Compound lipid-lowering tea and preparation method thereof

Abstract

The application discloses the technical field of lipid-lowering tea, and discloses a compound lipid-lowering tea and a preparation method thereof.The compound lipid-lowering tea comprises 10g of cassia seed, 5-15g of gynostemma pentaphyllum, 5-15g of salvia miltiorrhiza, 5-15g of hawthorn and 5-15g of bidens pilosa.The lipid-lowering active ingredients in medicinal materials, such as tanshinone, flavone, ursolic acid and anthraquinone, are studied, and it is clear that these ingredients play a lipid-lowering role through multiple signal pathways, such as the PI3K-AKT signal pathway, the cancer pathogenesis pathway and the estrogen signal pathway.Among them, the flavone is combined with the core target point PPARG, the anthraquinone is combined with HSP90AA, the ursolic acid is combined with ESR1, the lipid synthesis and metabolism are regulated, the fat index, the serum low-density lipoprotein and the triglyceride level are reduced, and the synthesis of high-density lipoprotein is promoted.The animal experiment verifies that the lipid-lowering tea has a significant lipid-lowering effect, and has no adverse effect on the organs of mice, thereby providing a new safe and effective lipid-lowering option for patients with hyperlipidemia.

Description

Technical Field

[0001] This invention relates to the field of lipid-lowering tea technology, specifically to a compound lipid-lowering tea and its preparation method. Background Technology

[0002] Hyperlipidemia, commonly known as high blood lipids or dyslipidemia, is generally defined as elevated levels of triglycerides (TG) and / or total cholesterol (TC) in blood plasma. It also includes elevated low-density lipoprotein cholesterol (LDL-C) and decreased high-density lipoprotein cholesterol (HDL-C). According to the World Health Organization, more than 2 billion people worldwide have hyperlipidemia, and this number continues to grow. Hyperlipidemia can lead to atherosclerosis, coronary heart disease, stroke, and other diseases, and also increases the risk of diabetes, hypertension, and other illnesses, posing a serious threat to human health. Therefore, the prevention and treatment of hyperlipidemia are of great significance for improving human health and promoting economic development.

[0003] Currently, commonly used drugs for treating hyperlipidemia include statins, fibrates, niacin, bile acid sequestrants, cholesterol absorption inhibitors, and PCSK9 inhibitors. However, Western medicine has the following drawbacks: 1. Drug addiction and dependence: Once patients start taking Western medicine to treat hyperlipidemia, they must adhere to long-term use, otherwise blood lipids will rebound. Therefore, patients may develop drug addiction and dependence. 2. Drug complications: Western medicine treatment for hyperlipidemia has certain side effects, such as causing muscle damage, leading to rhabdomyolysis, or causing liver and kidney damage, intestinal discomfort, etc. 3. High treatment costs: Lipid-lowering Western medicine is not affordable and requires long-term use, placing a certain financial burden on patients. 4. Limited treatment effects: Some patients do not respond well to Western medicine treatment for hyperlipidemia, especially the elderly, whose adverse drug reactions are more prominent and difficult to tolerate. 5. Negative health effects: Long-term use of Western medicine to treat hyperlipidemia may adversely affect the patient's immune system and increase the risk of other diseases. Therefore, finding a lipid-lowering drug that is effective, low-cost, and safe is urgent. Summary of the Invention

[0004] The present invention aims to provide a method for preparing and applying a compound lipid-lowering tea, in order to solve the problems of addiction, complications and low safety of existing drugs for treating hyperlipidemia.

[0005] To solve the above-mentioned technical problems, the present invention provides the following technical solution: a compound lipid-lowering tea, comprising 10g of cassia seed, 5-15g of gynostemma pentaphyllum, 5-15g of salvia miltiorrhiza, 5-15g of hawthorn, and 5-15g of beggar-ticks.

[0006] Furthermore, the ingredients are 10g of cassia seed, 5g of gynostemma pentaphyllum, 5g of salvia miltiorrhiza, 5g of hawthorn, and 5g of beggar-ticks.

[0007] Furthermore, the ingredients are 10g of Cassia Seed, 5g of Gynostemma pentaphyllum, 10g of Salvia miltiorrhiza, 10g of Crataegus pinnatifida, and 10g of Bidens pilosa.

[0008] Furthermore, the ingredients are 10g of Cassia Seed, 5g of Gynostemma pentaphyllum, 15g of Salvia miltiorrhiza, 15g of Crataegus pinnatifida, and 15g of Bidens pilosa.

[0009] Furthermore, the ingredients are 10g of Cassia Seed, 10g of Gynostemma pentaphyllum, 5g of Salvia miltiorrhiza, 10g of Crataegus pinnatifida, and 15g of Bidens pilosa.

