Preparation method and application of sedum aizoon and gynura procumbens plant composite powder

Abstract

The invention provides a preparation method and application of sedum aizoon and gynura procumbens plant composite powder, and the sedum aizoon and gynura procumbens plant composite powder comprises the following raw materials in parts by weight: 320-370 parts of sedum aizoon powder, 130-160 parts of gynura procumbens powder, 70-80 parts of oat powder, 45-58 parts of soybean milk powder, 25-32 parts of coconut milk powder, 10-19 parts of tartary buckwheat powder, 10-16 parts of barley seedling powder and 15-19 parts of crystalline fructose. And 9 to 12 parts of beta-cyclodextrine. Through synergistic fermentation of eurotium cristatum, lactobacillus plantarum and nano sparkling water and in combination with a nano gas-assisted drying technology, the retention efficiency of active ingredients and basic nutritional ingredients in the sedum aizoon and gynura procumbens plant composite powder is remarkably improved, and the powder caking rate is reduced. The prepared sedum aizoon and gynura procumbens plant composite powder has a remarkable improvement or treatment effect on atherosclerosis, and an innovative solution is provided for development of products for preventing and treating atherosclerosis through the innovative formula and process of the sedum aizoon and gynura procumbens plant composite powder.

Description

Technical Field

[0001] This invention belongs to the field of fermentation technology, specifically relating to a method for preparing a compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* and its application. Background Technology

[0002] Sedum aizoon is a perennial herb belonging to the genus Sedum in the family Crassulaceae. Also known as Sedum sarmentosum or Sedum dasycarpus, it is distributed in Northeast, North, and East China. It is a plant used for both medicinal and edible purposes, being easy to cultivate with a high survival rate. It is also rich in various active ingredients, including flavonoids, alkaloids, and phenolic substances, possessing multiple physiological functions such as lowering blood pressure and lipids, detoxification, and hemostasis. It is mainly used to treat hematochezia, purpura, palpitations, and insomnia. Gynostemma pentaphyllum is a perennial herb belonging to the genus Gynostemma in the family Asteraceae. Also known as Sedum sarmentosum or Gynostemma pentaphyllum, it is distributed in Southeast Asian countries such as Thailand, Indonesia, and Vietnam. In my country, it is mainly distributed in Guangdong, Jiangxi, Hainan, and Guizhou. Gynostemma pentaphyllum is also a plant used for both medicinal and edible purposes, rich in polysaccharides, flavonoids, phenolic acids, terpenes, and fatty acids, possessing pharmacological effects such as anti-inflammatory and antipyretic properties, promoting blood circulation and removing blood stasis, reducing swelling and relieving pain, and protecting the liver and detoxifying. Studies have found that *Gynura divaricata* has potential therapeutic effects on various diseases, including cancer, hypertension, and diabetes. Plants that are both food and medicine are considered a low-cost, safe, and effective treatment method. *Gynura divaricata* and *Gynura divaricata* possess both medicinal and edible value, are rich in various active ingredients, and have significant development potential in the functional food and pharmaceutical fields.

[0003] Traditional processing methods suffer from low extraction efficiency and are prone to degradation of heat-sensitive components, limiting their bioavailability and application effects. In existing technologies, microbial fermentation, through the synergistic fermentation of specific strains, can decompose plant cell wall structures, promoting the release and transformation of active ingredients (such as flavonoids, polysaccharides, and saponins) and generating new metabolites, thereby enhancing their physiological activity. However, simple fermentation suffers from long cycles, low efficiency, and easy clumping during drying, affecting the stability and efficacy of the final product. Therefore, developing a new processing technology that balances extraction efficiency and component activity is of significant research value. Summary of the Invention

[0004] Technical problem to be solved: Based on the above technical problems, the first objective of this invention is to provide a compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng*.

[0005] The second objective of this invention is to provide a method for preparing a compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng*.

