A medicinal and food homologous composition for improving metabolic syndrome, and a preparation method and application thereof

Abstract

The application discloses a kind of medicine and food homologous group prescription for improving metabolic syndrome and its preparation method and application, belong to medicine and food homologous functional food technical field, group prescription with Fenugreek, fennel, peony, liquorice, dark plum as raw material, by cleaning, drying, grinding, extraction, filtration, concentration, drying preparation is obtained.The application adopts the above-mentioned one kind of medicine and food homologous group prescription for improving metabolic syndrome and its preparation method and application, and is verified by in vivo high-fat diet induced mouse model and in vitro FFA induced HepG2 cell model, and the group prescription can dose-dependent reduce blood sugar, blood fat, improve liver steatosis, repair mitochondrial damage, inhibit liver cell iron death.The mechanism of action is to regulate lipid metabolism, oxidative stress and iron death process through AKT / Nrf2 / GPX4 signal pathway.

Description

Technical Field

[0001] This invention relates to the field of functional food products that are both food and medicine, and in particular to a formula that improves metabolic syndrome, its preparation method, and its application. Background Technology

[0002] Metabolic syndrome (MS) is a group of metabolic diseases characterized by a cluster of risk factors, including obesity, hyperglycemia, dyslipidemia, and hypertension. Its onset is closely related to obesity, insulin resistance, and abnormal lipid metabolism. Long-term illness can easily induce various complications, seriously affecting human health. Currently, clinical interventions for metabolic syndrome mainly focus on weight loss, dietary adjustments, and regular exercise. There are no specific drugs available, and long-term reliance on Western medicine interventions can easily lead to side effects. Traditional Chinese medicine, however, has unique advantages in the management of metabolic syndrome.

[0003] Although Traditional Chinese Medicine (TCM) does not have a specific disease name for metabolic syndrome, it categorizes it based on its manifestations under the categories of "obesity," "diabetes," "dizziness," and "phlegm turbidity." Scholars generally believe that its occurrence is closely related to deficiencies in the liver, spleen, and kidneys. Spleen deficiency leads to impaired digestion and transformation, resulting in the accumulation of phlegm and dampness; kidney deficiency hinders qi transformation, causing abnormal distribution of body fluids and an accumulation of phlegm and dampness. Over time, this affects the liver's function of regulating qi, leading to qi stagnation and poor blood circulation, further resulting in qi stagnation and blood stasis. Based on this, many TCM compound formulas or formulas containing both medicinal and food ingredients have been explored for the treatment of metabolic syndrome, with some studies showing positive effects such as improving glucose and lipid metabolism and reducing oxidative stress. However, existing TCM intervention programs generally have the following shortcomings: (1) The formulation and compatibility lack sufficient experimental verification, and the pharmacodynamic material basis is unclear; (2) The study of the mechanism of action is mostly limited to the level of apparent indicators, and the regulatory relationship of key signaling pathways is not clearly explained; (3) The preparation process parameters are rough, the quality controllability is poor, and it is difficult to ensure batch-to-batch stability; (4) There is a lack of systematic in vivo and in vitro pharmacodynamic evaluation and safety data, which limits its clinical application and promotion. Summary of the Invention

[0004] The purpose of this invention is to provide a medicinal and food homologous formula for improving metabolic syndrome, its preparation method and application. It improves blood glucose, blood lipids, liver function and lipid metabolism through multi-target synergistic effects, reduces ferroptosis and oxidative stress, and the whole formula follows the treatment principles of harmonizing the liver, spleen and kidneys, removing dampness and turbidity, and nourishing qi and yin. It is suitable for long-term use by people with metabolic syndrome.

[0005] To achieve the above objectives, the present invention provides a medicinal and edible formula for improving metabolic syndrome, comprising the following ingredients by weight: 10-15 parts fenugreek, 6-10 parts fennel, 6-10 parts peony root, 3-5 parts licorice root, and 5-8 parts dried plum.

