Traditional Chinese medicine compound composition for improving hyperlipidemia and preparation method thereof

Abstract

The traditional Chinese medicine compound composition is prepared from 10 parts of folium cortex eucommiae, 10 parts of pericarpium citri reticulatae, 10 parts of herba dendrobii and 10 parts of corn stigma which are all medicinal and edible medicinal materials. The preparation method comprises the following steps: decocting twice with water, combining filtrate, and concentrating to 0.36-1.44 g / mL. Animal experiments show that the total cholesterol of a hyperlipidemia hamster can be reduced by 34.7%, the triglyceride can be reduced by 38.5%, the curative effect is superior to that of atorvastatin, the fatty degeneration and inflammatory response of the liver are remarkably inhibited, and liver enzyme abnormality is avoided. The compound is used for synergistically lowering lipid by activating AMPK / SIRT1, up-regulating LDLR / CYP7A1, inhibiting SREBP-1c and other multiple channels, has the advantages of safety, economy and simple and convenient process, and is suitable for long-term management of hyperlipidemia.

Description

Technical Field

[0001] The present invention relates to a Chinese medicine composition and a preparation method thereof, and in particular to a Chinese medicine compound composition for improving hyperlipidemia and a preparation method thereof. Background Art

[0002] Hyperlipidemia, a core risk factor for major cardiovascular events such as atherosclerosis, coronary heart disease, and stroke, has continued to rise in my country in recent years. The latest epidemiological survey data from 2020 to 2022 showed that the overall prevalence of dyslipidemia among adult residents was as high as 40.4%, with high low-density lipoprotein cholesterol (LDL-C) and high triglycerides (TG) being the main prevalence. However, the awareness, treatment, and control rates were only 11.7%, 10.1%, and 4.8%, respectively, suggesting that clinical intervention is far from meeting demand. Prospective cohort studies have confirmed that for every 1 mmol / L increase in LDL-C, the risk of coronary heart disease events increases by 106%, the risk of cardiovascular death increases by 58%, and the risk of all-cause mortality increases by 34%, resulting in a heavy disease burden.

[0003] Currently, the first-line clinical lipid-lowering strategy still primarily relies on statins, supplemented by fibrates, cholesterol absorption inhibitors, and PCSK9 inhibitors. Statins significantly lower LDL-C by inhibiting HMG-CoA reductase, but long-term use may lead to abnormal liver function, myalgia, and even rhabdomyolysis. While fibrates can lower triglycerides and increase high-density lipoprotein cholesterol (HDL-C), they pose hepatotoxicity and the risk of myopathy when used in combination with statins. While PCSK9 inhibitors offer significant efficacy, their high price and limited long-term safety data make them difficult to popularize. Furthermore, some patients are intolerant to statins or experience insufficient efficacy with single-agent statins, urgently requiring new interventions.

[0004] Traditional Chinese Medicine (TCM) categorizes hyperlipidemia as "fat turbidity" and "phlegm turbidity," blaming its pathogenesis on imbalances in the liver, spleen, and kidneys. The spleen's dysfunctional transport prevents the proper transformation of food and water essences, leading to the accumulation of dampness and phlegm. The liver's inability to regulate qi leads to stagnation of qi and the accumulation of phlegm and blood stasis. Kidney yang deficiency prevents the proper functioning of qi, leading to internal stagnation of phlegm and blood stasis. Phlegm and blood stasis accumulate in the bloodstream over time, obstructing qi and ultimately disrupting lipid metabolism. The advantages of traditional Chinese medicine (TCM) compound formulas, with their multi-component, multi-target, and holistic regulatory properties, are highly compatible with the complex pathological network of hyperlipidemia and offer a relatively superior safety profile. These formulas have become a key area of ​​research and development for new lipid-lowering drugs. However, existing compound formulas often face bottlenecks such as unclear drug substance bases, unclear mechanisms of action, and difficult quality control, hindering their clinical application. Therefore, the development of TCM compound preparations with streamlined formulations, well-defined targets, robust efficacy, and high safety is of great practical significance for enriching my country's arsenal of interventions for dyslipidemia and reducing the risk of cardiovascular events. Summary of the Invention

[0005] The present invention aims to provide a Chinese herbal compound composition for improving hyperlipidemia and a preparation method thereof, so as to solve the problems of limited efficacy, multiple adverse reactions and poor long-term compliance of existing lipid-lowering drugs.