[0010] Furthermore, the ingredients are 10g of Cassia Seed, 15g of Gynostemma pentaphyllum, 15g of Salvia miltiorrhiza, 10g of Crataegus pinnatifida, and 5g of Bidens pilosa.

[0011] Furthermore, the ingredients are 10g of Cassia Seed, 10g of Gynostemma pentaphyllum, 15g of Salvia miltiorrhiza, 5g of Crataegus pinnatifida, and 10g of Bidens pilosa.

[0012] Furthermore, the ingredients are 10g of Cassia Seed, 15g of Gynostemma pentaphyllum, 10g of Salvia miltiorrhiza, 5g of Crataegus pinnatifida, and 15g of Bidens pilosa.

[0013] Furthermore, the preparation method of the compound lipid-lowering tea according to any one of the above includes the following steps:

[0014] S1. Pretreatment: Clean and dry each Chinese herb, then grind it into powder using a Chinese herb grinder.

[0015] S2. Weighing: Weigh the required weights of cassia seeds, gynostemma pentaphyllum, salvia miltiorrhiza, hawthorn, and beggar-ticks using a weighing device, and soak the mixed powder in an appropriate amount of water for a period of time.

[0016] S3. Boil the soaking liquid in S2 over high heat for a period of time, then boil the soaking liquid over low heat for a period of time.

[0017] S4. Filter the decoction in S3 using gauze and collect the filtrate.

[0018] S5. Add an appropriate amount of water to the filtered dregs and boil over high heat for a period of time, then boil over low heat for a period of time.

[0019] S6. Filter the decoction in S5 using gauze and combine the filtrates;

[0020] S7. The filtrate is concentrated in a water bath to obtain the compound lipid-lowering tea.

[0021] Further, in step S2, the mixed powder is soaked in 1L of purified water for half an hour; in step S2, the soaking liquid is first boiled over high heat for 30 minutes, and then boiled over low heat for 1 hour; in step S5, the filtered residue is added to 0.5L of purified water and boiled over high heat for 30 minutes, and then boiled over low heat for 1 hour; in step S7, the filtrate is concentrated to 100mL in a 100℃ water bath to obtain the compound lipid-lowering tea.

[0022] The beneficial effects of this invention are as follows: 1. Through network pharmacology analysis and molecular docking technology, the main lipid-lowering active ingredients (tanshinone, flavonoids, ursolic acid, and anthraquinones) and their mechanisms of action in the compound lipid-lowering tea were clearly identified. These active ingredients can exert lipid-lowering effects through multiple pathways and targets, such as inhibiting fatty acid synthase activity, activating PPARs to enhance fatty acid oxidation metabolism, regulating intestinal microbiota to affect lipid absorption and metabolism, anti-oxidation to reduce ox-LDL levels, and affecting bile acid metabolism and excretion. The binding relationships with core targets (PPAGR, HSP90AA, ESR1, etc.) were also precisely determined. This not only provides a solid theoretical basis for the lipid-lowering effect of the tea but also lays the foundation for further in-depth research and formulation optimization, contributing to the development of the field of lipid-lowering traditional Chinese medicine. 2. Through animal experiments, the lipid-lowering effect of the compound lipid-lowering tea was verified from multiple aspects, including body weight, organ index, four blood lipid indicators, and liver histopathology. Experimental results show that the lipid-lowering tea not only significantly reduces the body weight of mice, but also regulates four blood lipid indicators, reduces fat formation in liver cells, and improves the pathological characteristics of liver tissue. Simultaneously, it has a positive effect on kidney indices, improving kidney atrophy caused by a high-fat diet, and has no adverse effects on other organs. This fully demonstrates the effectiveness and safety of the lipid-lowering tea in vivo, increasing the product's credibility and application value. 3. The compound lipid-lowering tea of ​​this invention uses cassia seed, gynostemma pentaphyllum, salvia miltiorrhiza, hawthorn, and beggar-ticks as its formula, and clearly defines the dosage range of each component. Compared with other comparative documents, this formula combination and ratio are unique. This unique formula has been experimentally verified to precisely target hyperlipidemia. In animal experiments, the lipid-lowering tea group showed a significantly reduced fat index, with better effects than simvastatin, effectively accelerating fat decomposition and reducing accumulation in hyperlipidemic mice, demonstrating good lipid-lowering efficacy, and providing a new safe and effective option for patients with hyperlipidemia. 4. Most of the medicinal materials in the formula are food and medicine homologous, ensuring safety for long-term consumption. Animal experiments showed that, compared with the normal control group, the lipid-lowering tea group had no significant adverse effects on the organ indices of mice, such as liver, spleen, heart, testis, and epididymis (p>0.05), indicating that it would not damage the growth and development of these important organs, reducing the risk of liver and kidney damage that may be caused by traditional lipid-lowering Western medicines, and improving the safety of users. Attached Figure Description

[0023] Figure 1A diagram showing the common target genes of compound lipid-lowering tea and hyperlipidemia;

[0024] Figure 2 A is a diagram of the intersection of gene-protein interaction networks;

[0025] Figure 2 B is a diagram of the drug-component-target-disease interaction PPI network.

