Method for improving utilization rate and curative effect of active ingredients of rhizoma polygonati medicinal material for preventing and treating liver injury

Abstract

The invention relates to the technical field of medicinal material treatment, and discloses a method for improving the utilization rate and curative effect of active ingredients of rhizoma polygonati medicinal materials for preventing and treating liver injury, the method comprises the following steps: extracting and concentrating rhizoma polygonati medicinal materials with water, adding 4 times of volume of absolute ethyl alcohol to enable the final concentration of the ethyl alcohol to reach 80%, separating out polysaccharide, and performing centrifugal separation; freeze-drying the polysaccharide precipitate to obtain a polygonatum polysaccharide component; concentrating, freezing and drying the supernate to obtain a non-polysaccharide component enriched with low-content effective components such as flavone, saponin and alkaloid; finally, the polysaccharide component and the non-polysaccharide component are mixed according to the mass ratio of 10: 90 or 30: 70 for use. The anti-liver injury effect of the rhizoma polygonati water extract is superior to that of the traditional rhizoma polygonati water extract, and the liver index and the serum transaminase level during liver injury can be remarkably reduced. According to the technical scheme, the method is easy to implement and low in cost, and the utilization rate of low-content pharmacological active substances in rhizoma polygonati is remarkably increased.

Description

Technical Field

[0001] This invention relates to the field of medicinal material processing technology, and more specifically, to a method for improving the utilization rate and efficacy of active ingredients in Polygonatum sibiricum for the prevention and treatment of liver damage. Background Technology

[0002] With the development of intensive animal husbandry and the impact of environmental factors, liver injury in humans and livestock caused by drug poisoning, moldy feed, and pathogenic microorganism infections is becoming increasingly common. Liver injury can cause a series of pathological changes, including metabolic disorders, bile formation and excretion disorders, and reduced detoxification capacity, posing serious threats to human health and the livestock industry. Therefore, the development of low-toxicity and low-residue drugs for the effective protection and treatment of liver injury is of significant practical importance. With the modernization of traditional Chinese medicine and the continuous development of its pharmacology, the use of traditional Chinese medicine to prevent and treat liver injury is receiving increasing attention.

[0003] Polygonatum, a traditional and widely used medicinal herb in my country, possesses pharmacological effects such as enhancing immunity, lowering blood sugar, anti-tumor activity, antioxidant properties, and liver protection. Studies have shown that water extracts and polysaccharides of Polygonatum have a certain protective effect against liver damage. The effective components of Polygonatum include polysaccharides, steroidal saponins, alkaloids, flavonoids, and phenylpropanoids, among other bioactive substances. Polysaccharides are the most abundant, accounting for 10%-20% of the dry weight of the product. Although flavonoids, saponins, and alkaloids in Polygonatum have significant pharmacological effects, their low content and high extraction costs have hindered their industrial-scale development and utilization. In traditional methods of utilizing Polygonatum, the pharmacological effects of low-content pharmacologically active substances such as flavonoids, saponins, and alkaloids may not be fully realized, resulting in a waste of valuable resources. Currently, how to further develop and utilize the various pharmacologically active substances in Polygonatum, especially those with low content, efficiently and at low cost remains a pressing technical problem. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides a method for improving the utilization rate and efficacy of active ingredients in Polygonatum sibiricum, a medicinal herb for preventing and treating liver damage, comprising the following steps: Step 1: Grind and sieve the Polygonatum medicinal material, extract it with water at 80℃ for 1 hour at a material-to-liquid ratio of 1:20, repeat the extraction 3 times, combine the filtrates and concentrate under reduced pressure. Step 2: Add 4 times the volume of anhydrous ethanol to the concentrated Polygonatum water extract to make the final ethanol concentration in the solution 80%. Let it stand overnight at 4°C to precipitate polysaccharides. Centrifuge to separate the polysaccharide precipitate and supernatant. Step 3: Dissolve the polysaccharide precipitate in water and freeze-dry it to obtain the Polygonatum polysaccharide component; Step 4: After concentrating the supernatant under reduced pressure, freeze-dry it to obtain a non-polysaccharide component of Polygonatum odoratum enriched with low-content effective components such as flavonoids, saponins, and alkaloids. Step 5: Mix the Polygonatum polysaccharide component with the non-polysaccharide component at a mass ratio of 10:90 or 30:70.

[0005] Preferably, the Polygonatum medicinal material is Polygonatum multiflorum, which is dried at 65°C for 5 hours and then pulverized through a 60-mesh sieve.