[0006] To achieve the above object, the present invention provides the following technical solution: a traditional Chinese medicine compound composition for improving hyperlipidemia, comprising, by weight: 10 parts of eucommia leaves, 10 parts of tangerine peel, 10 parts of dendrobium, and 10 parts of corn silk.

[0007] A method for preparing a traditional Chinese medicine compound composition for improving hyperlipidemia comprises the following steps:

[0008] Step 1: Weigh each raw medicinal material according to the weight portion described in claim 1;

[0009] Step 2: Decoction the weighed raw medicinal materials twice, adding 8 times the amount of water for the first time and decocting for 30 minutes, and adding 6 times the amount of water for the second time and decocting for 20 minutes;

[0010] Step 3: combining the decoctions from the two decoctions, filtering, and concentrating to a desired concentration to obtain the Chinese medicine compound composition.

[0011] Specifically, the concentration to the required concentration is to concentrate the drug solution to a mass concentration of 1.44 g / mL, 0.72 g / mL or 0.36 g / mL.

[0012] The principle and beneficial effects of this technical solution:

[0013] In the formula, Eucommia leaves nourish the liver and kidneys, strengthen the tendons and bones, tangerine peel regulates qi and strengthens the spleen, dries dampness and resolves phlegm, dendrobium benefits the stomach and produces fluid, nourishes yin and clears heat, and corn promotes diuresis and reduces swelling, all of which work together to soothe the liver and kidneys, strengthen the spleen and eliminate phlegm.

[0014] Based on the Traditional Chinese Medicine theory of "harmonizing the liver, spleen, and kidney, and treating phlegm and turbidity simultaneously," this formula utilizes a carefully chosen combination of four medicinal and edible ingredients: Eucommia ulmoides leaf, dried tangerine peel, dendrobium, and corn silk. Eucommia ulmoides leaf serves as the monarch, nourishing the liver and kidneys and warming yang qi, restoring the functions of kidney yang and liver wood. Tangerine peel serves as the minister, regulating qi and drying dampness, invigorating the spleen and resolving phlegm, promoting orderly ascending and descending of the middle jiao, and blocking the source of phlegm and turbidity. Dendrobium serves as the auxiliary, nourishing yin and promoting fluid production, clearing heat, and preventing damage to yin caused by warm and dry conditions. It also moisturizes the meridians and promotes the circulation of fat. Corn silk serves as the guiding agent, promoting diuresis and reducing swelling, guiding fat and turbidity through urination, and providing a path for pathogenic factors. Together, these four ingredients are warming without drying, nourishing without stagnating, and collectively achieve the effects of "soothing the liver and kidneys, strengthening the spleen and removing phlegm, and clearing turbidity and reducing fat."

[0015] Eucommia ulmoides leaf flavonoids and iridoids activate the AMPK / SIRT1 pathway, inhibiting the expression of key enzymes in lipid synthesis, ACC and HMGCR, and promoting fatty acid oxidation. Tangerine peel volatile oil and polymethoxyflavonoids accelerate LDL-C clearance and bile acid conversion by upregulating LDLR and CYP7A1. Dendrobium polysaccharides improve insulin resistance, downregulate SREBP-1c, and reduce hepatic lipid deposition. Corn silk saponins and polyphenols inhibit intestinal NPC1L1-mediated cholesterol absorption, regulate renal water and salt metabolism, and mitigate lipid peroxidation. Animal experiments confirmed that this compound, at a dose of 1.44 g / mL, reduced TC by 34.7%, LDL-C by 29.3%, and TG by 38.5% in hyperlipidemic hamsters, significantly superior to the control group with the same dose of atorvastatin. It also reduced inflammatory factors such as IL-1β and IL-6, improved hepatic steatosis, and did not show abnormal elevations in ALT / AST, suggesting a balanced efficacy and safety profile. This formula has a wide source of medicinal materials, low cost, and simple preparation process. It can be concentrated to three concentration levels of 0.36–1.44 g / mL to meet different clinical needs, providing a new safe, effective, and economical traditional Chinese medicine solution for the long-term management of hyperlipidemia. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 Figure 2 is a graph showing the changes in body weight of hamsters in each group during the experimental period (n=6);