[0026] Figure 3 These are the core targets of the top 8 PPI networks in degree ranking;

[0027] Figure 4 A represents the GO functional enrichment analysis results of the common target genes;

[0028] Figure 4 B represents the enrichment analysis results of the KEGG signaling pathway, a common target gene.

[0029] Figure 5 The 3D structures of the core active ingredients are: A: 3D structure of anthraquinone, B: 3D structure of ursolic acid, C: 3D structure of tanshinone, and D: 3D structure of flavonoids.

[0030] Figure 6 This refers to the binding ability of the core active ingredient to the core protein;

[0031] Figure 7 The docking modes of target molecules and small molecule ligands are as follows: A: docking mode of ERS1 target with hawthorn ursolic acid molecule; B: docking mode of HSP90AA1 target with cassia seed anthraquinone molecule; C: docking mode of PPARG target with flavonoid molecule.

[0032] Figure 8 The trend of mouse body weight change;

[0033] Figure 9 The effect of lipid-lowering tea on organ indices in mice;

[0034] Figure 10 The results of four lipid assays in mouse serum;

[0035] Figure 11 This is the Oil Red O staining procedure;

[0036] Figure 12 Oil Red O staining results of liver tissue from mice in each group: A: normal control group, B: high-fat model group, C: simvastatin group, D: lipid-lowering drug group;

[0037] Figure 13 HE staining steps;

[0038] Figure 14HE staining results of liver tissues from mice in each group: A: normal control group, B: hyperlipidemia model group, C: simvastatin group, D: lipid-lowering tea group. Detailed Implementation

[0039] The following detailed description illustrates the specific implementation method:

[0040] Example:

[0041] This invention selects hawthorn, gynostemma pentaphyllum, cassia seed, salvia miltiorrhiza, and beggar-ticks to prepare a compound lipid-lowering tea in different proportions. Network pharmacology methods are used to analyze the active ingredients of these five traditional Chinese medicines and their lipid-lowering molecular networks, providing a preliminary understanding of the lipid-lowering mechanism of the compound lipid-lowering tea. Volunteers are then recruited to rate the taste, color, and other aspects of the different proportions of the lipid-lowering tea. The formula with the highest score is selected for animal experiments. The lipid-lowering effect and mechanism of action of this formula are explored from four indicators: body weight, organ index, blood lipids, and pathology. This provides a compound lipid-lowering tea with good taste, high safety, and good efficacy for people with high blood lipids, which can be taken in daily life to achieve the purpose of lowering blood lipids. It also provides a scientific basis for the development and promotion of traditional Chinese medicine theory.

[0042] 1. Experimental Materials

[0043] 1.1 Experimental reagents

[0044] Cassia seed, Salvia miltiorrhiza, Gynostemma pentaphyllum, hawthorn, Bidens pilosa, simvastatin.

[0045] 1.2 Laboratory Animals

[0046] This experiment used Kunming mice, SPF grade, male, weighing 18-20g. The experimental animals were housed in an air-conditioned environment with natural light, a temperature of 23-25℃, and humidity of 40%-60%. All mice had free access to food and water.

[0047] 1.3 Main Reagents and Instruments

[0048] Serum biochemical assay kits: TC assay kit, TG assay kit, HDL-C assay kit, LDL-C assay kit. TGL-16 centrifuge; ACS electronic balance; spectrophotometer.

[0049] 2. Experimental Methods

[0050] 2.1 Network pharmacology analysis of the active ingredients of the compound lipid-lowering tea and its common target genes with hyperlipidemia.

[0051] Drug component collection: Literature review was conducted to collect and organize the names of active chemical components with lipid-lowering effects in each traditional Chinese medicine ingredient. All component names were searched in the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP, https: / / tcmspw.com / ). With oral bioavailability (OB) ≥30% and drug-likeness (DL) ≥0.18 as thresholds, all detected chemical components were screened to obtain the potential effective components in the compound lipid-lowering tea.