[0006] The third objective of this invention is to provide the application of a compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* in the preparation of pharmaceutical formulations for improving or treating atherosclerosis.

[0007] Technical solution: A method for preparing a compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng*, comprising the following steps:

[0008] S1. Activation of bacterial strains: The *Aurotriarcha* strain preserved on slant culture was inoculated into PDA solid medium and cultured. The mycelia were scraped and transferred to sterile water and shaken to obtain a suspension of *Aurotriarcha*. The *Lactobacillus plantarum* glycerol cryopreservation solution was inoculated into MRS liquid medium and cultured with shaking to obtain a suspension of *Lactobacillus plantarum*.

[0009] S2. Preparation of compound microbial agent: Mix the bacterial suspensions of *Aspergillus cristatus* and *Lactobacillus plantarum* evenly to prepare a compound microbial agent;

[0010] S3. Preparation of nanobubble water: Sterile water is injected into a nanobubble generator and treated with O2 to obtain nanobubble water;

[0011] S4. Raw material pretreatment: The powder of *Heliotropium indicum*, the powder of *Chrysanthemum indicum*, the crystalline fructose and nano-bubble water are sheared in a high-speed mixing tank, transferred to a mixing tank, and then oat flour, soy milk powder, coconut milk powder, buckwheat flour, barley grass powder and cyclodextrin are added and stirred evenly. After shearing and homogenization, the fermentation substrate is obtained.

[0012] S5. Fermentation: The fermentation substrate is transferred to a fermentation tank for sterilization. After cooling, a compound bacterial agent is added and stirred for fermentation. O2 nano gas is introduced in the first stage of fermentation using nano bubble technology, and N2 nano gas is introduced in the second stage of fermentation. After stirring and fermentation is completed, the fermentation is allowed to stand. The fermentation liquid is homogenized and filtered to obtain the filtrate.

[0013] S6. Drying: Nano gas is continuously introduced into the filtrate, food-grade silica is added and stirred evenly, and then spray-dried to obtain the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum* and *Panax notoginseng*.

[0014] As preferred strains, *Aspergillus cristatus* strain number CICC 2099 and *Lactobacillus plantarum* strain WCSF1 are used.

[0015] Preferably, the temperature for culturing *Eurotium cristatum* in step S1 is 25-28°C, and the culturing time is 3-5 days.

[0016] Preferably, the temperature for culturing *Lactobacillus plantarum* in step S1 is 35-37°C, and the culturing time is 12-24 h.

[0017] Preferably, the volume ratio of the *Aspergillus cristatus* and *Lactobacillus plantarum* suspension in the compound bacterial agent in step S1 is 2-5:1-3.

[0018] Preferably, the viable count of *Aspergillus cristatus* in the compound bacterial agent in step S1 is (3.5~4.5) × 10⁻⁶. 6CFU / mL, viable count of Lactobacillus plantarum was (2.8~3.6)×10⁻¹⁰. 6 CFU / mL.

[0019] Preferably, the diameter of the nanobubbles in the nanobubble water in step S3 is 50~200 nm.

[0020] Preferably, the O2 treatment time in step S3 is 30-35 minutes.

[0021] Preferably, the dissolved oxygen content of the nanobubble water in step S3 is 25~35 mg / L.

[0022] Preferably, the raw materials in step S4 include 320-370 parts of *Hedyotis diffusa* powder, 130-160 parts of *Chrysanthemum indicum* and *Panax notoginseng* powder, 70-80 parts of oat flour, 45-58 parts of soy milk powder, 25-32 parts of coconut milk powder, 10-19 parts of buckwheat flour, 10-16 parts of barley grass powder, 15-19 parts of crystalline fructose, and 9-12 parts of β-cyclodextrin.

[0023] Preferably, the shearing time of the high-speed mixing tank in step S4 is 5-9 min.

[0024] Preferably, the shearing time in the mixing tank in step S4 is 12-18 min.