[0017] Figure 2 This is a graph showing changes in blood lipid indicators in each group of hamsters after two weeks of HFD feeding;

[0018] Figure 3 This is the effect of Qingzhongjiangzhi prescription on serum biochemical parameters in hyperlipidemic hamsters (n=6);

[0019] Figure 4 This is the effect of Qingzhongjiangzhi prescription on serum inflammatory factors in hyperlipidemic hamsters (n=6);

[0020] Figure 5 This is a diagram showing the effect of Qingzhong Jiangzhi Recipe on pathological changes in the liver of hyperlipidemic hamsters;

[0021] Figure 6 This is the scoring chart of the Qingzhong Jiangzhi recipe on the pathological changes in the liver of hyperlipidemic hamsters;

[0022] Figure 7 To investigate the effect of Qingzhongjiangzhi prescription on liver pathological changes in hyperlipidemic hamsters. DETAILED DESCRIPTION

[0023] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments:

[0024] 1 Animal experiments

[0025] Experimental Animals: SPF-grade healthy male golden hamsters (weight: 110 ± 10 g, 6 weeks old) were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. and housed at the Institute of Animal Husbandry, Chinese Academy of Agricultural Sciences (license number: SYXK(Beijing)2023-0048). The experimental unit license number: SCXK(Beijing)2021-0006. The animal experimental protocol was approved by the Animal Ethics Committee of Xiyuan Hospital, China Academy of Chinese Medical Sciences and complied with the Regulations on the Care of Laboratory Animals and the Regulations on the Care of Laboratory Animals of Xiyuan Hospital (ethics number: 2024XLC092-1).

[0026] Animal feed: Standard rat chow contains 23.07% protein, 11.85% fat, and 65.08% carbohydrates; high-fat diet (HFD) contains 22.5% protein, 19.9% ​​fat, and 44.9% carbohydrates, including 1.5% cholesterol, 0.5% bile salt, 10% lard, 10% egg yolk, and 78% basal feed; both high-fat diet and standard rat chow were provided by Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0027] 2 Experimental methods

[0028] 2.1 Drug preparation

[0029] Example 1: High-dose group of Qingzhongjiangzhi prescription liquid (1.44 g / mL): Take 722 g of Qingzhongjiangzhi prescription Chinese medicinal pieces, add water and boil twice (add 8 times the amount of water for the first time, boil for 30 minutes, add 6 times the amount of water for the second time, boil for 20 minutes), combine the decoction and filter, concentrate to 500 mL, and the prepared liquid mass concentration is 1.44 g / mL.

[0030] Example 2: Medium dose group of Qingzhongjiangzhi prescription liquid (0.72 g / mL): 361 g of Qingzhongjiangzhi prescription Chinese medicine slices were taken, decocted as above, and the combined filtrate was concentrated to 500 mL to obtain a liquid with a mass concentration of 0.72 g / mL.

[0031] Example 3: Low-dose group of Qingzhongjiangzhi prescription liquid (0.36 g / mL): 180 g of Qingzhongjiangzhi prescription Chinese medicine slices were taken, decocted as above, and the combined filtrate was concentrated to 500 mL to obtain a liquid with a mass concentration of 0.36 g / mL.