[0006] Preferably, by weight, it includes the following ingredients: 10 parts fenugreek, 6 parts fennel, 6 parts peony, 3 parts licorice, and 6 parts dried plum.

[0007] Preferably, by weight, it includes the following ingredients: 13 parts fenugreek, 8 parts fennel, 8 parts peony root, 4 parts licorice root, and 7 parts dried plum.

[0008] Preferably, by weight, it includes the following ingredients: 15 parts fenugreek, 10 parts fennel, 10 parts peony root, 5 parts licorice root, and 8 parts dried plum.

[0009] The preparation method of the above-mentioned food-medicine homology formula for improving metabolic syndrome includes the following steps: S1. Clean, dry, pulverize, and sieve the fenugreek, fennel, peony, licorice, and dried plum to obtain powder; S2. Add purified water to the powder, then reflux to extract the extract. Repeat twice. Filter the extracts, combine them, and concentrate under reduced pressure to obtain a concentrated solution. S3. Spray-dry or vacuum-dry the concentrated liquid to obtain the medicinal and edible homologous formula.

[0010] Preferably, in S1, the sieve opening is 20-60 mesh.

[0011] Preferably, in step S2, the amount of purified water used is 10-15 times the volume of the powder obtained in step S1, the reflux extraction temperature is 80-85℃, the reflux extraction time is 1h, the vacuum concentration temperature is 60℃, and the vacuum concentration is carried out to a relative density of 1.10-1.20.

[0012] The above-mentioned medicinal and food homology formula for improving metabolic syndrome is used in the preparation of drugs for treating metabolic syndrome.

[0013] Preferably, pharmaceutically acceptable excipients are added during the preparation of the drug for treating metabolic syndrome, and the resulting drug is one of the following: oral liquid, granules, tablets, or pills.

[0014] Preferably, pharmaceutically acceptable excipients are one or more of dextrin and steviol glycosides, and the dosage of the drug is 6-10g per day, 2-3 times a day.

[0015] Therefore, the present invention, employing the above-mentioned formula for improving metabolic syndrome derived from both food and medicine, its preparation method, and its application, has the following beneficial effects: (1) This invention is based on the traditional formula of fenugreek pills, which is in line with the theory of syndrome differentiation in traditional Chinese medicine. The formula is fixed and has a clear experimental basis. It has been verified by experiments that it can effectively improve the relevant indicators of metabolic syndrome. The core active ingredients that enter the blood are clear and can act on the core targets of metabolic syndrome such as IL6, AKT1, TNF, PPARG, etc., and regulate the AKT / Nrf2 / GPX4 signal axis. It is highly targeted. (2) The present invention uses low-temperature ultrasonic extraction and drying processes, combined with LC-MS technology for component identification, which can accurately extract and retain the core active ingredients such as flavonoids and phenolic acids in the traditional Chinese medicine formula. The extraction efficiency is high, the impurity content is low, and the product efficacy is stable. The process parameters are clear and the repeatability is strong. (3) All raw materials used in this invention are natural Chinese medicines that meet the pharmacopoeia standards. Experiments have verified that there are no obvious adverse reactions, and the safety is high. They are suitable for long-term conditioning of people with metabolic syndrome. The dosage form is pills, which are convenient to take and can improve patient compliance. (4) Based on existing experimental research results, this invention clarifies the pharmacodynamic material basis, target and regulatory pathway of traditional Chinese medicine formulation. The preparation process is replicable and easy to promote, and can provide a clear technical solution for the treatment of metabolic syndrome with traditional Chinese medicine, and guide the rational clinical application.