[0052] Target Collection: To determine the intersection between drug component targets and disease gene targets, tanshinone I, flavonoids, ursolic acid, and anthraquinone were collected as active ingredients in the compound traditional Chinese medicine lipid-lowering tea for the effective treatment of hyperlipidemia. These active ingredients can exert lipid-lowering effects through multiple pathways and targets. For example, they can reduce fatty acid synthesis by inhibiting the activity of fatty acid synthases and other key enzymes; enhance fatty acid oxidation and metabolism and promote lipid transport by activating peroxisome proliferator-activated receptors (PPARs); regulate the gut microbiota, thereby affecting lipid absorption and metabolism; possess antioxidant effects that can reduce oxidative stress and lower oxidized low-density lipoprotein cholesterol (ox-LDL) levels in the blood; and alter lipid digestion and absorption by affecting bile acid metabolism and excretion. These effects may be interrelated and collectively regulate blood lipid levels.

[0053] SMILES numbers for four components were collected from the Pubchem database. Using the collected results, targets with a probability > 0.1 were searched in the Swisstargetprediction database. The intersection of hyperlipidemia-related targets and the effective active ingredients of the compound traditional Chinese medicine lipid-lowering tea was obtained from the GeneCards database, and a Venn diagram was plotted (e.g., ...). Figure 1 (As shown). By Figure 1 It was found that a total of 1,300 disease targets were screened for hyperlipidemia, and 115 of the targets were found in the four active ingredients of traditional Chinese medicine. Among them, 49 targets were common to both, accounting for 3.3% of all targets. These 49 common target genes provide the molecular basis for the lipid-lowering effect of lipid-lowering drugs.

[0054] The 49 targets that intersect with hyperlipidemia and drugs were uploaded to the STRING database to obtain an interaction network diagram. Figure 2 A), then use Cytoscape 3.7.2 to draw the PPI network ( Figure 2 B). Core gene analysis was performed using the cytoNCA plugin, and the top 8 genes by degree were PPAGR, ESR1, HSP90AA1, PTGS2, EGFR, CASP3, BCL2, and MMP9. Figure 3 The above-mentioned target genes may be the core targets of compound lipid-lowering tea in treating hyperlipidemia. Among them, peroxisome proliferator-activated receptor (PPAGR) can promote the uptake and storage of fatty acids, increase adipocyte differentiation, thereby reducing plasma lipid levels, and can regulate the expression of transport proteins and lipases related to lipid metabolism, thus affecting lipid metabolism. Therefore, PPAGR is an important target for the lipid-lowering effect of lipid-lowering tea.

[0055] GO function and KEGG signaling pathway enrichment analysis: The DAVID platform was used to perform GO function enrichment analysis on potential targets of traditional Chinese medicine compound lipid-lowering tea in the treatment of hyperlipidemia. Figure 4 A shows that the biological process (BP) category mainly includes cellular responses to hormones, organic cyclic compounds, and steroid hormones; the cellular component (CC) category mainly includes membrane rafts, membrane microdomains, plasma membrane rafts, and myelin sheaths; and the molecular function (MF) category mainly involves nuclear receptor activity, ligand-activated transcription factor activity, steroid binding, estrogen-responsive element binding, and transcription factor binding. To further understand the specific mechanism by which lipid-lowering tea lowers blood lipids, KEGG signaling pathway enrichment analysis was performed on 49 targets. The results showed significant enrichment in pathways such as the PI3K-AKT signaling pathway, cancer pathogenesis pathway, estrogen signaling pathway, endocrine resistance, and tyrosine kinase inhibitor resistance. Figure 4 (B) This indicates that the active ingredients in lipid-lowering tea mainly exert their lipid-lowering effects by intervening in these signaling pathways. The PI3K-Akt signaling pathway is significantly linked to multiple lipid metabolism-related genes, multiple lipid metabolism organs, and various lipid metabolism disorders. This signaling pathway directly or indirectly participates in the molecular mechanisms of lipid synthesis, transport, uptake, and breakdown, as well as signal transduction regulation under pathological conditions. The active ingredients in lipid-lowering drugs can regulate blood lipid levels and exert their lipid-lowering effects by intervening in the PI3K-Akt signaling pathway.

[0056] 2.2 Molecular docking

[0057] After processing with Chem3D software, the 3D structures of compounds Anthraquinone, Ursolicacid, Tanshinone I, and Flavonoid in the energy minimization scheme are shown below. Figure 5 The first docking of three small molecule ligands with protein receptor genes was performed using AutoDockVina software, and the resulting schematic diagram and binding energy table were compiled. Figure 6The results showed that no hydrogen bonds existed in the binding modes of ESR1 with Flavonoid and ESR1 with Anthraquinone, so these results were discarded. The second docking of the results for PPARG with Flavonoid, ESR1 with Ursolic acid, and HSP90AA1 with Anthraquinone yielded the names of the respective binding site residues and hydrogen bond energies, as shown in [link to documentation]. Figure 7 Analysis revealed that PPARG and Flavonoid require the lowest binding energy, exhibiting the best binding affinity between the small molecule ligand and the protein receptor, resulting in more stable binding.