[0006] Preferably, the water extraction process uses a constant temperature magnetic stirrer for extraction, and after extraction, the filtrate is collected by filtering with four layers of gauze, and then the filtrate is filtered under reduced pressure using a Buchner funnel.

[0007] Preferably, the vacuum concentration is carried out at 50°C.

[0008] Preferably, the centrifugation conditions during the alcohol precipitation process are 5000 rpm for 10 minutes.

[0009] Preferably, the sample is frozen into a solid state at -20°C before freeze-drying.

[0010] Preferably, spray drying is used instead of freeze drying in industrial production.

[0011] Preferably, the dosage of the mixture is 300 mg / kg body weight.

[0012] Preferably, the method is used to prepare pharmaceutical preparations, health products, or feed additives for the prevention and treatment of liver damage.

[0013] Preferably, the protective effects of different ratios are evaluated using a carbon tetrachloride-induced acute liver injury model in mice, and the optimal ratio is selected.

[0014] The beneficial effects of this invention are as follows: Significantly improved efficacy in preventing and treating liver injury: Validation using a carbon tetrachloride (CCL4)-induced acute liver injury model in mice showed that when the polysaccharide and non-polysaccharide components of Polygonatum were mixed at a ratio of 10:90 or 30:70, the liver index showed a significant decreasing trend compared to the traditional Polygonatum aqueous extract group (p<0.1), indicating a superior anti-liver injury effect. The liver index in all treatment groups was significantly lower than that in the model group (p<0.01), and serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were significantly reduced, demonstrating better hepatoprotective effects.

[0015] Significantly improves resource utilization: This invention enriches the low-content non-polysaccharide active ingredients (flavonoids, saponins, alkaloids, etc.) in the alcohol precipitation supernatant, so that the active substances that were originally difficult to exert their medicinal effects due to low content can be fully utilized, solving the problem of waste of valuable resources in the traditional use of Polygonatum and realizing the in-depth development of Polygonatum medicinal material.

[0016] Reduced production costs: When using a 10:90 ratio, the amount of Polygonatum polysaccharides used is only 10% of that used in traditional methods, significantly reducing the amount of polysaccharide components and production costs. Furthermore, the process is simple and easy to implement, and spray drying technology can be used to replace freeze drying in industrial production, further saving production costs.

[0017] Achieving scientific formulation: The scientific formulation selected based on the results of drug efficacy experiments not only achieves better synergistic effects in drug efficacy, but also overcomes the defect that the ratio of polysaccharide and non-polysaccharide components in natural "compound" may not be optimal, and gives full play to the synergistic effect between drugs.

[0018] With broad application prospects, this invention can be used in the traditional Chinese medicine preparation, health product and feed additive industries. It provides new technical approaches and ideas for improving the utilization rate of Polygonatum and other traditional Chinese medicine resources (especially low-content pharmacologically active substances), and has important industrial promotion value. Detailed Implementation

[0019] The subject matter described herein will now be discussed with reference to exemplary embodiments. It should be understood that these embodiments are discussed only to enable those skilled in the art to better understand and implement the subject matter described herein, and changes may be made to the function and arrangement of the elements discussed without departing from the scope of this specification. Various processes or components may be omitted, substituted, or added as needed in the examples. Furthermore, some features described in the examples may be combined in other examples.

[0020] Example 1 This embodiment proposes a method to improve the utilization rate and efficacy of the active ingredients in Polygonatum sibiricum for preventing and treating liver damage, including the following steps: Step 1: Grind and sieve the Polygonatum medicinal material, extract it with water at 80℃ for 1 hour at a material-to-liquid ratio of 1:20, repeat the extraction 3 times, combine the filtrates and concentrate under reduced pressure. Step 2: Add 4 times the volume of anhydrous ethanol to the concentrated Polygonatum water extract to make the final ethanol concentration in the solution 80%. Let it stand overnight at 4°C to precipitate polysaccharides. Centrifuge to separate the polysaccharide precipitate and supernatant. Step 3: Dissolve the polysaccharide precipitate in water and freeze-dry it to obtain the Polygonatum polysaccharide component; Step 4: After concentrating the supernatant under reduced pressure, freeze-dry it to obtain a non-polysaccharide component of Polygonatum odoratum enriched with low-content effective components such as flavonoids, saponins, and alkaloids. Step 5: Mix the Polygonatum polysaccharide component and the non-polysaccharide component at a mass ratio of 10:90.

[0021] in: The medicinal material is Polygonatum multiflorum, which is dried at 65℃ for 5 hours and then pulverized through a 60-mesh sieve.

[0022] The water extraction process uses a constant temperature magnetic stirrer. After extraction, the filtrate is collected by filtering with four layers of gauze and then filtered under reduced pressure using a Buchner funnel.