[0016] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0017] Figure 1 This is a positive ion diagram for the identification of traditional Chinese medicine components in pills prepared in Example 3 of this invention; Figure 2 This is a negative ion diagram for the identification of traditional Chinese medicine components in the pills prepared in Example 3 of this invention; Figure 3 This is a pie chart showing the classification of Chinese herbal components in the pills prepared according to Example 3 of this invention; Figure 4 This is a distribution diagram of the number of traditional Chinese medicine components in the pills prepared according to Example 3 of the present invention; Figure 5 This is a positive ion diagram for the identification of blood-entering components of the pills prepared in Example 3 of this invention; Figure 6 This is a negative ion diagram for the blood-entry components of the pills prepared in Example 3 of this invention; Figure 7 This is a blood-entry component classification and content distribution diagram of the pills prepared in Example 3 of this invention; Figure 8 This is a distribution diagram of the number of blood-entering components in the pills prepared in Example 3 of this invention; Figure 9 These are illustrations showing the changes in body weight, blood glucose, blood lipids, and liver index in mice on high-fat diets in each group according to the present invention. Figure 10 This is a schematic diagram of the HE, Oil Red O, and Masson staining results of high-fat diet mice in each group according to the present invention; Figure 11 These are transmission electron microscope images of the mitochondrial ultrastructure repair in mice on high-fat diets according to this invention. Figure 12This is a schematic diagram of the Western blot detection results of the present invention; Figure 13 These are the results of CCK8 assay in HepG2 cells of mice on a high-fat diet in each group according to this invention; Figure 14 These are the Oil Red O staining results of HepG2 cells from high-fat diet mice in each group according to this invention; Figure 15 This refers to the intracellular TG content levels in HepG2 cells of high-fat diet mice in each group of the present invention. Figure 16 This refers to the intracellular ROS levels in HepG2 cells of high-fat diet mice in each group of the present invention. Figure 17 This invention presents the results of flow cytometry analysis of ROS levels in HepG2 cells of mice on a high-fat diet in each group. Figure 18 This invention relates to the detection of mitochondrial membrane potential levels in cells of mice on a high-fat diet using staining confocal microscopy. Figure 19 This invention presents the results of flow cytometry JC-1 detection of mitochondrial membrane potential in mice on high-fat diets in each group. Detailed Implementation

[0018] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention should be considered equivalent substitutions and are included within the protection scope of the present invention. Furthermore, it should be understood that after reading the contents of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims and are all within the protection scope of the present invention.

[0019] In this document, the term "embodiment" means that a specific feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of this application. The term "embodiment" appearing in various places throughout the specification does not necessarily refer to the same embodiment, nor does it specifically limit its independence or connection with other embodiments. In principle, in this application, as long as there are no technical contradictions or conflicts, the technical features mentioned in each embodiment can be combined in any way to form corresponding implementable technical solutions.

[0020] Unless otherwise defined, the technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains; the use of related terms herein is merely for the purpose of describing particular embodiments and is not intended to limit this application.

[0021] In this invention, unless otherwise specified, all other test materials and instruments are conventional test materials in the field and can be purchased through commercial channels.

[0022] Example 1 This invention provides a medicinal and edible formula for improving metabolic syndrome, comprising the following ingredients by weight: 10 parts fenugreek, 6 parts fennel, 6 parts peony root, 3 parts licorice root, and 6 parts dried plum.

[0023] The preparation method of the above-mentioned food-medicine homology formula for improving metabolic syndrome includes the following steps: S1. Clean, dry, and pulverize fenugreek, fennel, peony, licorice, and dried plum to obtain powder; S2. Add 12 times the volume of purified water to the powder, then reflux at 85°C for 1 hour to obtain the extract. Repeat twice. Filter the extracts and combine them. Concentrate under reduced pressure at 60°C to a relative density of 1.15 to obtain the concentrate. S3. Spray-dry or vacuum-dry the concentrated liquid to obtain the medicinal and edible homologous formula.

[0024] The prepared food-medicine homology formula was used to prepare drugs for the treatment of metabolic syndrome, fatty liver and NAFLD; dextrin and steviol glycosides were added to make pills when preparing drugs for the treatment of metabolic syndrome, fatty liver and NAFLD, and the pills were denoted as HLBW-L (abbreviated as L).