[0058] 2.3 Preparation of Compound Lipid-Lowering Tea

[0059] Using a daily dosage of 10g of cassia seed as the standard, a four-factor (hawthorn, gynostemma pentaphyllum, salvia miltiorrhiza, and beggar-ticks) three-level (5, 10, 15g) orthogonal array was designed to determine nine different proportions of lipid-lowering tea, as shown in Table 1. The lipid-lowering tea was prepared according to the proportions. First, all Chinese herbs were washed and dried, then pulverized into powder using a Chinese herbal medicine grinder. The dosage of each herb was weighed according to the proportions, and each herb was soaked in purified water for half an hour. The mixture was first boiled over high heat for 30 minutes, then simmered over low heat for 1 hour. The filtrate was filtered and concentrated to 100mL in a water bath. One hundred volunteers were recruited to taste the lipid-lowering tea and rate its color (5 points), aroma (5 points), and taste (5 points), with taste and aroma accounting for 40% and color accounting for 20%. The four lipid-lowering teas with the highest scores were selected. Then, another 117 volunteers were recruited to rate the four highest-scoring lipid-lowering teas.

[0060] Group Gynostemma pentaphyllum / g Danshen / g Hawthorn / g Bidens pilosa / g Cassia seed / g 1 5 5 5 5 10 2 5 10 10 10 10 3 5 15 15 15 10 4 10 5 10 15 10 5 10 10 15 5 10 6 10 15 5 10 10 7 15 5 15 10 10 8 15 10 5 15 10 9 15 15 10 5 10 total 90 90 90 90 90

[0061] Table 1

[0062] The results are shown in Table 2. Lipid-lowering Tea 8 scored the highest, therefore, it was selected as the formula for subsequent animal experiments. The formula consisted of Cassia Seed (10g), Gynostemma pentaphyllum (15g), Salvia miltiorrhiza (10g), Crataegus pinnatifida (5g), and Bidens pilosa (15g).

[0063]

[0064] Table 2

[0065] The maximum human intake was determined by querying the database, and the dosage for mice was obtained through the animal-to-human dosage conversion relationship. The dosage for each mouse was then determined based on its weight. The total dose required for gavage was calculated based on the final formula. The herbal decoction was then prepared, filtered, freeze-dried, and ground into a fine powder for later use.

[0066] 2.4 Animal Experiment Grouping and Drug Administration

[0067] Forty Kunming mice were randomly divided into four groups of ten each: a normal control group, a hyperlipidemia model group, a compound lipid-lowering tea group, and a simvastatin group. The normal control group was fed a diet rich in Vita-Like feed, while the other three groups were fed a high-fat diet, with each mouse receiving 7g of feed per day. All mice had free access to food and water. After acclimatization, the normal control group continued to be fed a regular diet, while the hyperlipidemia model group, simvastatin group, and lipid-lowering tea group were fed a high-fat diet for nine weeks. At the end of the ninth week, blood was collected from the tail vein to obtain serum samples to determine the concentration of total cholesterol (TC) in the fasting serum of the mice. A fasting TC value greater than 7 mmol / L was considered a successful model of hyperlipidemia.

[0068] Depend on Figure 8 It was observed that the mice's weight gradually increased with prolonged feeding time. After two weeks of high-fat diet feeding, the weight of mice in the hyperlipidemia model group, simvastatin group, and lipid-lowering tea group gradually exceeded that of the normal control group. By week 10, the weight of mice fed the high-fat diet was significantly higher than that of the normal control group. After week 10, the normal control group and hyperlipidemia model group were administered distilled water by gavage, while the simvastatin group and lipid-lowering tea group were administered simvastatin and lipid-lowering tea by gavage, respectively. Figure 8 As can be seen, after week 10, the body weight of mice in the normal control and hyperlipidemia model groups remained relatively stable, while the body weight of mice in the simvastatin and lipid-lowering tea groups showed a significant decreasing trend. At week 13, the body weight of mice in both groups was significantly lower than that of the normal control group. These results indicate that intervention with simvastatin and lipid-lowering tea can significantly reduce the body weight of mice, demonstrating a significant weight-loss effect.

