Ginsenoside composition for accelerating acetaldehyde metabolism as well as preparation method and application of ginsenoside composition

Abstract

The invention provides a ginsenoside composition for accelerating acetaldehyde metabolism as well as a preparation method and application thereof, and particularly belongs to the technical field of medicine preparation. The invention provides a ginsenoside composition. The ginsenoside composition is prepared from the following components in parts by weight: 0.5 to 25 parts of Rg5, 0.5 to 20 parts of Rk1, 0.5 to 15 parts of Rh4, 0.5 to 10 parts of Rk3 and 0.5 to 10 parts of 20 (S)-Rg3. The ginsenoside composition can improve the enzymatic activity of acetaldehyde dehydrogenase, accelerate acetaldehyde metabolism and accelerate removal of acetaldehyde in a human body, and is beneficial to reduction of liver injury, protection of a cardiovascular system, protection of a nervous system, reduction of inflammatory response and improvement of the overall metabolic function.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparation technology, specifically relating to a ginsenoside composition that accelerates acetaldehyde metabolism, its preparation method, and its application. Background Technology

[0002] Acetaldehyde is a toxic compound that is produced in the human body through normal metabolic pathways such as alcohol, pyruvate, threonine metabolism, and glycolysis. In the liver, it is catalyzed by aldehyde dehydrogenase (ALDH) to form non-toxic acetic acid, which is then eliminated. Acetaldehyde poses various health risks, including damage to the liver, kidneys, cardiovascular system, and immune system; chronic inflammation; neurological disorders; and cancer risk. Long-term excessive alcohol consumption leads to a large amount of alcohol being catalyzed by alcohol dehydrogenase (ADH) in the liver to form acetaldehyde. Limited ALDH enzyme activity leads to acetaldehyde accumulation, subsequently causing alcoholic liver disease (ALD). Endogenous acetaldehyde is also produced during glycolysis, pyruvate metabolism, threonine metabolism, and intestinal microbial metabolism. Therefore, even in non-drinkers, slow acetaldehyde metabolism and delayed elimination can cause harm. Currently, there are no effective drugs for acetaldehyde elimination. Therefore, a drug that can effectively accelerate the metabolism of acetaldehyde in the body is needed to protect the body from its damage. Summary of the Invention

[0003] The purpose of this invention is to provide a ginsenoside composition for accelerating acetaldehyde metabolism, its preparation method, and its application. The ginsenoside composition of this invention can increase the enzyme activity of acetaldehyde dehydrogenase, accelerate acetaldehyde metabolism, and speed up the clearance of acetaldehyde from the human body.

[0004] The present invention provides a ginsenoside composition comprising the following components in parts by weight: 0.5-25 parts Rg5, 0.5-20 parts Rk1, 0.5-15 parts Rh4, 0.5-10 parts Rk3 and 0.5-10 parts 20(S)-Rg3.

[0005] The present invention also provides the application of the ginsenoside composition described above in the preparation of products that enhance the enzyme activity of acetaldehyde dehydrogenase or in the preparation of products that accelerate acetaldehyde metabolism.

[0006] Preferably, the product that accelerates acetaldehyde metabolism includes products that have one or more functions such as protecting the liver, protecting the cardiovascular system, protecting the nervous system, and anti-inflammation.

[0007] The present invention also provides a ginsenoside composition comprising the following components in parts by weight: 0.5-3 parts Rg6, 0.5-4 parts Rg4, 0.5-8 parts 20(R)-Rg3, 0.5-25 parts Rg5, 0.5-20 parts Rk1, 0.5-15 parts Rh4, 0.5-10 parts Rk3 and 0.5-10 parts 20(S)-Rg3.

[0008] The present invention also provides the use of the ginsenoside composition in the preparation of products that enhance the enzyme activity of alcohol dehydrogenase and / or acetaldehyde dehydrogenase or in the preparation of products that accelerate ethanol metabolism and / or acetaldehyde metabolism.