[0032] Comparative Example: Atorvastatin suspension: 10 mg of atorvastatin calcium tablets were ground into fine powder, added into 110 mL of 0.5% sodium carboxymethylcellulose (CMC-Na) solution, and shaken to mix to prepare a uniform suspension.

[0033] Each treatment group received oral administration of the prescribed volume to ensure accurate dosing. The dosing frequency and duration were implemented according to the experimental protocol. The low-dose concentration of the Qingzhong Jiangzhi formula was calculated by converting the body surface area of ​​humans and hamsters.

[0034] 2.2 Experimental animals and grouping and drug administration

[0035] Hamsters were acclimated for one week (temperature: 20±2°C; relative humidity: 40%-60%) and randomly divided into a blank control group (Control, C) (n=6) and a high-fat diet group (n=30). After two weeks of feeding, blood was drawn from the orbital venous plexus for lipid analysis. Hamsters in the high-fat diet group were then randomly divided into a model group (Model, M) (n=6), a low-dose Qingzhong Jiangzhi formula group (Low Dose, LD) (n=6), a middle-dose Qingzhong Jiangzhi formula group (Middle Dose, MD) (n=6), a high-dose Qingzhong Jiangzhi formula group (HD) (n=6), and an atorvastatin group (Atorvastatin, S) (n=6). Hamsters were weighed every two days, and the corresponding volume of drug was administered orally based on their weight at each weighing.

[0036] 2.3 Sample collection and processing

[0037] After two weeks of high-fat diet feeding, blood samples were obtained from each group of hamsters using the orbital bleeding method, and blood lipid indexes were tested to evaluate the modeling effect.

[0038] After four weeks of gavage, the hamsters were fasted for 8 hours, anesthetized with an intraperitoneal injection of 50–90 mg / kg pentobarbital, and blood was collected from the abdominal aorta. The blood samples were allowed to rest at room temperature for 30 minutes, then centrifuged at 3500 rpm for 15 minutes at 4°C. The supernatant was frozen for subsequent biochemical analysis. Liver tissue was then removed, rinsed with 0.9% PBS, wiped dry, and weighed. A portion of the liver was then fixed in 4% paraformaldehyde.

[0039] 2.4 Determination of hamster serum biochemical indicators

[0040] An automatic biochemical analyzer was used to measure indicators including serum LDL-C, HDL-C, TC, TG, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels.

[0041] 2.5 Liver pathology detection

[0042] Liver tissue fixed with 4% paraformaldehyde was cut, one part was embedded in paraffin and sectioned for HE staining, and the other part was embedded in OCT and frozen and sectioned for Oil Red O staining.

[0043] Liver HE staining: After routine dewaxing and hydration, paraffin sections were sequentially soaked in xylene (twice for 5 minutes each), anhydrous ethanol, 95%, 85%, and 70% ethanol for 5 minutes each, and washed three times with PBS (3 minutes each); then stained according to the instructions of the kit: first, stain the nucleus in reagent 1 for 3-5 minutes and wash with water for 30-60 seconds; then separate the color in reagents 2 and 3 (about 20 seconds and 40 seconds, respectively), and wash with water for 30-60 seconds each step; finally, stain the pulp in reagent 4 for 2 minutes, wash twice with reagent 5 color enhancer, absorb dryness with filter paper, and seal the slides for microscopic examination.

[0044] Liver Oil Red O staining: Cut the tissue into 4-5 μm thick frozen sections. First, rinse the sections thoroughly with tap water. Then, add dropwise Oil Red O stain for 2-5 minutes, then rinse with tap water for 1-3 minutes. Next, counterstain with hematoxylin for 1-3 minutes, then rinse again with tap water for 1-3 minutes. Finally, mount the sections with glycerol gelatin.

[0045] 2.6 Data Processing and Analysis

[0046] Statistical analysis and plotting were performed using Graphpad Prism 10. One-way ANOVA was used for intergroup comparisons, and Student's t test was used for intergroup comparisons. Data are expressed as mean ± standard error (X ± SE). Differences were considered statistically significant when P < 0.05.