[0069] 2.5 Effects of lipid-lowering tea on various organ indices in mice

[0070] Under normal growth conditions, the ratio of each organ to body weight is relatively constant. When animals become ill, the weight of damaged organs can change, thus altering the organ index. An increased organ index indicates organ congestion, edema, or hyperplasia and hypertrophy; a decreased organ index indicates organ atrophy and other degenerative changes. Compared to the normal control group, the fat index of the hyperlipidemia model group was significantly increased (p < 0.05), indicating a significant increase in body fat content in the hyperlipidemia model mice. A high-fat diet leads to the accumulation of adipose tissue, resulting in increased body weight. Compared to the hyperlipidemia model group, the fat index of the simvastatin group was significantly decreased (p < 0.05), indicating that drug intervention significantly reduced fat accumulation in mice, effectively breaking down adipose tissue and inhibiting fat accumulation caused by a high-lipidemia diet. Compared to the hyperlipidemia model group, the fat index of the lipid-lowering tea group was significantly reduced (p < 0.01), indicating that the lipid-lowering tea has a highly significant inhibitory effect on fat accumulation in mice (see...). Figure 9 A) Its effect is better than that of simvastatin, indicating that the compound lipid-lowering tea can effectively accelerate the decomposition of fat in high-fat mice, reduce fat accumulation, and demonstrate a good lipid-lowering effect.

[0071] Compared with the normal control group, the renal index of the hyperlipidemia model group was significantly lower (p < 0.05), indicating that a high-fat diet has an inhibitory effect on kidney growth and development, and may cause kidney tissue atrophy, resulting in adverse effects. Compared with the hyperlipidemia model group, the renal index of both the simvastatin group and the lipid-lowering tea group was significantly higher (p < 0.05), consistent with the renal index of the normal control group. Figure 9 B) indicates that simvastatin and lipid-lowering tea can effectively improve kidney atrophy caused by a high-fat diet.

[0072] Compared with the normal control group, the liver index in the hyperlipidemia model group was lower, but the difference was not statistically significant (p>0.05), indicating that a high-fat diet is not conducive to liver growth and development, but did not cause a significant difference. Compared with the hyperlipidemia model group, the liver index in the simvastatin group and the lipid-lowering tea group was higher, but the difference was not statistically significant (p>0.05), indicating that the drug intervention did not cause liver damage. Figure 9 C. Compared with the normal control group, the spleen, heart, testis, and epididymis indices of mice in the hyperlipidemia model group showed no significant changes (p>0.05), indicating that a high-fat diet had no adverse effects on the growth and development of these tissues and organs, and did not cause tissue swelling, congestion, or atrophy. Compared with the hyperlipidemia model group, the indices of the above organs in mice in the simvastatin group and the lipid-lowering tea group also showed no significant changes (p>0.05), indicating that simvastatin and lipid-lowering tea also had no adverse effects on the growth and development of these tissues and organs. Figure 9 DF.

[0073] 2.6 Effects of lipid-lowering tea on four lipid parameters in mouse serum

[0074] Mice in each group were fasted for 12 hours before the test, but allowed free access to water, to minimize the interference of food on the blood lipid test results. Blood was collected using the orbital venous plexus sampling method. During blood collection, the mice were held and fixed in place, and a capillary glass tube was inserted into the orbital venous plexus to collect blood, generally 0.2–0.5 mL. The collected blood sample was placed in a centrifuge tube and centrifuged at 3000–5000 rpm for 10–15 min. The serum was separated and carefully aspirated into a clean, dry centrifuge tube. Blood lipid indicators in the serum were then detected using a UV spectrophotometer. Following the instructions for detecting total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C), the serum sample was accurately added to the corresponding reagents, and the blood lipid indicator values ​​were read and recorded under a spectrophotometer for analysis.

[0075] After successful modeling, mice were administered the drug via gavage for 3 weeks. Serum lipid levels were then measured in each group of mice. Compared to the normal control group, the hyperlipidemia model group showed slightly elevated TC and LDL levels, but without significant differences (p > 0.05), while HDL levels were significantly decreased and TG levels were significantly increased (p < 0.05). This indicates that a high-fat diet induces abnormal lipid levels in mice, primarily by reducing HDL and increasing TG levels. Compared to the hyperlipidemia model group, the simvastatin group showed significantly decreased TC, LDL, and TG levels, while HDL levels were significantly increased (p < 0.05). This demonstrates that simvastatin has a significant lipid-lowering effect, mainly by reducing TC, LDL, and TG levels while simultaneously increasing HDL levels. Figure 10 As can be seen, compared with the hyperlipidemia model group, the TC and LDL levels in the lipid-lowering tea group mice were significantly reduced (p<0.05), the HDL level was significantly increased (p<0.05), and the TG level did not change significantly (p>0.05). This indicates that the lipid-lowering tea mainly regulates blood lipid components by reducing the synthesis of TC and LDL and increasing HDL levels. The lack of significant change in TG level may be due to the significant increase in HDL level.

