Lipid-lowering traditional Chinese medicine composition, lipid-lowering traditional Chinese medicine extract and preparation method and application of lipid-lowering traditional Chinese medicine extract

Abstract

The invention provides a lipid-lowering traditional Chinese medicine composition and extract as well as a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicines. The lipid-lowering traditional Chinese medicine composition is prepared from the following raw materials: rhizoma cyperi and radix gentianae. The invention further provides an extract of the lipid-lowering traditional Chinese medicine composition. The extract is obtained by mixing rhizoma cyperi and radix gentianae and then co-extracting. The composition or the extract can be used for preparing medicines for preventing and / or treating hyperlipemia, and can be used for improving lipid metabolism disorder of liver cells and reducing the content of triglyceride and total cholesterol in the liver cells.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine compositions, specifically to a lipid-lowering traditional Chinese medicine composition, an extract thereof, a method for preparing the same, and its application in the prevention and / or treatment of hyperlipidemia. Background Technology

[0002] Hyperlipidemia is a common metabolic disease characterized by abnormally high levels of lipids such as cholesterol and triglycerides in the blood, and is a significant risk factor for cardiovascular and cerebrovascular diseases. Currently, the main drugs used clinically to treat hyperlipidemia are chemical drugs such as statins and fibrates. While these are effective, long-term use may lead to adverse reactions such as liver damage, muscle pain, and gastrointestinal discomfort, raising increasing concerns about their safety. Therefore, developing safe and effective lipid-lowering drugs is of significant clinical importance.

[0003] Traditional Chinese medicine (TCM) possesses unique advantages in lipid-lowering, and its multi-target and multi-pathway mechanisms of action provide new insights into the prevention and treatment of hyperlipidemia. Currently, numerous studies have explored the application of TCM in lipid-lowering. For example, CN117599142A discloses a TCM composition with preventive and therapeutic effects on non-alcoholic fatty liver disease (NAFLD). This composition includes Bupleurum chinense, Paeonia lactiflora, Curcuma longa, Glycyrrhiza uralensis, and Cyperus rotundus, which can significantly reverse disease symptoms in NAFLD mice. CN110101820B discloses a TCM composition for treating obesity, containing 8-12 parts of Cyperus rotundus. This composition not only reduces weight and lipids but also alleviates obesity-related complications such as fatty liver, hypertension, and hyperlipidemia.

[0004] Regarding liver protection, CN109568544A discloses a traditional Chinese medicine composition for treating acute and chronic hepatitis. This composition has strong antiviral efficacy and is beneficial in reducing the recurrence and deterioration rates of acute and chronic hepatitis. CN113499406A discloses a stomach-nourishing and weight-loss traditional Chinese medicine composition, which contains various traditional Chinese medicines such as Cyperus rotundus and has stomach-nourishing and weight-loss effects. In addition, CN113559210A discloses a traditional Chinese medicine composition with lipid-lowering effects, which contains various traditional Chinese medicines such as Gentiana scabra, Salvia miltiorrhiza, and Paeonia lactiflora.

[0005] Cyperus rotundus is a commonly used traditional Chinese medicine with the effects of regulating qi and relieving depression, regulating menstruation and relieving pain. Modern pharmacological studies have shown that it has anti-inflammatory and antioxidant effects. Gentiana scabra is bitter and cold in nature, with the effects of clearing heat and drying dampness, and purging liver and gallbladder fire. Clinically, it is often used to treat symptoms such as damp-heat in the liver and gallbladder. In existing technologies, the application of Cyperus rotundus and Gentiana scabra in lowering lipids is mostly as one ingredient in compound prescriptions, and there are no reports of Cyperus rotundus and Gentiana scabra being used alone in combination. Summary of the Invention

[0006] To address the problems of existing technologies, this invention provides a lipid-lowering traditional Chinese medicine composition, extract, preparation process, and application. The composition consists of Cyperus rotundus and Gentiana scabra, and the composition has a synergistic effect in lowering lipids. Furthermore, it was found that the combined extraction of the two herbs can significantly enhance their lipid-lowering activity and improve lipid metabolism disorders in hepatocytes.