[0009] Preferably, the products that accelerate the metabolism of ethanol and / or acetaldehyde include products that have one or more functions such as protecting the liver, protecting the cardiovascular system, and relieving hangovers.

[0010] The present invention also provides a ginsenoside composition comprising the following components in parts by weight: 0.5-1 parts 20(S)-Rh1, 0.5-2.5 parts 20(R)-Rh1, 0.5-2 parts Rd, 0.5-1.8 parts F2, 0.5-3 parts Rg6, 0.5-4 parts Rg4, 0.5-10 parts Rk3, 0.5-15 parts Rh4, 0.5-10 parts 20(S)-Rg3, 0.5-8 parts 20(R)-Rg3, 0.5-20 parts Rk1, 0.5-25 parts Rg5 and 0.3-0.5 parts 20(S)-Rh2.

[0011] This invention also provides a method for preparing the ginsenoside composition described in the above technical solution, comprising the following steps: mixing ginseng ultrafine powder with methanol, first heating and extraction, cooling, and filtering to obtain a first filtrate and a first filter residue; mixing the first filter residue with an ethanol aqueous solution with a mass percentage of 50%, second heating and extraction, and filtering to obtain a second filtrate and a second filter residue; mixing the second filter residue with water, third heating and extraction, and filtering to obtain a third filtrate and a third filter residue; combining the first filtrate, second filtrate, and third filtrate, concentrating, drying, and pulverizing to obtain the ginsenoside composition.

[0012] Preferably, the temperature of the first heating extraction is 60~70℃, and the time of the first heating extraction is 0.5~1.5h; the temperature of the second heating extraction is 65~75℃, and the time of the second heating extraction is 0.5~1.5h; the temperature of the third heating extraction is 90~100℃, and the time of the third heating extraction is 0.5~1h.

[0013] This invention also provides the application of the ginsenoside composition or the ginsenoside extract described in the above-mentioned technical solutions in the preparation of products having any one or more of the effects described in ① to ⑤: ①Enhance the enzyme activity of alcohol dehydrogenase and / or acetaldehyde dehydrogenase; ②Accelerate ethanol metabolism and / or acetaldehyde metabolism; ③Reduce oxidative stress; ④ Restore mitochondrial function; ⑤ Protect the liver.

[0014] This invention provides a ginsenoside composition for accelerating acetaldehyde metabolism, comprising Rg5, Rk1, Rh4, Rk3, and 20(S)-Rg3. The ginsenoside composition of this invention can increase the enzyme activity of acetaldehyde dehydrogenase, accelerate acetaldehyde metabolism, and speed up the clearance of acetaldehyde from the human body.

[0015] Furthermore, based on the above-mentioned ginsenoside composition, the present invention adds three saponins: Rg6, Rg4, and 20(R)-Rg3. The addition of these three saponins enables the ginsenoside composition of the present invention to simultaneously enhance the enzyme activity of alcohol dehydrogenase and acetaldehyde dehydrogenase, and can be used to prepare products that accelerate ethanol metabolism and / or acetaldehyde metabolism.

