A traditional Chinese medicine composition for treating kawasaki disease and a preparation method thereof

Abstract

The present application relates to the technical field of Chinese medicine preparation, and particularly relates to a traditional Chinese medicine composition for treating Kawasaki disease and a preparation method thereof.The traditional Chinese medicine composition is prepared from raw medicinal materials including the following components by weight: radix rehmanniae 8-30 parts, forsythia suspensa 2-10 parts, cortex moutan 2-10 parts, pseudo-ginseng 2-10 parts, giant knotweed 2-10 parts, radix scrophulariae 2-10 parts, long-jointed willow root 2-10 parts, radix clematidis 1-5 parts and radix puerariae 3-15 parts.The traditional Chinese medicine composition is a pure traditional Chinese medicine preparation, has the effects of clearing heat and cooling blood, promoting blood circulation and detoxification, and is suitable for treating Kawasaki disease (especially suitable for heat-toxin blood-stasis type).Experimental results show that the traditional Chinese medicine composition has a good therapeutic effect on Kawasaki disease.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, and in particular to a traditional Chinese medicine composition for treating Kawasaki disease and its preparation method. Background Technology

[0002] Kawasaki disease (KD), also known as mucocutaneous lymph node syndrome, is an acute febrile illness characterized primarily by systemic vasculitis. It mainly affects small and medium-sized blood vessels, and its typical clinical manifestations include persistent fever, diffuse hyperemia of the oral mucosa, red and cracked lips, strawberry tongue, erythema multiforme rash, early induration of the hands and feet followed by membranous desquamation, and acute non-suppurative cervical lymphadenopathy. The most serious harm of this disease lies in its cardiovascular complications, which can lead to coronary artery dilation, coronary artery stenosis, and even coronary aneurysm, making it the leading cause of acquired heart disease in children. Epidemiological studies show that although KD primarily affects children under 5 years old, it can also affect other age groups, with a significant gender difference; the incidence rate in males is approximately 1.5 times that in females. The incidence rate is lower in infants under 4 months of age, presumably related to the protective effect of maternal antibodies. The aforementioned epidemiological characteristics strongly suggest that genetic factors play a decisive role in the pathogenesis of Kawasaki disease, while environmental factors also participate as triggering conditions. Notably, the incidence of Kawasaki disease has decreased or remained stable due to reduced exposure to pathogens in the population, further supporting the important role of infectious factors in disease triggering.

[0003] Currently, the medical community has not reached a unified consensus on the exact cause and pathogenesis of Kawasaki disease. It is generally believed that infection, genetic susceptibility, and abnormal activation of the immune system are the three main factors in the development and progression of the disease. In modern medical treatment, the treatment of Kawasaki disease mainly relies on drug intervention, with the standard treatment regimen being intravenous immunoglobulin combined with oral aspirin. In addition, glucocorticoids, tumor necrosis factor-α antagonists, and plasma exchange are also used as adjunctive therapies. However, the above treatment regimens have many limitations: some children do not respond to immunoglobulin therapy, requiring adjustments to the treatment plan; long-term use of aspirin can cause various adverse reactions such as gastrointestinal reactions, liver damage, and Reye's syndrome; more importantly, even with standard treatment, some children still face the long-term risk of coronary artery damage, and their prognosis is not entirely optimistic, with the possibility of myocardial infarction.

[0004] In the field of Traditional Chinese Medicine (TCM), there is no direct corresponding disease name for Kawasaki disease in ancient TCM literature. Based on its clinical manifestations and transmission patterns, modern physicians mostly classify it under the category of "febrile diseases," considering it similar to rashes, epidemic rashes, febrile toxins, and yang-toxin rashes. Most physicians believe that the pathogenesis of Kawasaki disease involves the entry of warm-heat pathogens through the mouth and nose, accumulating in the lungs and stomach, and its transmission follows the principles of Wei Qi Ying Xue (defensive Qi, nutritive Qi, and blood), with blood stasis persisting throughout the course of the disease. Based on this understanding of the pathogenesis, current TCM clinical treatment mainly focuses on tonifying Qi and nourishing Yin, promoting blood circulation and removing blood stasis, and clearing heat and detoxifying. It often employs the theory of Wei Qi Ying Xue in febrile diseases for syndrome differentiation and treatment, or physicians create specific prescriptions based on their personal clinical experience. In addition, there are reports of using compound Danshen injection to regulate capillary tension, reduce platelet aggregation, and lower blood viscosity; using Shengmai liquid to alleviate the sequelae of ischemic heart disease; and using Shuanghuanglian for antibacterial, antiviral, and immune-regulating effects. However, existing TCM treatment options still have significant shortcomings. First, while syndrome differentiation and treatment embodies the advantages of individualized therapy, there is a lack of standardized compound preparations specifically for children. Clinical use primarily relies on decoctions with modifications based on syndrome differentiation, leading to poor patient compliance. Second, existing traditional Chinese medicine preparations or injections are not designed specifically for the core pathogenesis of Kawasaki disease and the physiological characteristics of children, resulting in imprecise clinical positioning and difficulty in meeting the treatment needs of different stages of Kawasaki disease. Therefore, developing a traditional Chinese medicine compound preparation that targets the etiology and pathogenesis of Kawasaki disease, has definite efficacy, an appropriate dosage form, good pediatric compliance, and a reasonable price has become an urgent technical problem to be solved in this field.