[0076] 2.7 Oil Red O staining to detect lipid droplet formation in mouse liver tissue

[0077] Oil Red O dye exhibits excellent lipolysis and adsorption properties on tissues and organs with fatty degeneration and lipid deposition. It is superior to Sudan III dye due to its ease of use and bright color. After week 13, mice in each group were sacrificed by cervical dislocation, and liver tissue was harvested. The liver tissue was first fixed in 40 g / L paraformaldehyde solution for 4 hours, then transferred to 200 g / L sucrose solution-PB solution for dehydration (sucrose solution was changed multiple times). After 24 hours, the solution was changed to 300 g / L sucrose-PB solution, and after another 24 hours, the tissue was embedded in a embedding medium and finally frozen at -80°C for later use. Sections were prepared in a -20°C cryostat, spread, and air-dried at room temperature for 20 minutes before Oil Red O staining. Staining was performed with Oil Red O solution, differentiation with 60% isopropanol, counterstaining with hematoxylin, rinsing with running water, and mounting with glycerol gelatin or aqueous mounting medium. Changes in lipid droplets in the liver were observed and recorded using Oil Red staining. Specific staining steps are as follows: Figure 11 Oil Red O dye stains lipid droplets bright red and cell nuclei blue due to its lipolytic and adsorption properties.

[0078] When fatty degeneration occurs in liver tissue, intracellular fat aggregates into lipid bodies, which then fuse into lipid droplets, existing as vacuoles of varying sizes with some nuclei offset to one side of the cell. Figure 12The results showed that in the normal control group (A), uniform red granules appeared in hepatocytes, with small fat volume and light Oil Red O staining, indicating less and more uniform fat accumulation in the liver. In the hyperlipidemia model group (B), significant vacuolation was observed in the liver tissue. Compared with the normal control group, the intracellular lipid droplet volume was significantly larger and unevenly distributed within the cells, with deeper Oil Red O staining, indicating that the number and volume of lipid droplets in the liver cells of the hyperlipidemia model group mice were increased, resulting in greater fat accumulation. Red lipid droplets were also visible in the hepatocytes of the simvastatin group (C), but their number and volume were smaller than those in the hyperlipidemia model group. The traditional Chinese medicine lipid-lowering drug group (D) also showed a small number of fine lipid droplets in the hepatocytes, significantly reduced compared to the hyperlipidemia model group, with uniform and lighter Oil Red O staining. These results indicate that simvastatin and lipid-lowering drugs can significantly inhibit the formation of fat in liver cells, thereby reducing the number and volume of lipid droplets and inhibiting fat accumulation in liver cells, thus exerting a lipid-lowering effect.

[0079] 2.8 HE staining was used to observe the histopathological characteristics of liver tissue in each group of mice.

[0080] Liver tissue was collected from each group of mice according to... Figure 13 The steps involved preparing paraffin sections and staining them with hematoxylin and eosin (HE). The sections were then observed and compared under a microscope: low magnification revealed the structure, arrangement, and shape of the central vein in the liver lobules; high magnification revealed the nuclei, cytoplasm, number, size, shape, and surrounding structures of the hepatocytes.

[0081] After HE staining, the results were as follows: Figure 14 As shown, in the normal control group, the liver cells in the liver tissue had purplish-red cytoplasm and blue nuclei with one to several large, round, and prominent nucleoli. The nuclear membrane was clear, and the intranuclear structure was relatively loose. The liver cells were arranged neatly and regularly, radiating outwards from the central vein. In contrast, the liver tissue of the hyperlipidemia model group showed obvious vacuoles, appearing as round, well-defined, colorless areas, with a higher concentration near the central vein. The cytoplasm of the liver cells was darker, and the nuclei were smaller and darker. In contrast, no vacuoles were observed in the liver tissue of mice in the simvastatin group and the lipid-lowering tea group. The cells were tightly packed, with darker cytoplasm staining, smaller nuclei, and darker nuclear staining. This indicates that a high-lipidemia diet increases intracellular fat in liver cells, leading to excessive fat accumulation. Excessive fat accumulation affects normal liver function, resulting in vacuolar formation. Simvastatin and the lipid-lowering tea can reduce lipid droplet formation in liver cells and improve the vacuolar phenomenon in liver tissue, demonstrating a good lipid-lowering effect.