[0025] Preferably, the homogenization pressure in step S4 is 35~45 MPa.

[0026] Preferably, the sterilization temperature in step S5 is 120~125℃, and the sterilization time is 5~15 s.

[0027] Preferably, in step S5, the inoculation amount of the compound microbial agent is 1-3%, and the stirring speed is 100-200 rpm.

[0028] Preferably, the temperature of the first stage of fermentation in step S5 is 25~28℃, and the fermentation time is 4~6 days.

[0029] Preferably, the fermentation temperature in the second stage of step S5 is 35~37℃, and the fermentation time is 5~7 days.

[0030] Preferably, the nano gas in step S5 includes, but is not limited to, O2, H2 and N2.

[0031] Preferably, the amount of food-grade silica added in step S5 is 0.5~1 wt%.

[0032] The above-mentioned compound powder of *Hedyotis diffusa*, *Chrysanthemum indicum*, and *Panax notoginseng* is used in the preparation of drug formulations for improving or treating atherosclerosis.

[0033] The dosage forms of the aforementioned drug preparations that have the function of improving or treating atherosclerosis are tablets, powders, granules, liquids, and capsules.

[0034] Beneficial effects:

[0035] 1. This invention combines microbial fermentation with nanobubble technology. The cavitation effect and interfacial activity of nanobubble water can effectively disrupt the plant cell wall structure, promote the release of active ingredients such as flavonoids and polysaccharides, and increase oxygen mass transfer efficiency and promote the dissolution of hydrophobic components. At the same time, the O2 slowly released by the nanobubbles can optimize the aerobic fermentation stage of *Aspergillus cristatus* while avoiding the anaerobic inhibition of *Lactobacillus plantarum*.

[0036] 2. This invention utilizes nano-gas-assisted drying technology, fully leveraging the high specific surface area, strong permeability, and unique gas-liquid interface effect of nano-gases. The nano-gas generates micro-turbulence in the liquid, disrupting surface tension and enhancing the diffusion efficiency of moisture from the material's interior to the surface, thus lowering the drying temperature and increasing drying efficiency. Simultaneously, the energy released when the bubbles burst promotes localized moisture evaporation. The nano-gas adhering to the surface of solid particles forms a physical isolation layer, reducing direct contact between particles and lowering the probability of agglomeration. Furthermore, the electrostatic repulsion of the bubbles reduces van der Waals force adsorption between particles. During the drying stage, the micro-explosion effect of the nano-gas prevents particle aggregation, improves powder flowability, and the low-temperature drying conditions maximize the protection of heat-sensitive active ingredients.

[0037] 3. This invention establishes a novel integrated extraction-fermentation-drying process with synergistic effects of physical and biological processes. Through the synergistic effect of multiple processes, it not only significantly improves the retention rate of bioactive components in the whole powder of *Gynostemma pentaphyllum* and *Chrysanthemum indicum*, but also improves the flowability and stability of the powder. This achieves multiple optimizations, including improved fermentation efficiency, improved drying quality, and improved storage stability, resulting in a final product that exhibits superior bioactivity in improving or treating atherosclerosis. Attached Figure Description

[0038] Figure 1 The total polysaccharide content of the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* in Examples 1-7 and Comparative Examples 1-5;

[0039] Figure 2 The total flavonoid content of the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* in Examples 1-7 and Comparative Examples 1-5;

[0040] Figure 3 The total triterpenoid content of the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* in Examples 1-7 and Comparative Examples 1-5;

[0041] Figure 4The total protein content of the compound powder of *Hedyotis diffusa*, *Chrysanthemum indicum* and *Panax notoginseng* in Examples 1-7 and Comparative Examples 1-5;

[0042] Figure 5 The total fat content of the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum* and *Panax notoginseng* in Examples 1-7 and Comparative Examples 1-5;