[0025] Example 2 The only difference between this embodiment and Embodiment 1 is that, by weight, it includes the following ingredients: 13 parts fenugreek, 8 parts fennel, 8 parts peony root, 4 parts licorice root, and 7 parts dried plum. All other conditions are the same, and the final pill is denoted as HLBW-M (abbreviated as M).

[0026] Example 3 The only difference between this embodiment and Embodiment 1 is that, by weight, it includes the following ingredients: 15 parts fenugreek, 10 parts fennel, 10 parts peony root, 5 parts licorice root, and 8 parts dried plum. All other conditions are the same, and the final pill is denoted as HLBW-H (abbreviated as H).

[0027] Figure 1-8 The figures show the positive and negative ion chromatograms, classification of Chinese medicine components, and distribution of the number of Chinese medicine components in the pills prepared in Example 3. As can be seen from the figures, the positive and negative ion chromatograms of the Chinese medicine components show good chromatographic peak separation and stable response intensity.

[0028] Using LC-MS technology combined with comparative analysis of MJBIOTCM (MJ BiotCM's proprietary database of metabolites of traditional Chinese medicine), 652 compounds were preliminarily identified in the pills, covering 11 major categories including flavonoids, terpenes, phenolic acids and their derivatives. Among them, flavonoids accounted for the highest percentage of substances (25.78%) and 40.99% of the total content; followed by phenolic acids and their derivatives and terpenes.

[0029] In Example 3, 112 active ingredients of the pills were identified that could be absorbed into the bloodstream. The screening criteria for blood-absorbing ingredients were as follows: 1. Substances common to both the traditional Chinese medicine and the serum used for the medicine, but not found in the blank serum; 2. Substances contained in the traditional Chinese medicine, and whose content in the serum used for the medicine was twice that in the blank serum. If either criterion was met, the ingredient was determined to be a blood-absorbing ingredient. Among the blood-absorbing ingredients, in terms of the percentage of substances, flavonoids accounted for the highest proportion, reaching 48.68%, making them the dominant blood-absorbing ingredients; followed by phenolic acids and their derivatives (12.5%) and terpenes (11.84%). In terms of the percentage of content, lipids accounted for the highest proportion (51.12%), followed by alkaloids and their derivatives (11.72%) and phenolic acids and their derivatives (11.13%). Venn diagram of blood-entering components showed that a total of 652 components were identified in the herb sample, 10 components were identified in the blank serum sample, and 83 components were identified in the treatment serum sample. Among them, there were 112 components in the herb and treatment serum samples, which are potential active blood-entering components. Effect verification: A metabolic syndrome model was constructed using SPF-grade C57BL / KsJ-Leprdb / db (db mice). Simultaneously, littermate negative C57BL / KsJ-Leprwt / wt (wt mice) were selected as the normal control group. After successful model construction, db mice were randomly divided into experimental groups, positive control group, model control group, and normal control group. The experimental groups included HLBW-L, HLBW-M, and HLBW-H groups. There were no significant differences in age and weight among the groups (P>0.05), making them comparable.

[0030] Medication usage details: Experimental group: HLBW-L group: The pills prepared in Example 1 of this invention were administered by gavage at a dose of 2.3 g / kg / d once a day for 49 consecutive days. HLBW-M group: The pills prepared in Example 2 of this invention were administered by gavage at a dose of 4.55 g / kg / d once a day for 49 consecutive days. HLBW-H group: The pills prepared in Example 3 of this invention were administered by gavage at a dose of 9.1 g / kg / d once a day for 49 consecutive days. Positive control group: Administered commercially available metformin via gavage at a standard dose of 0.2275 g / kg / day once daily for 49 consecutive days; Model control group: The same volume of distilled water as the experimental group was administered by gavage once a day for 49 consecutive days. Normal control group: Administered an equal volume of distilled water by gavage once daily for 49 consecutive days.

[0031] During the experiment, all groups of mice maintained a normal diet and regular routine. The model control group, experimental group, and positive control group were fed a high-fat diet, while the normal control group was fed a normal diet.