[0007] To achieve the objectives of this invention, the following technical solution is adopted: In a first aspect, the present invention provides a lipid-lowering traditional Chinese medicine composition, which is composed of Cyperus rotundus and Gentiana scabra, wherein the weight ratio of Cyperus rotundus to Gentiana scabra is 2:1-4. Preferably, the lipid-lowering traditional Chinese medicine composition consists of Cyperus rotundus and Gentiana scabra in a weight ratio of 1:1.

[0008] Cyperus rotundus ( Cyperus rotundus L Cyperus rotundus is the dried rhizome of Cyperus rotundus, a plant belonging to the Cyperaceae family and the Cyperus genus. It is a commonly used traditional Chinese medicine with a long history of medicinal use and modern research value. Other names include Cyperus rotundus, ... etc.

[0009] Gentian is a plant of the Gentianaceae family (Gentiana scabra). Gentiana scabra Bunge ), Gentiana triflora ( G. triflora Pall. ), Gentiana linearifolia ( G. manshurica Kitag .) or Gentiana scabra ( G. rigescens Franch. The dried roots and rhizomes of ).

[0010] Secondly, the present invention provides an extract of the lipid-lowering traditional Chinese medicine composition, wherein the extract is obtained by co-extracting a mixture of Cyperus rotundus and Gentiana scabra.

[0011] Thirdly, the present invention provides a method for preparing an extract of the lipid-lowering traditional Chinese medicine composition, comprising the following steps: (1) Raw material pretreatment: Take Cyperus rotundus slices and Gentiana scabra slices respectively, crush them to obtain Cyperus rotundus coarse powder and Gentiana scabra coarse powder; (2) Extraction: Take the coarse powder of Cyperus rotundus and coarse powder of Gentiana scabra obtained in step (1) by weight ratio, mix them evenly, extract with solvent, combine the extracts and let stand, take the supernatant, concentrate and dry to obtain the extract of the Chinese herbal composition.

[0012] Preferably, the pulverization in step (1) is pulverized to 10-80 mesh.

[0013] Preferably, in step (2), the solvent is any one or a combination of ethanol and water, and the total mass ratio of Cyperus rotundus powder and Gentiana scabra powder to the volume of solvent is 1:5 to 10, with units of g / ml.

[0014] Preferably, the extraction method is any one or a combination of at least two of the following: soaking, hot reflux extraction, ultrasonic extraction, or decoction, and the number of extractions is 1 to 3.

[0015] Preferably, the extraction is performed by first soaking in 1-2 times the amount of solvent (mass-volume ratio) for 6-8 hours, and then extracting by hot reflux 2-3 times with 3-8 times the amount of solvent (mass-volume ratio), each extraction lasting 1-2 hours.

[0016] Preferably, the settling time in step (2) is 30 to 60 minutes.

[0017] During the co-extraction of Cyperus rotundus and Gentiana scabra, the components in Cyperus rotundus promote the dissolution of active components such as iridoids and terpenes in Gentiana scabra; new active derivatives may be generated under hot, water / alcohol conditions; ineffective or irritating components are removed after precipitating by combining with each other, which helps to improve the activity of the composition and reduce toxic side effects.

[0018] Fourthly, the present invention provides the use of the above-mentioned traditional Chinese medicine composition and extracts thereof in the preparation of medicaments for the prevention and / or treatment of hyperlipidemia.

[0019] Preferably, the drug can improve lipid metabolism disorders in hepatocytes.

[0020] Preferably, the drug can reduce the levels of triglycerides (TG) and total cholesterol (TC) in hepatocytes.

[0021] Preferably, the drug further includes pharmaceutically acceptable excipients; Adding pharmaceutically acceptable excipients to drugs can improve drug stability, increase bioavailability, mask unpleasant odors, improve taste, facilitate formulation processing and storage, thereby improving patient medication adherence and enhancing treatment efficacy. Preferably, the pharmaceutically acceptable pharmaceutical excipient may be selected from one or more of the following: excipients, stabilizers, antioxidants, preservatives, flavoring agents, sweeteners, disintegrants, lubricants, binders, coating materials, sustained-release materials, solubilizers, and pH adjusters.