[0043] Figure 6 The agglomeration rate of the compound powder of *Hedyotis diffusa*, *Chrysanthemum indicum* and *Panax notoginseng* in Examples 1-7 and Comparative Examples 6-7;

[0044] Figure 7 This is a schematic diagram of zebrafish heart blood cell staining in Example 8;

[0045] Figure 8 Example 8 illustrates the intervention effect of the compound powder of *Hedyotis diffusa*, *Chrysanthemum indicum*, and *Panax notoginseng* on atherosclerosis. Detailed Implementation

[0046] The present invention will be further described below with reference to embodiments. These embodiments are illustrative of the present invention, but the present invention is not limited to these embodiments:

[0047] The strain of *Eurotium cristatum* used in the following examples has the accession number CICC 2099 and was purchased from Ningbo Mingzhou Biotechnology Co., Ltd., catalog number B00016; *Lactobacillus plantarum* WCSF1 was purchased from Hangzhou Baosai Biotechnology Co., Ltd.

[0048] Example 1

[0049] This embodiment describes the preparation of a compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng*, including the following steps:

[0050] S1. Activation of bacterial strains: *Aspergillus cristatus* preserved on slant culture was inoculated into PDA solid medium and incubated upside down at 28°C for 5 days. Mycelia were scraped and transferred to 50 mL of sterile water and cultured with shaking at 200 rpm for 1 h. The mycelia were counted using a hemocytometer and then used for later use. *Lactobacillus plantarum* WCSF1 glycerol cryopreservation solution was inoculated into MRS liquid medium and cultured with shaking at 37°C and 280 rpm for 12 h. The absorbance at 600 nm was measured using a spectrophotometer.

[0051] S2. Preparation of compound microbial agent: Mix the bacterial suspensions of *Aspergillus cristatus* and *Lactobacillus plantarum* at a ratio of 2:1 (v:v) to obtain the compound microbial agent;

[0052] S3. Preparation of nanobubble water: Sterile water is injected into a nanobubble generator and treated with O2 (purity ≥99%) for 30 min. The diameter of the nanobubbles is 50 nm and the dissolved oxygen content is 25 mg / L, thus obtaining nanobubble water.

[0053] S4. Raw material pretreatment: Take 340 parts of Herba Sedum powder, 150 parts of Chrysanthemum indicum powder, and 17 parts of crystalline fructose and add them to 500 parts of nano-bubble water. Shear in a high-speed mixing tank for 5 min, transfer to a mixing tank, and then add 80 parts of oat flour, 58 parts of soy milk powder, 32 parts of coconut milk powder, 19 parts of buckwheat flour, 16 parts of barley grass powder and 10 parts of cyclodextrin. After uniform stirring, shear for 15 min and homogenize at 35 MPa to obtain the fermentation substrate.

[0054] S5. Fermentation: The fermentation substrate was transferred to a fermenter, sterilized at 120℃ for 15 s, cooled to 25℃, and then 3% of a compound bacterial agent was added. O2 nano gas with a flow rate of 0.5 L / min was introduced, and the mixture was stirred and fermented at 200 rpm and 28℃ for 5 days. After the first stage of fermentation, N2 nano gas with a flow rate of 0.2 L / min was introduced, and the mixture was stirred and fermented at 200 rpm and 37℃ for 6 days. After the second stage of fermentation, the mixture was allowed to stand for 2 days. After the fermentation was completed, the fermentation liquid was homogenized at 40 MPa and then filtered to obtain a filtrate with a solid content of 10%.

[0055] S6. Drying: N2 nano-gas was continuously introduced into the filtrate for 20 min, with a bubble diameter of 50 nm and a gas flow rate of 0.5 L / min. 0.5 wt% food-grade silica was added and stirred evenly before spray drying to obtain the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum* and *Panax notoginseng*.