[0005] In view of this, the present invention is hereby proposed. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention provides a traditional Chinese medicine composition for treating Kawasaki disease and its preparation method.

[0007] Specifically, the technical solution of the present invention is as follows: In a first aspect, the present invention provides a traditional Chinese medicine composition for treating Kawasaki disease, the traditional Chinese medicine composition being prepared from raw materials comprising the following parts by weight: 8-30 parts of Rehmannia glutinosa, 2-10 parts of Forsythia suspensa, 2-10 parts of Paeonia suffruticosa, 2-10 parts of Paeonia lactiflora, 2-10 parts of Polygonum cuspidatum, 2-10 parts of Scutellaria baicalensis, 2-10 parts of Cynanchum paniculatum, 1-5 parts of Cimicifuga foetida, and 3-15 parts of Pueraria lobata.

[0008] Preferably, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 8-15 parts of Rehmannia glutinosa, 2-5 parts of Forsythia suspensa, 2-5 parts of Paeonia suffruticosa, 2-5 parts of Paeonia lactiflora, 2-5 parts of Polygonum cuspidatum, 2-5 parts of Scutellaria baicalensis, 2-5 parts of Cynanchum paniculatum, 1-2.5 parts of Cimicifuga foetida, and 3-7.5 parts of Pueraria lobata.

[0009] Preferably, the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 15 parts Rehmannia glutinosa, 5 parts Forsythia suspensa, 5 parts Paeonia suffruticosa, 5 parts Paeonia lactiflora, 5 parts Polygonum cuspidatum, 5 parts Scutellaria baicalensis, 5 parts Cynanchum paniculatum, 2.5 parts Cimicifuga foetida, and 7.5 parts Pueraria lobata.

[0010] Secondly, the present invention provides a traditional Chinese medicine preparation, which is made solely from the aforementioned traditional Chinese medicine composition, or made from the traditional Chinese medicine composition with a pharmaceutically acceptable carrier and / or excipient. Preferably, the dosage form of the traditional Chinese medicine preparation is tablets, pills, capsules, granules, powders, ointments, oral liquids, or mixtures.

[0011] Thirdly, the present invention provides a method for preparing the aforementioned traditional Chinese medicine preparation, comprising the following steps: S1. Weigh each raw material according to its weight parts; S2. Place the raw material in the extraction solvent, separate, and obtain the extract; S3. Combine the extract with a carrier and / or excipients to obtain a traditional Chinese medicine preparation.

[0012] Preferably, the preparation method includes: mixing peony bark and Cynanchum paniculatum with water, reflux extraction, and collecting the distillate; adding sodium chloride to the distillate, refrigerating, filtering, and drying to obtain paeonol; encapsulating paeonol with beta-cyclodextrin to obtain a paeonol inclusion complex; combining the residue after paeonol extraction with Rehmannia glutinosa, Forsythia suspensa, Paeonia lactiflora, Polygonum cuspidatum, Scutellaria baicalensis, Cimicifuga foetida, and Pueraria lobata, adding water and decocting, filtering, collecting the filtrate and concentrating under reduced pressure to a clear extract with a density of 1.10~1.20; and combining the paeonol inclusion complex and the concentrated extract with a carrier and / or excipients to obtain a traditional Chinese medicine preparation.

[0013] Fourthly, the present invention provides the use of the aforementioned traditional Chinese medicine composition, or the aforementioned traditional Chinese medicine preparation, or the traditional Chinese medicine preparation obtained by the aforementioned preparation method in the preparation of a medicament for the prevention or treatment of Kawasaki disease.

[0014] Preferably, the Kawasaki disease is of the heat-toxin and blood stasis type.