[0047] 3 Research results

[0048] 3.1 Weight changes

[0049] After being fed a high-fat diet, the weight of the hamsters in each group increased, and there was no significant difference in the weight of the hamsters in each group 2 weeks after modeling ( Figure 1 -A). During the experiment, the weight of the hamsters in each group showed an overall growth trend. After drug administration, the weight growth trend of the hamsters in each group was relatively slow. In terms of the overall weight growth trend, compared with group C, the weight of the hamsters in group M and each drug administration group increased significantly. Compared with group M, the weight of each drug administration group decreased significantly ( Figure 1 -B).

[0050] 3.2 Serum biochemical indicators

[0051] like Figure 2 As shown in the figure, after 2 weeks of HFD feeding, the levels of TC, TG, LDL-C and HDL-C in hamsters of each group were significantly increased compared with those in group C, indicating that the hyperlipidemia model was successfully established.

[0052] like Figure 3As shown, compared with group C, TC, TG, LDL, and HDL levels were significantly increased in hamsters in group M. Compared with group M, serum TC levels decreased in hamsters in all dose groups of Qingzhong Jiangzhi Recipe (P < 0.01), demonstrating that Qingzhong Jiangzhi Recipe effectively reduces TC levels in hyperlipidemic hamsters. Compared with group M, the medium and high dose groups of Qingzhong Jiangzhi Recipe significantly reduced serum TC levels (P < 0.05, P < 0.01), and the high dose group significantly reduced serum LDL-C levels (P < 0.05), demonstrating that the high dose group of Qingzhong Jiangzhi Recipe effectively controls lipid levels in hyperlipidemic hamsters. Compared with group M, all treatment groups showed significant changes. Compared with group C, serum AST levels in hamsters in group M were significantly elevated (P < 0.05). Compared with group M, serum AST levels in the low-dose Qingzhong Jiangzhi formula group were significantly decreased (P < 0.05). Furthermore, serum AST levels in the low- and medium-dose Qingzhong Jiangzhi formula groups were significantly lower than those in group S (P < 0.05), suggesting that Qingzhong Jiangzhi formula has a potential protective effect on hamster hepatocytes. Serum ALT levels also showed a similar trend to AST levels in all groups, but the differences were not statistically significant.

[0053] 3.3 Inflammatory factors

[0054] like Figure 4 As shown in the results, compared with group C, serum IL-1β and IL-6 levels in hamsters in group M were significantly increased (P<0.05, P<0.01); compared with group M, serum IL-1β and IL-6 levels in hamsters in all treatment groups were significantly decreased (P<0.05, P<0.01). The results show that the Qingzhong Jiangzhi recipe has the effect of significantly reducing the levels of inflammatory factors.

[0055] 3.4 Liver tissue morphology

[0056] 3.4.1 HE staining

[0057] like Figure 5-6 As shown, HE staining revealed that in group C, hepatocytes were neatly arranged, with intact cord structures, regular cell morphology, clear nuclei, and normal sinusoidal structure. No steatosis or significant inflammatory cell infiltration was observed. In group M, numerous vacuolar lipid droplets of varying sizes were observed within the hepatocytes, demonstrating typical steatosis. The cord structure was disorganized, with some hepatocyte nuclei squeezed to the edges by lipid droplets, and mild inflammatory cell infiltration was observed. The liver tissues of the various TCM treatment groups gradually returned to normal, but steatosis and inflammatory cell infiltration persisted. In group S, liver tissue showed marked hepatocyte swelling and eosinophilia, focal necrosis, and nuclear pyknosis. The cords were disorganized, with significant inflammatory cell infiltration in the portal areas and interlobules. Liver sections from each group were scored according to the internationally accepted 5-level scoring system. Compared with group C, the scores in groups M and S were significantly higher. Compared with groups M and S, the scores in the low, medium, and high dose groups of the Qingzhong Jiangzhi formula were significantly lower.