[0056] Example 2

[0057] The difference between this embodiment and Embodiment 1 is that the raw materials in this embodiment are 330 parts of *Hedyotis diffusa* powder, 140 parts of *Chrysanthemum indicum* and *Panax notoginseng* powder, 78 parts of oat flour, 55 parts of soy milk powder, 28 parts of coconut milk powder, 15 parts of buckwheat flour, 18 parts of barley grass powder, 16 parts of crystalline fructose, 10 parts of β-cyclodextrin, and 540 parts of nano-bubble water.

[0058] Example 3

[0059] The difference between this embodiment and Embodiment 1 is that the raw materials in this embodiment are 370 parts of *Hedyotis diffusa* powder, 160 parts of *Chrysanthemum indicum* and *Panax notoginseng* powder, 88 parts of oat flour, 62 parts of soy milk powder, 36 parts of coconut milk powder, 24 parts of buckwheat flour, 21 parts of barley grass powder, 20 parts of crystalline fructose, 15 parts of β-cyclodextrin, and 600 parts of nano-bubble water.

[0060] Example 4

[0061] The difference between this embodiment and Embodiment 1 is that the inoculation amount of the compound microbial agent in this embodiment is 1%.

[0062] Example 5

[0063] The difference between this embodiment and Embodiment 1 is that the inoculation amount of the compound microbial agent in this embodiment is 2%.

[0064] Example 6

[0065] The difference between this embodiment and Embodiment 1 is that the gas flow rate of O2 nano gas in this embodiment is 0.2 L / min, and the gas flow rate of N2 nano gas is 0.1 L / min.

[0066] Example 7

[0067] The difference between this embodiment and Embodiment 1 is that in this embodiment, the flow rate of O2 nano-gas is 0.7 L / min, and the flow rate of N2 nano-gas is 0.5 L / min.

[0068] To further illustrate the technical effects of the present invention, a comparative example is also provided, as follows:

[0069] Comparative Example 1

[0070] The difference between this comparative example and Example 1 is that distilled water is used instead of nano-bubble water in this comparative example.

[0071] Comparative Example 2

[0072] The difference between this comparative example and Example 1 is that no bacterial agent is inoculated in this comparative example.

[0073] Comparative Example 3

[0074] The difference between this comparative example and Example 1 is that in this comparative example, *Eurotium cristatum* was inoculated alone.

[0075] Comparative Example 4

[0076] The difference between this comparative example and Example 1 is that Lactobacillus plantarum WCSF1 was inoculated alone in this comparative example.

[0077] Comparative Example 5

[0078] The difference between this comparative example and Example 1 is that nano-gas is not introduced during the fermentation process in this comparative example.

[0079] Comparative Example 6

[0080] The difference between this comparative example and Example 1 is that nanotechnology is not used during drying in this comparative example; the fermentation broth is directly spray-dried after filtration.

[0081] Comparative Example 7

[0082] The difference between this comparative example and Example 1 is that no silicon dioxide is added in this comparative example.

[0083] Table 1 Microbial test results

[0084]

[0085] As shown in Table 1, the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum* and *Panax notoginseng* prepared by this invention was free from other microbial contamination during the preparation process.