[0023] Concentration under reduced pressure was carried out at 50°C.

[0024] The centrifugation conditions during the alcohol precipitation process were 5000 rpm for 10 minutes.

[0025] Before freeze-drying, the sample was frozen to a solid state at -20°C.

[0026] In industrial production, spray drying is used to replace freeze drying.

[0027] The dosage of the mixture is 300 mg / kg body weight.

[0028] The method is used to prepare pharmaceutical preparations, health products, or feed additives for the prevention and treatment of liver injury.

[0029] Example 2 The difference between this embodiment and Embodiment 1 is that the Polygonatum polysaccharide component and the non-polysaccharide component are mixed at a mass ratio of 30:70.

[0030] Example 3 Preparation of Polysaccharide and Non-Polysaccharide Components of Polygonatum: The dried (65℃, 5 hours) Polygonatum odoratum herb was pulverized and passed through a 60-mesh sieve. An appropriate amount of herb was weighed and extracted at a 1:20 material-to-liquid ratio in a magnetic stirrer at 80℃ for 1 hour. The extract was filtered through four layers of gauze and the filtrate was collected. The filtrate was then filtered again under reduced pressure using a Buchner funnel and the filtrate was collected. The extraction process was repeated twice on the residue. The filtrates from the three extractions were combined and concentrated under reduced pressure at 50℃ to a certain volume. The concentrate was dispensed into petri dishes and frozen at -20℃ to solidify. The solid extract was then freeze-dried in a freeze dryer to obtain the Polygonatum odoratum water extract. The concentrated Polygonatum odoratum water extract was then mixed with 4 times its volume of anhydrous ethanol to achieve a final ethanol concentration of 80%. The solution was allowed to stand overnight at 4℃ to precipitate polysaccharides. The precipitate was centrifuged at 5000 rpm for 10 minutes and collected as crude Polygonatum odoratum polysaccharide. This precipitate was dissolved in an appropriate amount of water, frozen at -20℃ to solidify, and then freeze-dried in a vacuum freeze dryer to obtain the Polygonatum odoratum polysaccharide fraction. The supernatant obtained after alcohol precipitation during the extraction of Polygonatum polysaccharides was concentrated to a certain volume under reduced pressure at 50°C and then freeze-dried to obtain the non-polysaccharide component of Polygonatum.

[0031] Screening and verification of the optimal ratio: One hundred and twelve male Kunming mice were randomly divided into four groups: a normal control group, a model group, a positive control group (biphenyl diester), a Polygonatum sibiricum water extract group, and groups with different ratios of Polygonatum sibiricum polysaccharide and non-polysaccharide components (polysaccharide component: non-polysaccharide component = 100:0, 90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90, 0:100), with eight mice per group. The oral gavage dose of Polygonatum sibiricum aqueous extract was 600 mg / kg BW. Mice in groups with different ratios of Polygonatum sibiricum polysaccharide and non-polysaccharide components were administered the corresponding mixture of polysaccharide and non-polysaccharide components daily by gavage at a dose of 300 mg / kg BW. The positive control group was administered biphenyl diester daily by gavage at a dose of 200 mg / kg BW. The normal control group and the model group were administered an equal volume of distilled water (10 mL / kg BW) daily by gavage. Administration was repeated once daily for 14 days. Two hours after the last administration, the normal control group was intraperitoneally injected with peanut oil (10 mL / kg BW), while the other groups were intraperitoneally injected with a 0.2% CCL4 solution prepared with peanut oil at a dose of 10 mL / kg BW.

[0032] Sixteen hours after intraperitoneal injection, mice in each group were weighed (fasted for 12 hours, but allowed water). Then, about 0.6 mL of blood was collected from the orbital cavity, allowed to stand at room temperature for 2 hours, and centrifuged at 2000 g for 15 min at 2–8 ℃. The supernatant was collected, and the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected using a fully automated biochemical analyzer.

[0033] After blood collection, the patient was euthanized by cervical dislocation. The liver was removed, rinsed with saline, dried, weighed, and its liver index was calculated. Liver index = liver weight / body weight × 100%.