[0015] Beneficial effects:

[0016] This invention provides a traditional Chinese medicine composition, its preparation method, and its application. This composition is a pure traditional Chinese medicine preparation with the effects of clearing heat and cooling blood, promoting blood circulation, and detoxifying. It is suitable for Kawasaki disease (especially the heat-toxin and blood stasis type). The traditional Chinese medicine composition of this invention is prepared from the following raw materials in parts by weight: 8-30 parts Rehmannia glutinosa, 2-10 parts Forsythia suspensa, 2-10 parts Paeonia suffruticosa, 2-10 parts Paeonia lactiflora, 2-10 parts Polygonum cuspidatum, 2-10 parts Scutellaria baicalensis, 2-10 parts Cynanchum paniculatum, 1-5 parts Cimicifuga foetida, and 3-15 parts Pueraria lobata. Experimental results show that the traditional Chinese medicine composition provided by this invention has a good therapeutic effect on Kawasaki disease. It can significantly improve cardiac contractile function, increase left ventricular ejection fraction and fractional shortening rate; reduce coronary artery inflammation infiltration, intimal hyperplasia and myocardial damage, and reduce serum myocardial injury markers FABP and BNP levels; regulate systemic inflammatory response, reduce PCT, VEGF and CRP levels, promote platelet system recovery, and improve erythrocyte distribution width; effectively correct the abnormally high proportion of Th1 cells, showing multi-target and overall regulatory therapeutic characteristics. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in this invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be described below.

[0018] Figure 1 This is a schematic diagram of the process control flow for the traditional Chinese medicine composition (extracted preparation) of the present invention.

[0019] Figure 2 The pathological images of the heart and coronary artery tissues of Kawasaki disease mice in Example 4 of this invention (scale bar: CardiacMuscle 0.100mm; Coronary Artery 0.050mm; Aortic Root 0.200mm). Detailed Implementation

[0020] While ancient Chinese medical texts do not use the name Kawasaki disease, it is similar to "spotted rash" described in the *Treatise on the Causes and Symptoms of Various Diseases*: "Heat enters the stomach, and the heat, carrying toxins, accumulates in the stomach. The toxins then manifest in the muscles, causing red spots to appear and spread throughout the body." Some scholars believe this disease falls under the category of "febrile diseases," while others believe it should be classified as "epidemic rash" or "spotted rash." The herbal formula provided in this invention is derived from the clinical experience of the renowned traditional Chinese medicine master Lu Fang. It consists of Rehmannia glutinosa, Forsythia suspensa, Paeonia suffruticosa, Paeonia lactiflora, Polygonum cuspidatum, Scutellaria baicalensis, Cynanchum paniculatum, Cimicifuga foetida, and Pueraria lobata, with the effects of clearing heat and cooling blood, promoting blood circulation, and detoxifying. This invention uses this herbal formula to treat Kawasaki disease of the heat-toxin and blood stasis type, which aligns with the traditional Chinese medicine principle that the pathogenic heat toxins accumulate in the lungs and stomach.

[0021] Specifically, Rehmannia glutinosa has the effects of cooling the blood and eliminating spots, nourishing yin and generating fluids. Its taste is sweet and cold, specifically entering the blood level, clearing the heart and cooling the blood, and nourishing the blood vessels. This perfectly corresponds to the core pathogenesis of Kawasaki disease, which involves intense heat and toxicity, heat entering the blood level, depleting yin fluids, and damaging the blood vessels. In this invention, Rehmannia glutinosa can clear heat and toxicity from the qi, blood, and blood levels, improving acute symptoms of Kawasaki disease such as high fever, generalized rashes, chapped lips, and swollen lymph nodes. It can also cool the blood and disperse blood stasis, clearing heat stagnation in the blood and relieving redness, swelling, and peeling of the extremities. Simultaneously, it nourishes yin and protects fluids, nourishing the heart vessels. Targeting the pathogenesis of this disease—heat and toxicity damaging yin, impairing blood vessels, and easily leading to coronary artery stasis—it plays a key role in clearing heat, cooling the blood, nourishing yin, and protecting the blood vessels. It is the core medicine for Kawasaki disease characterized by simultaneous burning of qi and blood, and damage to yin and blood vessels. Furthermore, this invention uses Rehmannia glutinosa as the key herb, which aligns with the pathogenesis of Kawasaki disease in its acute phase. Rehmannia glutinosa is combined with Paeonia suffruticosa and Paeonia lactiflora, focusing on the four key aspects of heat toxicity, blood stasis, yin deficiency, and collateral damage. This approach cools the blood, disperses blood stasis, nourishes yin, and protects the collaterals, consistent with the comprehensive diagnosis and treatment of Kawasaki disease. Therefore, this invention heavily utilizes Rehmannia glutinosa and Forsythia suspensa as the principal herbs, Paeonia suffruticosa, Paeonia lactiflora, Pueraria lobata, and Polygonum cuspidatum as the assistant herbs, and Scutellaria baicalensis, Cynanchum paniculatum, and Cimicifuga foetida as adjuvant herbs. Together, these herbs achieve the effects of clearing heat, cooling the blood, promoting blood circulation, and detoxifying.