[0086] like Figures 1-5 As shown, compared with ordinary distilled water fermentation (Comparative Example 1), the total polysaccharide content, total flavonoid content, total triterpenoid content, protein content, and fat content of the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* fermented with nanobubble water (Examples 1-7) were significantly increased. The cavitation effect of nanobubbles promoted cell wall disruption, and the high specific surface area and surface charge could increase oxygen mass transfer efficiency, promoting the release and dissolution of active ingredients. Compared with the uninoculated microbial agent (Comparative Example 2), the total polysaccharide, total flavonoid, total triterpenoid, protein, and fat contents of the compound microbial agent inoculated with *Aspergillus cristatus* and *Lactobacillus plantarum* (Examples 1-7), the compound microbial agent inoculated with *Aspergillus cristatus* alone (Comparative Example 3), and the compound microbial agent inoculated with *Lactobacillus plantarum* alone (Comparative Example 4) were significantly increased. Although the contents of active ingredients and basic nutrients were also significantly increased by inoculating with *Aspergillus cristatus* alone (Comparative Example 3) or *Lactobacillus plantarum* alone (Comparative Example 4), their effects were significantly lower than those of the compound microbial agent treatment group (Examples 1-7). In addition, the inoculation amount of the compound microbial agent had a significant impact on the accumulation of active ingredients (Examples 1 and Examples 4-5), indicating that the fermentation of the compound microbial agent of *Aspergillus cristatus* and *Lactobacillus plantarum* can more effectively promote the dissolution and transformation of plant active ingredients and basic nutrients. Compared with the fermentation without the introduction of nano-gas (Comparative Example 5), the total polysaccharide, total flavonoid, total triterpenoid, protein and fat content of the compound powder of *Gynostemma pentaphyllum* and *Chrysanthemum indicum* with the introduction of nano-gas (Examples 1, 6-7) were significantly increased.

[0087] like Figure 6 As shown, compared with the absence of nano-gas-assisted drying technology (Comparative Example 6) or the absence of anti-caking agent silica (Comparative Example 7), Examples 1-7 showed a significant reduction in the agglomeration rate of the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum* and *Panax notoginseng* through the synergistic effect of nano-gas-assisted drying technology and silica.

[0088] Example 8

[0089] This embodiment describes the effect of the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* prepared in Example 1 on improving atherosclerosis in zebrafish, including the following steps:

[0090] S1. Wild-type AB strain zebrafish were divided into groups of 10 zebrafish embryos that were 3 days old after fertilization in each group. These groups were the normal control group, the model control group, the positive control group, and the sample treatment group. The positive control group was Shengxue Ning tablets, and the sample treatment group was the compound powder of Yangxincao Pingwoju Sanqi prepared in Example 1.

[0091] S2. Model control group: Zebrafish embryos were transferred to 3 cm culture dishes, and 4.999 mL of fish embryo culture medium and 1 μL of 10 mmol / L phenylhydrazine solution were added;

[0092] Positive control group: Zebrafish embryos were transferred to 3 cm culture dishes and 4.999 mL of fish embryo culture medium, 1 μL of 10 mmol / L phenylhydrazine solution and 500 μg / mL Shengxuening tablets were added;

[0093] Sample processing group: Zebrafish embryos were transferred to 3 cm culture dishes, and 4.999 mL of fish embryo culture medium, 1 μL of 10 mmol / L phenylhydrazine solution and 500 μg / mL of herbal compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum* and *Panax notoginseng* were added.

[0094] S3. Perform blood cell-specific staining on whole zebrafish and analyze the signal intensity of blood cells.

[0095] like Figures 7-8 As shown, the blank control group of zebrafish exhibited normal cardiac blood flow and no thrombus formation. Compared to the blank control group, the staining area in the model control group was significantly reduced, indicating the successful establishment of the atherosclerosis model. Compared to the model control group, the staining area in the sample test group was significantly increased, demonstrating that the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* prepared in this invention has the effect of improving atherosclerosis.