[0022] Preferably, the dosage form of the drug is an oral liquid, tablet, capsule, granule, or pill.

[0023] In preparing oral liquid dosage forms, extracts of traditional Chinese medicine compositions are mixed with appropriate excipients, and then sterilized and packaged. Oral liquid dosage forms are characterized by rapid onset of action and good absorption, making them particularly suitable for patients with difficulty swallowing.

[0024] In tablet preparation, the extract of a traditional Chinese medicine composition is mixed with appropriate excipients such as starch and microcrystalline cellulose, and then processed through granulation, tableting, and coating. Tablets are convenient to carry and provide accurate dosage, making them one of the most commonly used dosage forms.

[0025] In the preparation of capsules, the extract of the traditional Chinese medicine composition is mixed with an appropriate amount of excipients and filled into gelatin empty capsules. Capsules can mask the special odor of traditional Chinese medicine and improve patient medication compliance.

[0026] In the preparation of granules, the extract of the traditional Chinese medicine composition is mixed with appropriate excipients such as sucrose and sweeteners, and then processed through granulation and drying. Granules are convenient to take, either directly or dissolved in water, and are suitable for all types of people.

[0027] When preparing pills, the extract of the traditional Chinese medicine composition is mixed evenly with an appropriate amount of excipients. This makes them easy to take.

[0028] Compared with the prior art, the present invention has the following significant advantages: 1. Outstanding synergistic effect: This invention is the first to demonstrate through cell model pharmacodynamic experiments that the lipid-lowering effect (reducing TG and TC) of Cyperus rotundus and Gentiana scabra when used together is significantly better than that of the two herbs when used alone, which reflects the synergistic effect under the guidance of the traditional Chinese medicine theory of "soothing the liver and regulating qi" and "clearing heat and purging fire". 2. Significant technological advantages: This invention compares two processes, "mixed extraction" and "separate extraction followed by combination". Experimental data confirms that the lipid-lowering effect of the mixed extract is better, especially the water extract. After the two herbs are co-extracted with water, the lipid-lowering activity of the resulting extract is significantly improved, and cell vitality is enhanced.

[0029] 3. Clear process parameters: Key process parameters such as raw material form (coarse powder), solvent selection (water or 70% ethanol), soaking time, reflux times and time have been optimized and solidified, resulting in good reproducibility and easy standardization and industrial production.

[0030] 4. Clear target and promising application prospects: This invention has been fully validated in a hyperlipidemic hepatocyte injury model (BRL-3A cells), and the mechanism of action is clear (reducing intracellular lipid content), providing a solid experimental basis and candidate material foundation for the development of new traditional Chinese medicines for the treatment of hepatogenic hyperlipidemia and related metabolic diseases. Attached Figure Description

[0031] Figure 1 The results of oil red staining of cell lipid droplets. Detailed Implementation

[0032] The present invention will now be described in further detail with reference to the accompanying drawings and embodiments, but this does not limit the present invention in any way. Any modifications or improvements made based on the teachings of the present invention shall fall within the protection scope of the present invention.

[0033] The processes, conditions, reagents, and experimental methods used in implementing this invention, except as specifically mentioned below, are all common knowledge and general knowledge in the field, and this invention does not have any particular limitations. Experimental methods in the embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer.

[0034] Unless otherwise stated, all technical terms and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. However, in the event of any conflict, the specification containing the definitions shall prevail.

[0035] The following are some of the main raw material sources in the examples: Cyperus rotundus ( Cyperus rotundus L (Source: Yulin, Guangxi; purchased from Yunnan Daodi Chinese Herbal Medicine Pieces Co., Ltd.) Gentian (Gentiana scabra) G. rigescens Franch. The ingredients originated from Lincang, Yunnan, and were purchased from Yunnan Daodi Chinese Herbal Medicine Pieces Co., Ltd. Example 1 A lipid-lowering traditional Chinese medicine composition, which is composed of two traditional Chinese medicinal materials, Cyperus rotundus and Gentiana scabra, with a weight ratio of 2:1 between Cyperus rotundus and Gentiana scabra.