[0022] In one of the more specific embodiments provided by the present invention, the raw material ratio of the traditional Chinese medicine composition is as follows: 15 parts of Rehmannia glutinosa, 5 parts of Forsythia suspensa, 5 parts of Paeonia suffruticosa, 5 parts of Paeonia lactiflora, 5 parts of Polygonum cuspidatum, 5 parts of Scutellaria baicalensis, 5 parts of Cynanchum paniculatum, 2.5 parts of Cimicifuga foetida, and 7.5 parts of Pueraria lobata.

[0023] This invention does not specifically limit the source of the above-mentioned raw materials; any commercially available source in the art is acceptable. Preferably, the above-mentioned raw materials are processed using the following methods: Rehmannia glutinosa: remove impurities, wash, moisten, cut into thick slices, and dry; Forsythia suspensa: remove impurities; Paeonia suffruticosa: quickly wash, moisten, cut into thin slices, and sun-dry; Paeonia lactiflora: remove impurities, separate by size, wash, moisten thoroughly, cut into thick slices, and dry; Polygonum cuspidatum: remove impurities, wash, moisten thoroughly, cut into thick slices, and dry; Scutellaria baicalensis: remove impurities, boil in boiling water for 10 minutes, remove, moisten thoroughly, cut into thin slices, and dry; or steam for half an hour, remove, cut into thin slices, and dry (avoid direct sunlight); Cynanchum paniculatum: remove impurities, quickly wash, cut into sections, and air-dry; Cimicifuga foetida: remove impurities, soak briefly, wash, moisten thoroughly, cut into thick slices, and dry; Pueraria lobata: remove impurities, wash, moisten thoroughly, cut into thick slices, coarse shreds, or cubes, and sun-dry.

[0024] The present invention weighs the above-mentioned active pharmaceutical ingredients according to their weight parts; extracts them in an extraction solvent (water), then concentrates and dries them; and finally combines them with a carrier and / or excipients to obtain preparations such as granules, tablets, and capsules.

[0025] In one specific embodiment of the present invention, the preparation method is as follows: ① Weigh each raw medicinal material according to the weight proportions.

[0026] ② Take peony bark and cynanchum paniculatum, add 18-24 times the amount of drinking water and reflux for 2-5 hours to extract, and collect the distillate. Add 1-2% sodium chloride (by volume) to the distillate, refrigerate at 2-10℃, filter, and dry to obtain paeonol for later use.

[0027] ③ Take paeonol, encapsulate it with beta-cyclodextrin, and set aside.

[0028] ④ Take seven herbs including Rehmannia glutinosa, combine them with the above-mentioned dregs, add 10-14 times the amount of drinking water for these seven herbs and decoct them, then add 8-12 times the amount of drinking water for decoction, filter, combine the filtrates, concentrate under reduced pressure to a clear extract with a relative density of 1.10~1.20 (measured at 70-80℃), filter, and set aside.

[0029] ⑤ Take an appropriate amount of paeonol inclusion complex, dextrin, and sucralose, mix them with the above concentrated extract, and granulate to obtain the final product.

[0030] The composition provided by this invention is a pure traditional Chinese medicine preparation with a novel and original formula and a clear clinical positioning. It is suitable for Kawasaki disease (especially the heat-toxin and blood stasis type), exerts the effects of clearing heat and cooling blood, promoting blood circulation and detoxifying, and can directly target the core pathogenesis, fully exert the efficacy of the prescription, and has a definite curative effect.

[0031] The preparation method provided by this invention is simple, has few medicinal ingredients, and can be formulated into pharmaceutically acceptable dosage forms (such as tablets, capsules, granules, etc.). This invention preferably prepares it as granules, which are more convenient for clinical use, have a larger drug loading capacity, and are easy to store and carry.

[0032] The invention has low overall cost and is easily accepted by patients.

[0033] To make the above-mentioned objects, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to the examples in the specification.

[0034] Many specific details are set forth in the following description in order to provide a full understanding of the invention. However, the invention may also be practiced in other ways different from those described herein, and those skilled in the art can make similar extensions without departing from the spirit of the invention. Therefore, the invention is not limited to the specific embodiments disclosed below.

[0035] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "specific implementation," or "some specific implementations," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0036] The endpoints and any values ​​of the ranges disclosed in this specification are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of the various ranges, the endpoint values ​​of the various ranges and individual point values, and individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed herein.

[0037] In the embodiments provided in this specification, unless specific techniques or conditions are specified, the techniques or conditions described in the literature in this field, or the product instructions, shall be followed. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased from legitimate channels.

[0038] Example 1

[0039] This embodiment provides a traditional Chinese medicine composition for treating Kawasaki disease. The daily prescription amount of its raw materials, by weight, is as follows: 15g Rehmannia glutinosa, 5g Forsythia suspensa, 5g Paeonia suffruticosa, 5g Paeonia lactiflora, 5g Polygonum cuspidatum, 5g Scutellaria baicalensis, 5g Cynanchum paniculatum, 2.5g Cimicifuga foetida, and 7.5g Pueraria lobata.