[0016] Furthermore, this invention also provides a ginsenoside composition containing the above eight ginsenosides and 20(S)-Rh1, 20(R)-Rh1, Rd, F2, and 20(S)-Rh2, which can also be called Li-Ginseng Ginsenosides (LGG). Experimental verification shows that the ginsenoside composition of this invention can enhance the enzyme activity of alcohol dehydrogenase and / or acetaldehyde dehydrogenase, accelerate ethanol metabolism and / or acetaldehyde metabolism, reduce oxidative stress, and restore mitochondrial function, thereby protecting the liver. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 The LGG high performance liquid chromatography detection result diagram provided by the present invention; Figure 2 The MTT assay results of ethanol on L-02 human hepatocytes provided by this invention; Figure 3 The image shows the cytotoxic effect of LGG-protected ethanol on L-02 human hepatocytes provided by this invention. Figure 4 The effect of LGG on alcohol dehydrogenase and acetaldehyde dehydrogenase provided by this invention is shown in the figure. Figure 5The graph shows the effect of LGG on acetaldehyde content provided by this invention. Figure 6 The diagram showing the effect of LGG on ROS levels provided by this invention; Figure 7 The diagram shows the effect of LGG on mitochondrial membrane potential provided by this invention. Figure 8 The following graphs illustrate the effects of several major ginsenoside monomers and simple mixtures on alcohol dehydrogenase (ADH) activity, provided by this invention. A represents the results for a mixture of 0.25 μg / mL, 0.5 μg / mL, and 1 μg / mL equimolar ratios of Rg5+Rk1+Rh4+Rk3+20(S)-Rg3; B represents the results for a 0.25 μg / mL, 0.5 μg / mL, and 1 μg / mL equimolar ratio of 20(S)-Rg3 monomer; C represents the results for a 0.25 μg / mL, 0.5 μg / mL, and 1 μg / mL equimolar ratio of Rg5+Rk1 mixture; and D represents the results for a 0.25 μg / mL, 0.5 μg / mL, and 1 μg / mL equimolar ratio of Rh4+Rk3 mixture. Figure 9 The following graphs illustrate the effects of simple mixtures of several major ginsenosides on acetaldehyde dehydrogenase (ALDH) activity, provided by this invention. A represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg5+Rk1+Rh4+Rk3+20(S)-Rg3; B represents the results for a 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL 20(S)-Rg3 monomer; C represents the results for a 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratio of Rg5+Rk1; and D represents the results for a 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratio of Rh4+Rk3. Figure 10 The following graphs illustrate the effects of different ginsenosides on alcohol dehydrogenase (ADH) activity, as provided by this invention. A represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg6; B represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg4; C represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL 20(R)-Rg3; D represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg6+Rg4+20(R)-Rg3; and E represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg6+Rg4+20(R)-Rg3+Rg5+Rk1+Rh4+Rk3+20(S)-Rg3. Figure 11 The following graphs illustrate the effects of different ginsenosides on acetaldehyde dehydrogenase (ALDH) activity, as provided by this invention. A represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg6; B represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg4; C represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL 20(R)-Rg3; D represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg6+Rg4+20(R)-Rg3; and E represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg6+Rg4+20(R)-Rg3+Rg5+Rk1+Rh4+Rk3+20(S)-Rg3. Detailed Implementation

[0019] This invention provides a ginsenoside composition comprising the following components in parts by weight: 0.5-25 parts Rg5, 0.5-20 parts Rk1, 0.5-15 parts Rh4, 0.5-10 parts Rk3, and 0.5-10 parts 20(S)-Rg3. In a specific embodiment, the ginsenoside composition of this invention may consist of 0.5-25 parts Rg5, 0.5-20 parts Rk1, 0.5-15 parts Rh4, 0.5-10 parts Rk3, and 0.5-10 parts 20(S)-Rg3. In a specific embodiment, Rg5 in the ginsenoside composition may specifically be 1 part, 5 parts, 10 parts, 22.57 parts, or 25 parts. In a specific embodiment, Rk1 may specifically be 1 part, 5 parts, 10 parts, 16.12 parts, or 20 parts. In specific embodiments, Rh4 can be 1 part, 5 parts, 8 parts, 12.03 parts, or 15 parts. In specific embodiments, Rk3 can be 1 part, 3 parts, 5.65 parts, 8 parts, or 10 parts. In specific embodiments, 20(S)-Rg3 can be 1 part, 5 parts, 7.27 parts, or 10 parts. The ginsenoside composition of the present invention, composed of Rg5, Rk1, Rh4, Rk3, and 20(S)-Rg3, has been experimentally verified to accelerate acetaldehyde metabolism.

