toxic and side effects can be reduced.
The content of total polysaccharide in the aqueous extract of rhizoma bletillae is more than or equal to 70.00wt%; preferably 70 to 80wt%; more preferably 73 to 77wt%. Therefore, the curative effect of the bletilla striata water extract on alcoholic liver injury can be improved.
The total protein content in the aqueous extract of rhizoma bletillae is more than or equal to 4.00wt%; preferably 4 to 6wt%; more preferably 4.5 to 5wt%. Therefore, the curative effect of the bletilla striata water extract on alcoholic liver injury can be improved.
The content of alcohol-soluble extract in the aqueous extract of rhizoma bletillae is more than or equal to 10.00wt%; preferably 10 to 13wt%; more preferably 11 to 12wt%. Therefore, the curative effect of the bletilla striata water extract on alcoholic liver injury can be improved.
The bletilla striata water extract can be obtained by adopting the following method: (1) Extracting rhizoma bletillae with water as a solvent to obtain an aqueous extract; (2) concentrating the water extract under reduced pressure to obtain a concentrated water extract; and (3) drying the concentrated water extract to obtain a rhizoma bletillae water extract.
In the present invention, the extraction method may be heating reflux extraction. The extraction times can be 1-5 times; preferably 1 to 3 times; more preferably 2 to 3 times.
The amount of the water used for each extraction is 5 to 30 times, preferably 10 to 30 times, more preferably 12 to 24 times, and still more preferably 14 to 20 times of the mass of the rhizoma bletillae. Taking 500g of bletilla striata medicinal materials as a reference, and the using amount of water extracted each time is 2.5-15L; preferably, the amount of water used per extraction is 5 to 15L, more preferably 6 to 12L, still more preferably 7 to 10L.
The extraction time can be 0.5-5 h each time; preferably 0.5 to 2 hours; more preferably 0.5 to 1 hour.
The temperature of the reduced pressure concentration can be 40-70 ℃; preferably 50 to 60 ℃; more preferably 50 to 55 deg.c.
The volume of the concentrated water extract can be 0.1-0.4 times of the volume of the water extract; preferably, the volume of the concentrated aqueous extract is 0.1 to 0.3 times the volume of the aqueous extract; more preferably, the volume of the concentrated aqueous extract solution is 0.2 to 0.3 times the volume of the aqueous extract solution.
The drying temperature can be 40-80 ℃; preferably 50 to 70 ℃; more preferably 60 to 70 ℃.
In the invention, the human dosage of the rhizoma bletillae water extract is 2-20 mg/kg; preferably 4 to 12mg/kg; more preferably 6 to 10mg/kg. Thus, a good therapeutic effect can be obtained with a low dose, and toxic side effects can be reduced.
Example 1
Heating and refluxing 500g of dried rhizoma bletilla sheet as a solvent by using 10L of water as a solvent for 2 hours, extracting for 2 times, filtering, and combining filtrates to obtain a water extract.
Concentrating the water extractive solution at 50 deg.C under reduced pressure to obtain concentrated water extractive solution; the volume of the concentrated aqueous extract was 0.2 times the volume of the aqueous extract.
Drying the concentrated water extract at 60 deg.C to obtain rhizoma bletilla water extract.
Example 2
Example 1 was repeated except that the amount of extract water used was 7.5L per heating reflux.
Example 3
The same procedure as in example 1 was repeated, except that the number of extractions was 3.
Example 4
The same procedure as in example 1 was repeated, except that the extraction time was 3 hours.
Preparation example-preparation of bletilla polysaccharide
Taking the bletilla striata water extract obtained in example 1, adding water into the bletilla striata water extract, then adding 95vol% ethanol until the ethanol concentration is 80vol%, standing overnight at 4 ℃, then centrifuging for 30min at the rotating speed of 4000r/min, and discarding the supernatant to obtain a precipitate. And (4) freeze-drying the precipitate to obtain the bletilla striata polysaccharide.
Experimental example 1
Calculating the extract yield of the bletilla striata water extracts obtained in the embodiments 1-4, and determining the total polysaccharide content, the total protein content and the alcohol-soluble extract content in the bletilla striata water extracts, wherein the specific method is as follows:
the extract yield of the rhizoma bletillae water extract is as follows: the measurement was carried out by the hot dipping method in "2201 extract measurement method" in the fourth part of the pharmacopoeia of China (2020 edition).
The total polysaccharide content in the rhizoma bletillae water extract is as follows: the reference substance is anhydrous glucose measured by a phenol-sulfuric acid method.
Total protein content in aqueous extract of bletilla striata: the control substance is bovine serum albumin determined by Coomassie brilliant blue method.