[0040] This embodiment also provides a method for preparing the above-mentioned traditional Chinese medicine composition, the steps of which are as follows: (1) Take the prescribed amount of peony bark and cynanchum paniculatum, add 24 times the amount of drinking water, reflux extract at room temperature for 3.5 hours, and collect the distillate; add 1.5% of the distillate volume of sodium chloride to the distillate, refrigerate at 6℃; filter to collect the solid part, dry, and obtain paeonol for later use.

[0041] (2) Take paeonol and encapsulate it with 7.5 to 10 times the weight of beta-cyclodextrin to obtain paeonol inclusion complex for later use.

[0042] (3) Take the prescription amount of Rehmannia glutinosa and other seven herbs, combine them with the dregs of the peony bark and Cynanchum paniculatum extracted by reflux in step (1), add 12 times the amount of drinking water for the seven herbs and decoct for 1 hour, filter through 300 mesh; add 10 times the amount of drinking water for the seven herbs and decoct for 40 minutes, filter through 300 mesh, combine the filtrates; concentrate under reduced pressure to a clear extract with a relative density of 1.10~1.20 (measured at 70-80℃), filter through 300 mesh, and set aside.

[0043] (4) Take an appropriate amount of paeonol inclusion complex, concentrated paste, dextrin, and sucralose, mix and granulate.

[0044] In this embodiment, the total amount of paeonol prepared in step (1) is 120 mg; the total amount of inclusion complex prepared in step (2) is 0.9 g; the total amount of extract prepared in step (3) is 18.15 g; and the amount of dextrin used in step (4) is 12.82 g and the amount of sucralose used is 0.13 g.

[0045] Example 2

[0046] The difference between this embodiment and Embodiment 1 lies in the prescription: 8g of Rehmannia glutinosa, 2g of Forsythia suspensa, 2g of Paeonia suffruticosa, 2g of Paeonia lactiflora, 2g of Polygonum cuspidatum, 2g of Scutellaria baicalensis, 2g of Cynanchum paniculatum, 1g of Cimicifuga foetida, and 3g of Pueraria lobata.

[0047] Example 3

[0048] The difference between this embodiment and Embodiment 1 lies in the prescription: 25g Rehmannia glutinosa, 10g Forsythia suspensa, 10g Paeonia suffruticosa, 10g Paeonia lactiflora, 10g Polygonum cuspidatum, 10g Scutellaria baicalensis, 10g Cynanchum paniculatum, 4g Cimicifuga foetida, and 14g Pueraria lobata.

[0049] Comparative Example 1 This comparative example provides a traditional Chinese medicine composition with the following prescription: 10g Polygonum cuspidatum, 12g Cynanchum paniculatum, 10g Dictamnus dasycarpus, 10g Smilax glabra, 10g Scutellaria baicalensis, 10g Forsythia suspensa, 10g Paeonia suffruticosa, and 12g Paeonia lactiflora.

[0050] The preparation method of this comparative example of traditional Chinese medicine composition is as follows: Weigh the raw materials according to the prescribed dosage, then add 8 times the amount of water to each raw material, decoct for 40 minutes, filter through a 300-mesh sieve, add 6 times the amount of water to the residue, decoct for 30 minutes, filter through a 300-mesh sieve again, combine the decoctions, and concentrate to a density of 1.2 ± 0.05 (60℃) to obtain a concentrated paste. Dry under reduced pressure at 70℃ to obtain a dry paste. Pulverize the dry paste through a 60-mesh sieve to obtain an extract powder, add 12g of dextrin, mix well, granulate with 70% ethanol, and dry.

[0051] Example 4

[0052] This embodiment experimentally verifies the efficacy of the traditional Chinese medicine composition provided by the present invention.

[0053] In this embodiment, the experimental drugs are the traditional Chinese medicine composition of Example 1, the traditional Chinese medicine composition of Comparative Example 1, aspirin, and Lactobacillus casei cell wall extract (LCWE) (self-made).

[0054] The preparation method of Lactobacillus casei cell wall extract is as follows: Lactobacillus casei culture: Weigh 52.4g of MRS broth medium, heat and dissolve in 1000ml of distilled water, autoclave at 118℃ for 15min, cool and store at 4℃ for later use; Thaw Lactobacillus casei on crushed ice, and inoculate the thawed Lactobacillus casei into MRS broth medium in a clean bench, and culture anaerobically for 48h; Inoculate the Lactobacillus casei cultured for 48h into a total of 2L of new MRS broth medium for expansion culture, and culture anaerobically for 48h.