[0036] Example 2 A lipid-lowering traditional Chinese medicine composition, which is composed of two traditional Chinese medicinal materials, Cyperus rotundus and Gentiana scabra, with a weight ratio of 2:2 between Cyperus rotundus and Gentiana scabra.

[0037] Example 3 A lipid-lowering traditional Chinese medicine composition, which is composed of two traditional Chinese medicinal materials, Cyperus rotundus and Gentiana scabra, with a weight ratio of Cyperus rotundus to Gentiana scabra of 2:4.

[0038] Example 4 An extract of a lipid-lowering traditional Chinese medicine composition is prepared by co-extracting two traditional Chinese medicinal materials, Cyperus rotundus and Gentiana scabra, in a 2:2 ratio.

[0039] Its preparation method is as follows: (1) Raw material pretreatment: Take Cyperus rotundus slices and Gentiana scabra slices respectively, crush them and pass them through a 24-mesh sieve to obtain Cyperus rotundus coarse powder and Gentiana scabra coarse powder; (2) Extraction: Take the coarse powder of Cyperus rotundus and coarse powder of Gentiana scabra obtained in step (1) at a weight ratio of 1:1, mix them evenly, add 2 times the amount of water (mass-volume ratio) to soak for 6 hours, add 8 times the amount of water (mass-volume ratio) to extract by hot reflux, extract twice, 1 hour each time, combine the extracts and let stand, take the supernatant, concentrate and dry to obtain the extract of the Chinese herbal composition.

[0040] Example 5 An extract of a lipid-lowering traditional Chinese medicine composition is prepared by co-extracting two traditional Chinese medicinal materials, Cyperus rotundus and Gentiana scabra, in a 2:2 ratio.

[0041] Its preparation method is as follows: (1) Raw material pretreatment: Take Cyperus rotundus slices and Gentiana scabra slices respectively, crush them and pass them through a 24-mesh sieve to obtain Cyperus rotundus coarse powder and Gentiana scabra coarse powder; (2) Extraction: Take the coarse powder of Cyperus rotundus and coarse powder of Gentiana scabra obtained in step (1) at a weight ratio of 1:1, mix them evenly, soak them in 2 times 70% ethanol (mass-volume ratio) for 6 hours, add 8 times 70% ethanol (mass-volume ratio) for hot reflux extraction, extract twice, 1 hour each time, combine the extracts and let them stand, take the supernatant, concentrate and dry to obtain the extract of the Chinese herbal composition.

[0042] Example 6 An extract of a lipid-lowering traditional Chinese medicine composition is prepared by mixing two traditional Chinese medicinal materials, Cyperus rotundus and Gentiana scabra, in a 1:1 ratio and then extracting them together.

[0043] Its preparation method is as follows: (1) Raw material pretreatment: Take Cyperus rotundus slices and Gentiana scabra slices respectively, crush them and pass them through a 24-mesh sieve to obtain Cyperus rotundus coarse powder and Gentiana scabra coarse powder; (2) Extraction: Take the coarse powder of Cyperus rotundus and coarse powder of Gentiana scabra obtained in step (1) at a weight ratio of 1:1, mix them evenly, add 1 times the amount of water (mass-volume ratio) and soak for 8 hours, add 4 times the amount of water (mass-volume ratio) and extract by hot reflux 3 times, 2 hours each time, combine the extracts and let them stand, take the supernatant, concentrate and dry to obtain the extract of the Chinese herbal composition.