[0032] Fasting blood glucose, liver tissue TC, TG, AST, and ALT levels were measured in mice before and after the experiment. Changes in mouse body weight and pathological morphology of adipose tissue were observed. The expression levels of core proteins such as AKT, Nrf2, and GPX4 in adipose tissue and liver tissue were measured. Adverse reactions in mice were recorded.

[0033] The results are as follows: Results of in vivo mouse experiments are as follows Figure 9 As shown, compared with the model control group, the experimental group mice had significantly lower body weight, significantly lower fasting blood glucose, serum TC and TG levels (P<0.05), significantly lower liver function indicators AST and ALT levels, and improved liver index. Results of HE staining, Oil Red O staining, and Masson staining are as follows: Figure 10 As shown, by Figure 10 It can be seen that the experimental group of mice showed a significant reduction in liver fat deposition, liver tissue inflammation infiltration, and collagen deposition; Transmission electron microscopy observations as follows Figure 11 As shown, by Figure 11 It can be seen that the ultrastructure of mitochondria in the liver cells of mice in the experimental group was repaired, the mitochondrial volume returned to normal, the outer membrane was intact, the cristae were neatly arranged, and the mitochondrial damage was significantly alleviated.

[0034] Western blot results are as follows Figure 12 As shown, by Figure 12 It was found that the expression of p-AKT, Nrf2, HO-1, and GPX4 proteins was significantly upregulated in the experimental group cells (P<0.05), the expression of HDAC3 protein was significantly downregulated (P<0.05), and the expression of SLC7A11 protein was upregulated, suggesting that hepatocyte ferroptosis was inhibited.

[0035] Figure 13The results of CCK8 assays in HepG2 cells from each group are presented. The CCK8 assay is used to detect cell viability or proliferation. In the FFA (free fatty acid)-induced HepG2 cell model, cell viability in the model control group was significantly decreased, while the experimental groups given different doses of the pills (HLBW-L, HLBW-M, HLBW-H) showed a dose-dependent increase in hepatocyte viability, indicating that this food-medicine homology formula has a protective effect against FFA-induced hepatocyte damage.

[0036] Figure 14 The Oil Red O staining results of HepG2 cells in each group are shown. Oil Red O staining is used to observe intracellular lipid deposition. The results showed that after FFA induction, intracellular lipid deposition in the model control group increased significantly; while after treatment with the pills, lipid deposition in the experimental group was significantly reduced, indicating that the formula can effectively inhibit lipid accumulation in hepatocytes and improve steatosis.

[0037] Figure 15 The study presented the intracellular TG (triglyceride) levels in HepG2 cells from each group. Consistent with Oil Red O staining results, the TG levels in the model control group cells were significantly increased; while the TG levels in the experimental group cells decreased in a dose-dependent manner, further validating that the pills can reduce triglyceride levels in hepatocytes, thereby improving lipid metabolism disorders.

[0038] In vitro cell experiment results as follows Figures 16-17 As shown, by Figure 6 It can be seen that in the FFA-induced HepG2 cell model, the experimental group can significantly restore the mitochondrial membrane potential of hepatocytes, reduce intracellular ROS generation and lipid deposition, and enhance hepatocyte viability. Mechanism verification results as follows Figures 18-19 As shown in the figure, the experimental group can regulate lipid metabolism, improve oxidative stress, and inhibit hepatocyte ferroptosis by activating the AKT / Nrf2 / GPX4 signaling pathway, thereby improving the pathological state related to metabolic syndrome, alleviating hepatic lipid deposition and mitochondrial damage, and enhancing the liver's antioxidant capacity.

[0039] Safety results: No obvious adverse reactions were observed in the experimental group and the normal control group mice, and their diet and activity were normal. The model control group mice showed symptoms such as abnormal weight gain, glucose and lipid metabolism disorders, and liver enlargement. Some mice in the positive control group experienced gastrointestinal discomfort. This indicates that the traditional Chinese medicine formula of this invention is safe and has no obvious toxic side effects, and is suitable for long-term use.