[0055] LCWE extraction: After anaerobic culture of *Lactobacillus casei* for 48 h, the supernatant was discarded by centrifugation, and the precipitate was retained. The precipitate was washed 6 times with PBS, and 10 volumes of 4% SDS were added to lyse the bacteria. The bacterial suspension containing 4% SDS was thoroughly resuspended by refluxing, and then placed on a shaker and incubated overnight at 250 rpm / min at room temperature. After overnight incubation, the precipitate was centrifuged at 10,000 rpm and 4°C for 40 min, and the supernatant was discarded. The bacterial precipitate was washed 8 times with PBS. Then, the precipitate was sequentially treated with 10 volumes of 250 μg / mL DNase I, RNase A, and trypsin for 4 h under the following conditions: shaker 250 rpm / min and 37°C. After each enzyme treatment, the precipitate was thoroughly washed twice with PBS buffer. The final digestion with trypsin was followed by 4 washes with PBS buffer. After washing, the precipitate was centrifuged at 10,000 rpm and 4°C for 40 min, and the supernatant was discarded. The bacterial pellet was resuspended in 4 times its volume of PBS buffer. The bacterial suspension was then subjected to ice-bath sonication to disrupt the cell walls at room temperature under the following conditions: amplitude 300W, sonication for 10 seconds, pause for 5 seconds, and sonication for 2.5 hours. The resulting suspension was observed to be a thin, milky white color and capable of dripping spontaneously. The suspension was then centrifuged at 1000 rpm at 4°C for 1 hour. The supernatant was collected and filtered through a 0.22 μm filter to obtain LCWE, which was aliquoted into 1.5 mL EP tubes and stored at -80°C for later use.

[0056] In this embodiment, the experimental animals were healthy male C57 mice aged 4-5 weeks. Housing conditions: room temperature 20-26℃, humidity 40-70%, and a 12h / 12h light / dark cycle.

[0057] In this embodiment, the experimental groups were: blank control group, model group, positive control group (aspirin, 15 mg / kg, prepared at 1.5 mg / mL), comparative example 1 treatment group (12.74 g crude drug / kg), example 1 low-dose treatment group (4.17 g crude drug / kg), example 1 medium-dose treatment group (8.34 g crude drug / kg), and example 1 high-dose treatment group (16.68 g crude drug / kg). The groups were randomly assigned according to the required sample size and the dosage was calculated based on mouse body weight. Each treatment group received the medication once daily via gavage at a fixed time for 28 consecutive days.

[0058] The experimental steps in this embodiment include: 1. Model Establishment: C57 mice were acclimatized for one week and then randomly divided into a normal control group, a model control group, a positive control group (aspirin 15 mg / kg), a comparative example 1 treatment group (12.74 g crude drug / kg), and Example 1 (test drug SLCQ) low, medium, and high dose groups (4.17 g crude drug / kg, 8.34 g crude drug / kg, and 16.68 g crude drug / kg). Except for the normal control group, all other groups received intraperitoneal injections of LCWE extract (0.5 mg / mouse) once daily for 28 consecutive days. The drug was administered simultaneously with model establishment, and mice were randomly assigned to groups based on the required sample size, with dosages calculated according to mouse body weight. Starting the day after LCWE injection, each treatment group received the drug via gavage once daily for 28 consecutive days. Three days after injection, mice were observed to be irritable, huddled together, had dull fur, and exhibited slight redness and swelling in the mouth and nose areas.

[0059] 2. Observation indicators: Cardiac function assessment, HE staining of the heart and coronary arteries, measurement of serum myocardial injury markers FABP and BNP, measurement of serum biochemical indicators VEGF, PCT, and CRP, and flow cytometry analysis of Th1 cell proportion.

[0060] 3. Experimental Results: (1) Effects of traditional Chinese medicine on cardiac function in Kawasaki disease mice.

[0061] Compared with the normal control group, the left ventricular ejection fraction (EF) and fractional shortening (FS) of the model group mice were significantly reduced (p<0.01), indicating that the Kawasaki disease model successfully induced severe damage to cardiac contractile function in mice. Compared with the model group, both the medium and high dose groups effectively reversed the deterioration of cardiac function indicators, with EF and FS values ​​significantly improved compared with the model group (p<0.01), and the improvement increased with increasing drug concentration. The high-dose group showed a greater degree of recovery in EF and FS than the traditional positive control drug aspirin. The results are shown in Table 1.

[0062] Table 1. Cardiac systolic function in Kawasaki disease mice (mean ± SD)

[0063] Note: ***P < 0.001, compared with the Control group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the Model group.

[0064] (2) Effects of traditional Chinese medicine on the histopathology of the heart and coronary arteries in Kawasaki disease mice.

[0065] Compared with the normal control group, the model group mice showed typical Kawasaki disease pathological changes, with extensive inflammatory cell infiltration in the myocardial tissue, myocardial cell swelling, disordered arrangement, and local necrosis; the coronary arteries were particularly significantly affected, showing inflammation of the entire vessel wall, degeneration and necrosis of the smooth muscle cells in the media, significant intimal hyperplasia, and luminal stenosis; inflammatory cell infiltration and vessel wall edema were also observed at the aortic root, accompanied by an early dilation trend.