[0096] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A method for preparing a compound powder of *Hedyotis diffusa*, *Chrysanthemum indicum*, and *Panax notoginseng*, characterized in that... Includes the following steps: S1. Activation of bacterial strains: The *Aurotriarcha* strain preserved on slant culture was inoculated into PDA solid medium and cultured. The mycelia were scraped and transferred to sterile water and shaken to obtain a suspension of *Aurotriarcha*. The *Lactobacillus plantarum* glycerol cryopreservation solution was inoculated into MRS liquid medium and cultured with shaking to obtain a suspension of *Lactobacillus plantarum*. S2. Preparation of compound microbial agent: Mix the bacterial suspensions of *Aspergillus cristatus* and *Lactobacillus plantarum* evenly to prepare a compound microbial agent; S3. Preparation of nanobubble water: Sterile water is injected into a nanobubble generator and treated with O2 to obtain nanobubble water; S4. Raw material pretreatment: The powder of *Heliotropium indicum*, the powder of *Chrysanthemum indicum*, the crystalline fructose and nano-bubble water are sheared in a high-speed mixing tank, transferred to a mixing tank, and then oat flour, soy milk powder, coconut milk powder, buckwheat flour, barley grass powder and cyclodextrin are added and stirred evenly. After shearing and homogenization, the fermentation substrate is obtained. S5. Fermentation: The fermentation substrate is transferred to a fermentation tank for sterilization. After cooling, a compound bacterial agent is added and stirred for fermentation. O2 nano gas is introduced in the first stage of fermentation using nano bubble technology, and N2 nano gas is introduced in the second stage of fermentation. After stirring and fermentation is completed, the fermentation is allowed to stand. The fermentation liquid is homogenized and filtered to obtain the filtrate. S6. Drying: Nano gas is continuously introduced into the filtrate, food-grade silica is added and stirred evenly, and then spray-dried to obtain the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum* and *Panax notoginseng*.

2. The preparation method according to claim 1, characterized in that: In step S1, the temperature for culturing *Aspergillus cristatus* is 25-28℃, and the culturing time is 3-5 days; the temperature for culturing *Lactobacillus plantarum* is 35-37℃, and the culturing time is 12-24 hours.

3. The preparation method according to claim 1, characterized in that: In step S2, the volume ratio of *Aurogonium cristatum* and *Lactobacillus plantarum* suspensions in the compound microbial agent is 2-5:1-3; the viable count of *Aurogonium cristatum* in the compound microbial agent is (3.5-4.5) × 10⁻⁶. 6 CFU / mL, viable count of Lactobacillus plantarum was (2.8~3.6)×10⁻¹⁰. 6 CFU / mL.

4. The preparation method according to claim 1, characterized in that: In step S3, the diameter of the nanobubbles in the nanobubble water is 50~200 nm; the O2 treatment time is 30~35 min; and the dissolved oxygen content of the nanobubble water is 25~35 mg / L.

5. The preparation method according to claim 1, characterized in that: The raw materials in step S4 include 320-370 parts of *Hedyotis diffusa* powder, 130-160 parts of *Chrysanthemum indicum* and *Panax notoginseng* powder, 70-80 parts of oat flour, 45-58 parts of soy milk powder, 25-32 parts of coconut milk powder, 10-19 parts of buckwheat flour, 10-16 parts of barley grass powder, 15-19 parts of crystalline fructose, and 9-12 parts of β-cyclodextrin.

6. The preparation method according to claim 1, characterized in that: In step S4, the shearing time in the high-speed mixing tank is 5-9 min; the shearing time in the batching tank is 12-18 min; and the homogenization pressure is 35-45 MPa.

7. The preparation method according to claim 1, characterized in that: In step S5, the sterilization temperature is 120-125℃ and the sterilization time is 5-15 seconds; the inoculation amount of the compound microbial agent is 1-3%; the stirring speed is 100-200 rpm; the fermentation temperature in the first stage is 25-28℃ and the fermentation time is 4-6 days; the fermentation temperature in the second stage is 35-37℃ and the fermentation time is 5-7 days.

8. The preparation method according to claim 1, characterized in that: The nano-gas in step S6 includes, but is not limited to, O2, H2 and N2; the amount of food-grade silica added is 0.5~1 wt%.

9. The use of the compound powder of *Gynostemma pentaphyllum*, *Chrysanthemum indicum*, and *Panax notoginseng* prepared by any one of claims 1 to 8 in the preparation of pharmaceutical preparations for improving or treating atherosclerosis.

10. The application according to claim 9, characterized in that: The dosage form of the drug preparation having the function of improving or treating atherosclerosis is tablet, powder, granule, liquid or capsule.

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