[0040] In summary, the food-medicine homology formula of the present invention can improve metabolic syndrome-related indicators in a dose-dependent manner, reduce blood glucose and blood lipids, improve liver fat deposition and mitochondrial damage, regulate lipid metabolism and oxidative stress and inhibit ferroptosis through the AKT / Nrf2 / GPX4 signaling pathway, and is safe and reliable. It can be used for the prevention and adjunctive treatment of metabolic syndrome and related fatty liver.

[0041] Therefore, this invention employs the aforementioned medicinal and edible homologous formula for improving metabolic syndrome, along with its preparation method and application. Verification was conducted using an in vivo high-fat diet-induced mouse model and an in vitro FFA-induced HepG2 cell model. The formula effectively reduces blood glucose and lipids in a dose-dependent manner, improves hepatic steatosis, repairs mitochondrial damage, and inhibits hepatocyte ferroptosis. Its mechanism of action involves regulating lipid metabolism, oxidative stress, and ferroptosis through the AKT / Nrf2 / GPX4 signaling pathway.

[0042] 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 preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A medicinal and edible formula for improving metabolic syndrome, characterized in that: By weight, it includes the following ingredients: 10-15 parts fenugreek, 6-10 parts fennel, 6-10 parts peony root, 3-5 parts licorice root, and 5-8 parts dried plum.

2. The medicinal and edible formula for improving metabolic syndrome according to claim 1, characterized in that: By weight, it includes the following ingredients: 10 parts fenugreek, 6 parts fennel, 6 parts peony root, 3 parts licorice root, and 6 parts dried plum.

3. The medicinal and edible homologous formula for improving metabolic syndrome according to claim 1, characterized in that: By weight, it includes the following ingredients: 13 parts fenugreek, 8 parts fennel, 8 parts peony root, 4 parts licorice root, and 7 parts dried plum.

4. A medicinal and edible formula for improving metabolic syndrome according to claim 1, characterized in that: By weight, it includes the following ingredients: 15 parts fenugreek, 10 parts fennel, 10 parts peony root, 5 parts licorice root, and 8 parts dried plum.

5. A method for preparing a medicinal and edible homologous formula for improving metabolic syndrome as described in any one of claims 1-4, characterized in that: Includes the following steps: S1. Clean, dry, pulverize, and sieve the fenugreek, fennel, peony, licorice, and dried plum to obtain powder; S2. Add purified water to the powder, then reflux to extract the extract. Repeat twice. Filter the extracts, combine them, and concentrate under reduced pressure to obtain a concentrated solution. S3. Spray-dry or vacuum-dry the concentrated liquid to obtain the medicinal and edible homologous formula.

6. The method for preparing a medicinal and edible homologous formula for improving metabolic syndrome according to claim 5, characterized in that: In S1, the sieve opening is 20-60 mesh.

7. The method for preparing a medicinal and edible homologous formula for improving metabolic syndrome according to claim 5, characterized in that: In S2, the amount of purified water used is 10-15 times the volume of the powder obtained in S1, the reflux extraction temperature is 80-85℃, the reflux extraction time is 1h, the vacuum concentration temperature is 60℃, and the vacuum concentration is carried out to a relative density of 1.10-1.

20.

8. The use of a medicinal and food homologous formula for improving metabolic syndrome as described in any one of claims 1-4 in the preparation of a drug for treating metabolic syndrome.

9. The application of a medicinal and edible homology formula for improving metabolic syndrome according to claim 8, characterized in that: Pharmaceutically acceptable excipients are added during the preparation of drugs for treating metabolic syndrome, and the resulting drugs are one of the following: oral liquid, granules, tablets, or pills.

10. The application of a medicinal and edible homology formula for improving metabolic syndrome according to claim 8, characterized in that: Pharmaceutically acceptable excipients include one or more of dextrin and steviol glycosides. The dosage of the drug is 6-10g per day, 2-3 times a day.

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