[0034] Data for each group are expressed as mean ± standard deviation, and one-way ANOVA and Duncan's multiple comparisons were performed. Experimental results: The liver index of each group of mice is shown in Table 1. The liver index can directly reflect the degree of liver swelling during liver inflammation. The larger the liver index, the more severe the liver swelling, that is, the stronger the liver inflammatory response. As shown in Table 1, the liver index of the CCL4-induced acute hepatitis model group was significantly higher than that of the normal group (p<0.01), indicating that the acute hepatitis model was successfully established. The liver index of each treatment group was significantly lower than that of the model group (p<0.01), indicating that each treatment group achieved a good effect in alleviating acute hepatitis. There was no significant difference in the liver index between the different polysaccharide components and non-polysaccharide components of Polygonatum sibiricum compared with the water extract group of Polygonatum sibiricum (p>0.05), but the 30:70, 10:90 and 100:0 groups showed a trend of decreasing the liver index (p<0.1), indicating that the polysaccharide components:non-polysaccharide components of Polygonatum sibiricum at ratios of 30:70, 10:90 and 100:0 were more effective in alleviating acute liver injury than the water extract of Polygonatum sibiricum.

[0035] Table 1 Comparison of liver index among different groups of mice (x±SD, n=8) The serum ALT and AST activities of mice in each group are shown in Tables 2 and 3, respectively. ALT is mainly found in hepatocytes. When hepatocytes are damaged (e.g., due to inflammation or necrosis), a large amount is released into the blood, leading to an increase in the activity of this enzyme in serum. Therefore, it is one of the important indicators for clinical assessment of liver damage. AST is mainly found in hepatocytes, cardiomyocytes, and muscle cells, and is often used together with ALT to assess liver damage. As can be seen from Tables 2 and 3, the changes in serum ALT and AST activities in each group of mice were basically consistent with the changes in liver index.

[0036] Table 2 Comparison of serum ALT activity in mice of different groups (x-±SD, n=8) Table 3 Comparison of serum AST activity in mice of different groups (x-±SD, n=8) The results showed that the anti-liver damage effect was best when the polysaccharide component to non-polysaccharide component ratio of Polygonatum was 100:0, 30:70, and 10:90. Among these, the 10:90 and 30:70 ratios significantly reduced the amount of polysaccharide used while ensuring efficacy. This invention is simple, easy to implement, and low in cost. It can be used in the traditional Chinese medicine preparation, health product, and feed additive industries, providing a new technical approach to improve the utilization rate of low-content pharmacologically active substances in Polygonatum and other traditional Chinese medicine resources.

[0037] The embodiments of the present invention have been described above. However, the embodiments are not limited to the specific implementation methods described above. The specific implementation methods described above are merely illustrative and not restrictive. Those skilled in the art can make more equivalent embodiments under the guidance of the present embodiments, and all of them are within the protection scope of the present embodiments.

Claims

1. A method for improving the utilization rate and efficacy of active ingredients in Polygonatum sibiricum for preventing and treating liver damage, characterized in that, Includes the following steps: Step 1: Grind and sieve the Polygonatum medicinal material, extract it with water at 80℃ for 1 hour at a material-to-liquid ratio of 1:20, repeat the extraction 3 times, combine the filtrates and concentrate under reduced pressure. Step 2: Add 4 times the volume of anhydrous ethanol to the concentrated Polygonatum water extract to make the final ethanol concentration in the solution 80%. Let it stand overnight at 4°C to precipitate polysaccharides. Centrifuge to separate the polysaccharide precipitate and supernatant. Step 3: Dissolve the polysaccharide precipitate in water and freeze-dry it to obtain the Polygonatum polysaccharide component; Step 4: After concentrating the supernatant under reduced pressure, freeze-dry it to obtain a non-polysaccharide component of Polygonatum odoratum enriched with low-content effective components such as flavonoids, saponins, and alkaloids. Step 5: Mix the Polygonatum polysaccharide component with the non-polysaccharide component at a mass ratio of 10:90 or 30:

70.

2. The method according to claim 1, characterized in that, The medicinal material mentioned is Polygonatum multiflorum, which is dried at 65°C for 5 hours and then pulverized through a 60-mesh sieve.

3. The method according to claim 1, characterized in that, The water extraction process uses a constant temperature magnetic stirrer for extraction. After extraction, the filtrate is collected by filtering with four layers of gauze and then filtered under reduced pressure using a Buchner funnel.

4. The method according to claim 1, characterized in that, The vacuum concentration was carried out at 50°C.

5. The method according to claim 1, characterized in that, The centrifugation conditions during the alcohol precipitation process are 5000 rpm for 10 minutes.

6. The method according to claim 1, characterized in that, Before freeze-drying, the sample is frozen into a solid state at -20°C.

7. The method according to claim 1, characterized in that, In industrial production, spray drying is used to replace freeze drying.

8. The method according to claim 1, characterized in that, The dosage of the mixture is 300 mg / kg body weight.

9. The method according to claim 1, characterized in that, The method is used to prepare pharmaceutical preparations, health products, or feed additives for the prevention and treatment of liver damage.

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