[0066] Compared with the model group, the pathological changes in the tested traditional Chinese medicine group were significantly reduced. In the myocardial tissue, the extent of inflammatory cell infiltration was significantly reduced, and myocardial cell edema and necrosis were effectively curbed. In particular, the high-dose group significantly inhibited inflammatory infiltration of the vascular wall, reduced damage to the tunica media smooth muscle, and effectively alleviated intimal hyperplasia and coronary artery wall edema, thereby maintaining better vascular patency. Inflammatory infiltration and structural damage in the aorta were also significantly improved. The aspirin treatment group also showed clear effects in inhibiting vascular and myocardial inflammation, reducing intimal hyperplasia, and improving the structural integrity of the aorta. Results Figure 1 .

[0067] (3) Effects of traditional Chinese medicine on serum myocardial injury markers in Kawasaki disease mice.

[0068] Compared with the normal group, the serum FABP and BNP levels in the model group mice were significantly higher than those in the normal control group (p<0.01), indicating that the model mice had clear myocardial injury and cardiac stress. Compared with the model group rats, the low, medium and high doses of the tested traditional Chinese medicine group could effectively reduce the abnormal increase of serum myocardial injury markers. Among them, the medium and high doses of FABP and BNP levels were significantly lower than those in the model group (p<0.01), and showed a certain dose-dependent effect, with similar efficacy to the positive control group. The results are shown in Table 2.

[0069] Table 2. Serum myocardial injury marker levels in Kawasaki disease mice (mean ± SD)

[0070] Note: **P < 0.01, ***P < 0.001, compared with the Control group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the Model group.

[0071] (4) Effects of traditional Chinese medicine on hematological and inflammatory and endothelial function-related serum biochemical indicators in Kawasaki disease mice.

[0072] The erythrocyte sedimentation rate (ESR) in the model group did not increase significantly, and no significant change in ESR was observed after intervention with the tested traditional Chinese medicine. Platelet count (PLT) was significantly decreased in the model group (p < 0.01), while the medium- and high-dose groups of the tested traditional Chinese medicine significantly increased platelet count compared to the model group (p < 0.05). Aspirin, as an antiplatelet drug, does not decrease platelet count but rather reduces the probability of platelet aggregation, preventing platelet aggregation and thrombus formation, thereby reducing the occurrence of thrombosis.

[0073] The white blood cell count (WBC) in the model group showed a slight decreasing trend, but there were no significant differences between the groups. Among the red blood cell parameters, the red blood cell distribution width standard deviation (RDW-SD) in the model group was significantly increased (p < 0.01). Each dose group of the tested traditional Chinese medicine had the potential to reduce RDW-SD.

[0074] Procalcitonin (PCT) and C-reactive protein (CRP), as markers of severe systemic inflammation, were significantly elevated in the model group compared to the normal control group (p < 0.05). All doses of the tested traditional Chinese medicine effectively reduced PCT, while medium and high doses effectively reduced CRP, with no statistically significant difference compared to aspirin. Simultaneously, vascular endothelial growth factor (VEGF) levels were significantly elevated in the model group (p < 0.01). The tested traditional Chinese medicine effectively reversed the abnormal increase in VEGF, and the efficacy showed a dose-dependent effect. The improvement effect of the high-dose group was comparable to that of the aspirin group. Results are shown in Tables 3-6.

[0075] Table 3 Erythrocyte sedimentation rate and platelet count levels (mean ± SD)

[0076] Note: **P < 0.01, compared with the Control group; ##P < 0.01, compared with the Model group.

[0077] Table 4 Standard deviation of white blood cell count and red blood cell distribution width (mean ± SD)

[0078] Note: **P < 0.01, compared to the Control group.

[0079] Table 5. Levels of procalcitonin and vascular endothelial growth factor (mean ± SD)

[0080] Note: **P < 0.01, ***P < 0.001, compared with the Control group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the Model group.

[0081] Table 6. C-Reactive Protein Levels (mean ± SD)

[0082] Note: *P < 0.05, compared with the Control group; ##P < 0.01, ###P < 0.001, compared with the Model group.

[0083] (5) Effects of traditional Chinese medicine on the proportion of Th1 cells in Kawasaki disease mice Compared with the normal control group, the proportion of Th1 cells in the peripheral blood of mice in the model group was significantly increased (p<0.01). After drug treatment intervention, the tested traditional Chinese medicine group showed significant immunomodulatory effects. Specifically, the low, medium, and high dose groups could effectively reverse the abnormal increase of Th1 cells (p<0.001). Moreover, the regulatory effect of each dose group of the tested traditional Chinese medicine on the proportion of Th1 cells was comparable to that of the aspirin positive drug group (p<0.001). The results are shown in Table 7.