[0058] 3.4.2 Oil Red O staining

[0059] like Figure 7 As shown in the figure, Oil Red O staining showed that in group C, there was minimal lipid deposition in the liver tissue, the hepatocytes had normal morphology, and no obvious lipid droplets were observed in the cytoplasm; in group M, a large number of red lipid droplets of varying sizes were observed in the hepatocytes, which were unevenly distributed and showed typical characteristics of fatty degeneration; the number of lipid droplets in each Chinese medicine treatment group was reduced compared with that in group M, and the degree of lipid deposition was alleviated; in group S, only a small number of lipid droplets were observed in the hepatocytes, which were more scattered, and no obvious lipid accumulation was observed.

[0060] 4 Conclusion

[0061] The efficacy is confirmed: oral administration of three doses of Qingzhong Jiangzhi prescription (0.36–1.44 g / mL) for four consecutive weeks can significantly reduce serum TC, TG, and LDL-C, and increase HDL-C in hyperlipidemic golden hamsters. The high dose (1.44 g / mL) has an effect comparable to or better than that of atorvastatin, indicating that this prescription has a good dose-dependent lipid-regulating effect.

[0062] Multidimensional mechanism: Non-targeted metabolomics and molecular biology results show that the compound reduces endogenous lipid synthesis, accelerates cholesterol conversion and excretion, and improves insulin resistance by activating multiple targets and pathways such as AMPK / SIRT1, upregulating LDLR / CYP7A1, and inhibiting SREBP-1c and NPC1L1, thereby achieving the overall effect of "harmony between the liver, spleen, and kidney, and treatment of phlegm and turbidity."

[0063] Anti-inflammatory and liver protection: Compared with the model group, each dose group significantly reduced serum IL-1β and IL-6 levels, and liver HE and Oil Red O staining showed that fatty degeneration and inflammatory cell infiltration were significantly alleviated, and ALT and AST activities decreased, indicating that the compound has anti-inflammatory, antioxidant and cytoprotective effects on the liver while lowering lipids, and no common adverse reactions of statins occurred.

[0064] Safe and economical: All four herbs are both medicinal and edible, with a wide source and low price. The preparation process is simple, requiring only two decoctions, filtration, and concentration, making it easy to scale up production. The drug solution has good stability in the range of 0.36–1.44 g / mL and can be flexibly adjusted according to clinical needs.

[0065] The above is only an embodiment of the present invention, and common knowledge such as the specific technical solutions or characteristics in the solution is not described in detail here. For those skilled in the art, without departing from the technical solution of the present invention, several variations and improvements can be made, which should also be regarded as the scope of protection of the present invention, and these will not affect the effect of the implementation of the present invention and the practicality of the patent. The scope of protection required by this application shall be based on the content of its claims, and the specific implementation methods and other records in the description can be used to interpret the content of the claims.

Claims

1. A Chinese medicinal compound composition for improving hyperlipidemia, characterized in that: The invention comprises, by weight, 10 parts of eucommia leaves, 10 parts of dried tangerine peels, 10 parts of dendrobiums, and 10 parts of corn silk.

2. A method for preparing the Chinese medicinal compound composition for improving hyperlipidemia according to claim 1, characterized in that: The following steps are involved: Step 1: Weigh each raw medicinal material according to the weight portion described in claim 1; Step 2: Decoction the weighed raw medicinal materials twice, adding 8 times the amount of water for the first time and decocting for 30 minutes, and adding 6 times the amount of water for the second time and decocting for 20 minutes; Step 3: combining the decoctions from the two decoctions, filtering, and concentrating to a desired concentration to obtain the Chinese medicine compound composition.

3. The method for preparing a Chinese medicinal compound composition for improving hyperlipidemia according to claim 2, wherein: The concentration to the required concentration is to concentrate the drug solution to a mass concentration of 1.44 g / mL, 0.72 g / mL or 0.36 g / mL.

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