[0044] Example 7: Lipid-lowering drug efficacy test (BRL-3A rat hepatocyte hyperlipidemia model) 1. Experimental Materials (1) Cells: BRL-3A rat hepatocytes (purchased from Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd.); (2) Reagents: Oleic acid, palmitic acid (Sigma), TG detection kit, TC detection kit (Nanjing Jiancheng Biotechnology), MTT cell viability detection kit (Beyotime); (3) Test samples: water extract of Cyperus rotundus slices, water extract of Gentiana scabra slices, alcohol extract of Cyperus rotundus slices, alcohol extract of Gentiana scabra slices, 1:1 mixture of water extract of Cyperus rotundus slices and water extract of Gentiana scabra slices, 1:1 mixture of alcohol extract of Cyperus rotundus slices and alcohol extract of Gentiana scabra slices, mixed water extract (prepared in Example 3), mixed alcohol extract (prepared in Example 4).

[0045] 2. Experimental Methods 2.1 Establishment of a hyperlipidemic cell model: BRL-3A cells were cultured to the logarithmic growth phase, and the cell density was adjusted to 5 × 10⁶ cells / year using DMEM medium containing 10% fetal bovine serum. 5Cells were seeded at a density of 1 / mL in 6-well plates (for TG, TC detection and Oil Red O staining) and 96-well plates (for MTT detection). After cell adhesion (12-16 h of culture), the medium was replaced with 2 mmol / L FFA (oleic acid to palmitic acid ratio of 2:1; oleic acid and palmitic acid were dissolved in NaOH and 5% BSA by heating and then diluted with culture medium to the target concentration). The high-lipidemia model was established by culturing for 24 h. The control group was given culture medium containing the same concentration of BSA but without oleic acid / palmitic acid.

[0046] 2.2 Grouping and Dosing: The cells were divided into the following 12 groups (6-well plates n=3, 96-well plates n=10): Control group: Normal BRL-3A cells, supplemented with drug-free culture medium; Model group: high-lipid model cells, supplemented with drug-free culture medium; Cyperus rotundus water extract group: high-lipid model cells were treated with water extract of Cyperus rotundus slices (final concentration 100 μg / mL, preparation method: take 20g of Cyperus rotundus slices and extract using the extraction method and conditions of Example 4). Gentian aqueous extract group: high-lipid model cells were treated with gentian aqueous extract (final concentration 100 μg / mL, preparation method: take 20g of gentian slices and extract using the extraction method and conditions of Example 4). Cyperus rotundus alcohol extract group: high-lipid model cells were treated with ethanol extract of Cyperus rotundus slices (final concentration 100 μg / mL, preparation method: take 20g of Cyperus rotundus slices and extract using the extraction method and conditions of Example 5). Gentian alcohol extract group: high-lipid model cells were treated with gentian alcohol extract (final concentration 100 μg / mL, preparation method: take 20g of gentian slices and extract using the extraction method and conditions of Example 5). Water extraction combination group: high-lipid model cells were added with a 1:1 mixture of water extracts of Cyperus rotundus slices and Gentiana scabra slices (final concentration 100 μg / mL, take 20g of Cyperus rotundus slices and 20g of Gentiana scabra slices, extract them separately with water, and then mix the extracts. The extraction method is the same as in Example 4). Alcohol-extracted combination group: high-lipid model cells were treated with a 1:1 mixture of ethanol extracts of Cyperus rotundus and Gentiana scabra, calculated based on the crude drug (final concentration 100 μg / mL; 20g of Cyperus rotundus and 20g of Gentiana scabra were extracted separately with 70% ethanol, and then the extracts were mixed; the extraction method was the same as in Example 5). Group 1 of mixed water extract (Rhizoma Cyperi to Radix Gentianae 2:1): High-lipid model cells were treated with the mixed water extract from Example 1 (final concentration 100 μg / mL, extraction method same as in Example 4). Two groups of mixed water extracts (Cyperus rotundus and Gentiana scabra in a 2:2 ratio): High-lipid model cells were treated with the mixed water extract from Example 2 (final concentration 100 μg / mL, extraction method as in Example 4). Three groups of mixed water extracts (Rhizoma Cyperi to Radix Gentianae 2:4): High-lipid model cells were treated with the mixed water extract of Example 3 (final concentration 100 μg / mL, extraction method same as Example 4). Mixed alcohol extract group: high-lipid model cells were treated with the mixed alcohol extract from Example 5 (final concentration 100 μg / mL).