[0084] Table 7. Proportion of Th1 cells (mean ± SD)

[0085] Note: ***P < 0.001 compared to the Control group; ###P < 0.001 compared to the Model group.

[0086] 4. Conclusion: The tested traditional Chinese medicine (TCM) significantly improved cardiac contractile function, increased left ventricular ejection fraction and fractional shortening; reduced coronary artery inflammation, intimal hyperplasia, and myocardial damage; and decreased serum levels of myocardial injury markers FABP and BNP. Simultaneously, the TCM regulated systemic inflammatory responses, reduced PCT, VEGF, and CRP levels, promoted platelet system recovery, and improved erythrocyte distribution width. In terms of immune regulation, the TCM effectively corrected the abnormally high proportion of Th1 cells, demonstrating multi-target and holistic regulatory therapeutic characteristics. In conclusion, the tested TCM demonstrated clear efficacy in improving cardiac function, inhibiting vascular inflammation, regulating immune imbalance, and restoring multi-system function, providing solid preclinical pharmacodynamic evidence for its clinical application.

[0087] The embodiments described above are merely illustrative of several implementations of the present invention, designed to facilitate a detailed understanding of the technical solutions of the present invention. However, they should not be construed as limiting the scope of patent protection. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition, characterized in that, The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 8-30 parts of Rehmannia glutinosa, 2-10 parts of Forsythia suspensa, 2-10 parts of Paeonia suffruticosa, 2-10 parts of Paeonia lactiflora, 2-10 parts of Polygonum cuspidatum, 2-10 parts of Scutellaria baicalensis, 2-10 parts of Cynanchum paniculatum, 1-5 parts of Cimicifuga foetida, and 3-15 parts of Pueraria lobata.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 8-15 parts Rehmannia glutinosa, 2-5 parts Forsythia suspensa, 2-5 parts Paeonia suffruticosa, 2-5 parts Paeonia lactiflora, 2-5 parts Polygonum cuspidatum, 2-5 parts Scutellaria baicalensis, 2-5 parts Cynanchum paniculatum, 1-2.5 parts Cimicifuga foetida, and 3-7.5 parts Pueraria lobata.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 15 parts Rehmannia glutinosa, 5 parts Forsythia suspensa, 5 parts Paeonia suffruticosa, 5 parts Paeonia lactiflora, 5 parts Polygonum cuspidatum, 5 parts Scutellaria baicalensis, 5 parts Cynanchum paniculatum, 2.5 parts Cimicifuga foetida, and 7.5 parts Pueraria lobata.

4. A traditional Chinese medicine preparation, characterized in that, It is made alone from the traditional Chinese medicine composition according to any one of claims 1-3, or from the traditional Chinese medicine composition according to any one of claims 1-3 with a pharmaceutically acceptable carrier and / or excipient.

5. The traditional Chinese medicine preparation according to claim 4, characterized in that, The dosage forms of the traditional Chinese medicine preparations are tablets, pills, capsules, granules, powders, ointments, oral liquids, or mixtures.

6. The method for preparing the traditional Chinese medicine preparation according to claim 4 or 5, characterized in that, Includes the following steps: S1. Weigh each raw material according to the weight parts; S2. Place the raw material in the extraction solvent, separate, and obtain the extract; S3. Combine the extract with a carrier and / or excipients to obtain a traditional Chinese medicine preparation.

7. The preparation method according to claim 6, characterized in that, include: Mix peony bark and cynanchum paniculatum with water, reflux to extract, and collect the distillate; Sodium chloride was added to the distillate, and the mixture was refrigerated, filtered, and dried to obtain paeonol. Paeonol was then encapsulated with beta-cyclodextrin to obtain a paeonol inclusion complex. The residue after paeonol extraction was combined with Rehmannia glutinosa, Forsythia suspensa, Paeonia lactiflora, Polygonum cuspidatum, Scutellaria baicalensis, Cimicifuga foetida, and Pueraria lobata, decocted with water, filtered, and the filtrate was collected and concentrated under reduced pressure to a density of 1.10-1.20 to obtain a clear extract. The paeonol inclusion complex and the concentrated extract were combined with a carrier and / or excipients to obtain a traditional Chinese medicine preparation.

8. The use of the traditional Chinese medicine composition according to any one of claims 1-3, or the traditional Chinese medicine preparation according to claim 4 or 5, or the traditional Chinese medicine preparation prepared by the preparation method according to any one of claims 6-7 in the preparation of a drug for the prevention or treatment of Kawasaki disease.

9. The application according to claim 8, characterized in that, The Kawasaki disease described is of the heat-toxin and blood stasis type.

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