[0082] In summary, this invention, through network pharmacology analysis and animal experiments, explored the lipid-lowering effects and mechanisms of lipid-lowering tea from multiple perspectives. The study confirmed that the main lipid-lowering active ingredients in the tea are tanshinone, flavonoids, ursolic acid, and anthraquinones. These active ingredients exert their lipid-lowering effects by targeting various signaling pathways, including the PI3K-AKT signaling pathway, cancer pathogenesis pathways, estrogen signaling pathways, endocrine resistance, and tyrosine kinase inhibitors. Flavonoids can bind to the core target PPAGR, while anthraquinones and ursolic acid bind to the core targets HSP90AA and ESR1, respectively, regulating the activity of these target proteins and affecting lipid synthesis or metabolism, thereby inhibiting adipose tissue accumulation and reducing the lipid index. Simultaneously, it can also reduce the synthesis of low-density lipoprotein and triglycerides in serum, promote the synthesis of high-density lipoprotein, inhibit the formation of lipid droplets in liver cells, and reduce lipid growth, thus exerting its lipid-lowering effects through multiple aspects and pathways.

[0083] The above descriptions are merely embodiments of the present invention, and common knowledge regarding specific structures and characteristics is not elaborated upon here. It should be noted that those skilled in the art can make various modifications and improvements without departing from the structure of the present invention, and these should also be considered within the scope of protection of the present invention. These modifications and improvements will not affect the effectiveness of the present invention or the practicality of the patent. The scope of protection claimed in this application should be determined by the content of its claims, and the specific embodiments described in the specification can be used to interpret the content of the claims.

Claims

1. A compound lipid-lowering tea, characterized in that: The Chinese medicinal ingredients consist of 10g of Cassia Seed, 5-15g of Gynostemma pentaphyllum, 5-15g of Salvia miltiorrhiza, 5-15g of Crataegus pinnatifida, and 5-15g of Bidens pilosa.

2. The compound lipid-lowering tea according to claim 1, characterized in that: Cassia seed 10g, Gynostemma pentaphyllum 5g, Salvia miltiorrhiza 5g, Crataegus pinnatifida 5g, Bidens pilosa 5g.

3. The compound lipid-lowering tea according to claim 1, characterized in that: Cassia seed 10g, Gynostemma pentaphyllum 5g, Salvia miltiorrhiza 10g, Crataegus pinnatifida 10g, Bidens pilosa 10g.

4. The compound lipid-lowering tea according to claim 1, characterized in that: Cassia seed 10g, Gynostemma pentaphyllum 5g, Salvia miltiorrhiza 15g, Crataegus pinnatifida 15g, Bidens pilosa 15g.

5. The compound lipid-lowering tea according to claim 1, characterized in that: Cassia seed 10g, Gynostemma pentaphyllum 10g, Salvia miltiorrhiza 5g, Crataegus pinnatifida 10g, Bidens pilosa 15g.

6. The compound lipid-lowering tea according to claim 1, characterized in that: Cassia seed 10g, Gynostemma pentaphyllum 15g, Salvia miltiorrhiza 15g, Crataegus pinnatifida 10g, Bidens pilosa 5g.

7. The compound lipid-lowering tea according to claim 1, characterized in that: Cassia seed 10g, Gynostemma pentaphyllum 10g, Salvia miltiorrhiza 15g, Crataegus pinnatifida 5g, Bidens pilosa 10g.

8. The compound lipid-lowering tea according to claim 1, characterized in that: Cassia seed 10g, Gynostemma pentaphyllum 15g, Salvia miltiorrhiza 10g, Crataegus pinnatifida 5g, Bidens pilosa 15g.

9. A method for preparing a compound lipid-lowering tea according to any one of claims 1-8, characterized in that: Includes the following steps: S1. Pretreatment: Clean and dry each Chinese herb, then grind it into powder using a Chinese herb grinder. S2. Weighing: Weigh the required weights of cassia seeds, gynostemma pentaphyllum, salvia miltiorrhiza, hawthorn, and beggar-ticks using a weighing device, and soak the mixed powder in an appropriate amount of water for a period of time. S3. Boil the soaking liquid in S2 over high heat for a period of time, then boil the soaking liquid over low heat for a period of time. S4. Filter the decoction in S3 using gauze and collect the filtrate. S5. Add an appropriate amount of water to the filtered dregs and boil over high heat for a period of time, then boil over low heat for a period of time. S6. Filter the decoction in S5 using gauze and combine the filtrates; S7. The filtrate is concentrated in a water bath to obtain the compound lipid-lowering tea.

10. A method for preparing a compound lipid-lowering tea according to claim 9, characterized in that: In step S2, the mixed powder is soaked in 1 L of purified water for half an hour; in step S2, the soaking liquid is first boiled over high heat for 30 minutes, and then boiled over low heat for 1 hour; in step S5, the filtered residue is added to 0.5 L of purified water and boiled over high heat for 30 minutes, and then boiled over low heat for 1 hour; in step S7, the filtrate is concentrated to 100 mL in a 100℃ water bath to obtain the compound lipid-lowering tea.

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