[0047] Continue culturing for 24 hours after drug administration, and then detect the indicators using the following methods.

[0048] 2.3 Indicator Testing: Intracellular TG and TC content in hepatocytes: Collect cells from 6-well plates, wash twice with pre-cooled PBS, add cell lysis buffer (containing protease inhibitor), lyse on ice for 30 min, centrifuge at 12000 rpm for 15 min (4℃), and collect the supernatant; follow the kit instructions to detect the concentration of TG and TC in the supernatant, and the results are expressed as "mmol / L"; Cell viability: Add 20 μL of MTT solution (5 mg / mL) to each well of a 96-well plate and continue culturing for 4 h; discard the culture medium, add 150 μL of DMSO to each well, and shake for 10 min to dissolve the crystals; measure the absorbance at 490 nm using a microplate reader (A). 490 Cell viability (%) = (Experimental group A) 490 - Blank Group A 490 ) / (Control Group A) 490 - Blank Group A 490 ) × 100%; Observation of lipid accumulation: 6-well plate cells were washed twice with PBS and fixed with 4% paraformaldehyde for 30 min; Oil Red O staining solution (0.5% Oil Red O in isopropanol solution and distilled water mixed at a ratio of 3:2, filtered and used) was added and stained for 15 min; excess staining solution was washed away with 60% isopropanol and washed twice with PBS; red lipid droplets in the cells were observed under an optical microscope.

[0049] 3. Experimental Results The experimental results are shown in Table 1 and Figure 1 .

[0050] Table 1 Results of lipid-lowering drug efficacy test

[0051] The above results indicate that the proposed Cyperus rotundus-Gentiana combination exhibits a synergistic effect, effectively reversing lipid metabolism disorders and cell damage induced by a high-fat diet. Among these, the mixed water extract group showed the best effect, with significantly stronger lipid-lowering (TG, TC) and cell viability-protecting effects compared to the separately extracted and combined groups and any single herb group.

[0052] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A lipid-lowering traditional Chinese medicine composition, characterized in that, It is composed of Cyperus rotundus and Gentiana scabra, with a weight ratio of 2:1-4.

2. An extract of the lipid-lowering traditional Chinese medicine composition as described in claim 1, characterized in that, The extract was obtained by co-extracting a mixture of Cyperus rotundus and Gentiana scabra.

3. A method for preparing an extract of the lipid-lowering traditional Chinese medicine composition as described in claim 2, characterized in that, Includes the following steps: (1) Raw material pretreatment: Take Cyperus rotundus slices and Gentiana scabra slices respectively, crush them to obtain Cyperus rotundus coarse powder and Gentiana scabra coarse powder; (2) Extraction: Take the coarse powder of Cyperus rotundus and coarse powder of Gentiana scabra obtained in step (1) by weight ratio, mix them evenly, extract with solvent, combine the extracts and let stand, take the supernatant and dry to obtain the extract of the Chinese herbal composition.

4. The preparation method according to claim 3, characterized in that, In step (2), the solvent is any one of ethanol or water or a combination thereof, and the total mass ratio of Cyperus rotundus powder and Gentiana scabra powder to the volume ratio of the solvent is 1:5 to 10, with the unit being g / ml.

5. The preparation method according to claim 3, characterized in that, The extraction method is any one or a combination of at least two of the following: soaking, hot reflux extraction, ultrasonic extraction, or decoction, and the number of extractions is 1 to 3.

6. The use of the traditional Chinese medicine composition of claim 1 and the extract of the traditional Chinese medicine composition of claim 2 in the preparation of a medicament for the prevention and / or treatment of hyperlipidemia.

7. The application according to claim 6, characterized in that, The drug can improve lipid metabolism disorders in hepatocytes.

8. The application according to claim 6, characterized in that, The drug reduces the levels of triglycerides and total cholesterol in liver cells.

9. The application according to claim 6, characterized in that, The drug also includes pharmaceutically acceptable excipients.

10. The application according to claim 6, characterized in that, The dosage form of the drug is oral liquid, tablet, capsule, granule, or pill.

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