Alcohol-soluble extract content in rhizoma bletillae water extract: the measurement was carried out by an alcohol-soluble extract measurement method under the item "2201 extract measurement method" in the fourth part of the year 2020 edition "Chinese pharmacopoeia", using 95vol% ethanol as a solvent.
The results obtained are shown in table 1.
TABLE 1
Experimental example 2
1. Animals and groups
48 male C57BL/6J mice (7-8 weeks old, 20. + -.2 g weight, purchased from Beijing Wintolite laboratory animal technology Co., ltd., certification number: SCXK (Jing) 2016-0011) were bred in an SPF-level environment (23. + -. 2 ℃ C., relative humidity 55. + -. 5%), observed daily to record the general condition, and weighed every three days. After one week of adaptive feeding, the animals were randomly divided into a normal group (Con), a model group (Mod), a positive drug silymarin group (Pos), a low-dose group (BSPL) of bletilla polysaccharide, a medium-dose group (BSPM) of bletilla polysaccharide, a high-dose group (BSPH) of bletilla polysaccharide, a low-dose group (BSWL) of bletilla water extract and a high-dose group (BSWH) of bletilla water extract, for a total of 8 groups, and 6 animals were each group.
2. Modeling and administration
A Lieber-Decalli liquid feed pairing feeding method is adopted to establish a chronic alcoholic liver injury model. Con groups were fed with control liquid feed (no alcohol) and the remaining groups were fed with alcohol liquid feed (4 vol% alcohol) for 6 weeks. Then, gavage 0.2mL/d of physiological saline into Con and Mod groups every day; pos group is intragastrically administered with 100mg/kg silymarin; respectively intragastric feeding BSPL, BSPM and BSPH groups with rhizoma bletilla polysaccharide water solution (prepared by dissolving rhizoma bletilla polysaccharide water solution in water) of 100mg/kg, 200mg/kg and 400mg/kg (calculated on rhizoma bletilla polysaccharide); BSWL and BSWH groups were individually gavaged with aqueous rhizoma bletilla water extracts (obtained by dissolving the aqueous rhizoma bletilla water extracts of example 1 in water), and the gavaged amounts were 50mg/kg and 200mg/kg (both based on the aqueous rhizoma bletilla extracts), respectively, and the rearing was continued for 42 days.
3. Sample collection and processing
After the raising period is finished, the mice are fasted for 12 hours without water, and the mice are killed by taking off the neck after eyeballs are picked and blood is taken. After the whole blood is kept stand for about 2 hours, the whole blood is centrifuged at 3500r/min for 15min at 4 ℃, and serum is separated. ALT kit (batch No. 20220105) purchased from Nanjing institute of bioengineering is adopted, and the ALT content in serum is determined according to the method of the kit instruction. An AST kit (batch number: 20220104) purchased from Nanjing institute of bioengineering is adopted, and the content of AST in serum is measured according to a method of a kit instruction. Liver tissue was divided, and half of the largest leaves were selected and fixed with paraformaldehyde for pathological analysis of H & E staining.
4. Data processing
Data processing was performed using SPSS software and results were expressed as mean. + -. Standard deviation (Means. + -. SD). Differences between sample data were compared using analysis of variance (ANOVA) and multiple comparisons were performed using the LSD (Least Significant Difference) test. P <0.05 indicates significant difference, and P <0.01 indicates very significant difference.
FIG. 1 is a bar graph of the mean ALT enzyme activity in serum of mice in each group after administration. Compared with the Con group, the ALT content of the Mod group is remarkably increased, which indicates that the model building of the mouse chronic alcoholic liver injury model is successful. Significant differences (P < 0.05) are found in comparison with the Mod group in each administration group, and the rhizoma bletillae polysaccharide administration group and the rhizoma bletillae aqueous extract administration group have certain dose dependence. The BSPH group and BSWH group had very significant differences compared to the model group (P < 0.01); wherein, the BSWH group has the best effect and the effect is better than that of the silymarin group (100 mg/kg). ALT levels in the BSWH group (200 mg/kg) were significantly lower than in the BSPH group (400 mg/kg), indicating that the aqueous extract of bletilla striata was used in smaller amounts and with superior efficacy compared to the polysaccharides of bletilla striata.
FIG. 2 is a bar graph showing the average value of AST enzyme activity in serum of mice in each group after administration. Compared with the Con group, the AST content of the Mod group is obviously increased, which indicates that the model building of the mouse chronic alcoholic liver injury model is successful. Significant differences (P < 0.05) are found in comparison with the Mod group in each administration group, and the rhizoma bletillae polysaccharide administration group shows certain dose dependence. Compared with the rhizoma bletillae polysaccharide group, the rhizoma bletillae aqueous extract group can obtain better curative effect under lower medicinal dosage.
FIG. 3 is a H & E staining chart of liver tissue sections of various groups of mice after administration. As seen from FIG. 3, in the Mod group, there were a large number of vacuoles in the cytoplasm, uneven cell arrangement, lipid droplets in the hepatocytes, a large number of lipid droplets, and severe fat accumulation accompanied by inflammatory cell infiltration, as compared with the Con group. The volume and the number of lipid droplets in the liver cells of each administration group are reduced, but the cells of the low dose group and the medium dose group still have different edema and degeneration, and the liver cord arrangement is also relatively disordered and is accompanied by a small amount of inflammatory cell infiltration. Meanwhile, the fatty degeneration of liver cells of the Pos group, the BSPH group and the BSWH group is improved more obviously, liver cords are arranged neatly, and the tissue form is close to that of the Con group.
The present invention is not limited to the above-described embodiments, and any variations, modifications, and substitutions which may occur to those skilled in the art may be made without departing from the spirit of the invention.
Claims (10)
1. An application of rhizoma bletilla water extract in preparing medicine or health food for treating and/or preventing alcoholic liver injury is provided.
2. The use of claim 1, wherein the alcoholic liver injury is chronic alcoholic liver injury.
3. Use according to claim 1, wherein the alcoholic liver injury is manifested as an increase in serum alanine aminotransferase and/or aspartate aminotransferase levels.
4. The use as claimed in claim 1, wherein the aqueous bletilla extract is the sole active ingredient.
5. The use according to claim 1, wherein the aqueous bletilla extract is obtained by the following method:
(1) Extracting rhizoma bletillae with water as a solvent to obtain an aqueous extract;
(2) Concentrating the water extract under reduced pressure to obtain concentrated water extract;
(3) And drying the concentrated water extract to obtain the rhizoma bletillae water extract.
6. Use according to claim 5, characterized in that the extraction mode is a heating reflux extraction; the extraction times are 1 to 5; the dosage of the water extracted each time is 5 to 30 times of the weight of the bletilla striata medicinal materials; the extraction time is 0.5-5 h each time.
7. The use as claimed in claim 5, wherein the aqueous extract solution is concentrated at 40 to 70 ℃ under reduced pressure to give a concentrated aqueous extract solution having a volume of 0.1 to 0.4 times the volume of the aqueous extract solution.
8. The use as claimed in claim 1, wherein the water extract of rhizoma bletilla contains polysaccharide 70.00wt%, total protein 4.00wt%, and alcohol-soluble extract 10.00wt%.
9. Use according to any one of claims 1 to 8, wherein the human dose of the aqueous extract of bletilla striata is between 2 and 20mg/kg.
10. The use as claimed in claim 9, wherein the human amount of the rhizoma bletillae aqueous extract is 4-12 mg/kg.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429254A (en) * | 2008-12-05 | 2009-05-13 | 云南植物药业有限公司 | Bletilla striata polysaccharide, preparation method and new uses thereof |
| CN104042656A (en) * | 2014-06-27 | 2014-09-17 | 漳州片仔癀药业股份有限公司 | Application of Pien Tze Huang and preparation thereof in preparing medicines for treating alcoholic liver injury |
| CN105919992A (en) * | 2016-05-03 | 2016-09-07 | 苏州大学 | Application of apigenin in preparing medicines or health care foods for preventing and/or treating alcoholic liver injury |
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- 2022-11-24 CN CN202211482321.0A patent/CN115737752A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101429254A (en) * | 2008-12-05 | 2009-05-13 | 云南植物药业有限公司 | Bletilla striata polysaccharide, preparation method and new uses thereof |
| CN104042656A (en) * | 2014-06-27 | 2014-09-17 | 漳州片仔癀药业股份有限公司 | Application of Pien Tze Huang and preparation thereof in preparing medicines for treating alcoholic liver injury |
| CN105919992A (en) * | 2016-05-03 | 2016-09-07 | 苏州大学 | Application of apigenin in preparing medicines or health care foods for preventing and/or treating alcoholic liver injury |
Non-Patent Citations (2)
| Title |
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| 孔伟华等: "白及化学成分、药理作用和白及多糖提取工艺的研究进展", 《中医药信息》, pages 1 - 3 * |
| 贺国芳等: "白及多糖对小鼠急性酒精性肝损伤的保护作用", 《中国医院药学杂志》, pages 1659 * |
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