[0020] This invention also provides the application of the ginsenoside composition described in the above-mentioned technical solution in the preparation of products that enhance the enzyme activity of acetaldehyde dehydrogenase or in the preparation of products that accelerate acetaldehyde metabolism. In specific embodiments, the products that accelerate acetaldehyde metabolism include products having one or more functions such as protecting the liver, protecting the cardiovascular system, protecting the nervous system, and anti-inflammation. In this invention, the products also include excipients. In this invention, the excipients can be one or more of water, ethanol, cyclodextrin, starch, sucrose, and xylitol. In this invention, the products can be food, health food, and pharmaceuticals. The product limitations described below are the same as those described here and will not be repeated hereafter.

[0021] This invention also provides a ginsenoside composition comprising the following components in parts by weight: 0.5-3 parts Rg6, 0.5-4 parts Rg4, 0.5-8 parts 20(R)-Rg3, 0.5-25 parts Rg5, 0.5-20 parts Rk1, 0.5-15 parts Rh4, 0.5-10 parts Rk3, and 0.5-10 parts 20(S)-Rg3. In a specific embodiment, the ginsenoside composition may consist of 0.5-3 parts Rg6, 0.5-4 parts Rg4, 0.5-8 parts 20(R)-Rg3, 0.5-25 parts Rg5, 0.5-20 parts Rk1, 0.5-15 parts Rh4, 0.5-10 parts Rk3, and 0.5-10 parts 20(S)-Rg3. In a specific embodiment, Rg6 may be 1 part, 2.26 parts, or 3 parts. In specific embodiments, Rg4 can be 1 part, 2 parts, 3.72 parts, or 4 parts. In specific embodiments, 20(R)-Rg3 can be 1 part, 3 parts, 5 parts, or 7.94 parts. In specific embodiments, Rg5 in the ginsenoside composition can be 1 part, 5 parts, 10 parts, 22.57 parts, or 25 parts. In specific embodiments, Rk1 can be 1 part, 5 parts, 10 parts, 16.12 parts, or 20 parts. In specific embodiments, Rh4 can be 1 part, 5 parts, 8 parts, 12.03 parts, or 15 parts. In specific embodiments, Rk3 can be 1 part, 3 parts, 5.65 parts, 8 parts, or 10 parts. In specific embodiments, 20(S)-Rg3 can be 1 part, 5 parts, 7.27 parts, or 10 parts. Based on Rg5, Rk1, Rh4, Rk3 and 20(S)-Rg3, this invention adds three components, Rg6, Rg4 and 20(R)-Rg3, which enables the ginsenoside composition to not only improve the activity of acetaldehyde dehydrogenase, but also significantly improve the activity of alcohol dehydrogenase, thereby enabling it to be used to prepare products that accelerate ethanol metabolism and / or acetaldehyde metabolism.

[0022] This invention also provides the use of the ginsenoside composition in the preparation of products that enhance the activity of alcohol dehydrogenase and / or acetaldehyde dehydrogenase, or in the preparation of products that accelerate ethanol and / or acetaldehyde metabolism. In specific embodiments, the products that accelerate ethanol and / or acetaldehyde metabolism include products that have one or more functions such as protecting the liver, protecting the cardiovascular system, and alleviating hangovers.

[0023] This invention also provides a ginsenoside composition comprising the following components in parts by weight: 0.5-1 parts 20(S)-Rh1, 0.5-2.5 parts 20(R)-Rh1, 0.5-2 parts Rd, 0.5-1.8 parts F2, 0.5-3 parts Rg6, 0.5-4 parts Rg4, 0.5-10 parts Rk3, 0.5-15 parts Rh4, 0.5-10 parts 20(S)-Rg3, 0.5-8 parts 20(R)-Rg3, 0.5-20 parts Rk1, 0.5-25 parts Rg5, and 0.3-0.5 parts 20(S)-Rh2. In specific embodiments, Rg6 may be 1 part, 2.26 parts, or 3 parts. In specific embodiments, Rg4 may be 1 part, 2 parts, 3.72 parts, or 4 parts. In specific embodiments, the 20(R)-Rg3 can be 1 part, 3 parts, 5 parts, or 7.94 parts. In specific embodiments, the Rg5 in the ginsenoside composition can be 1 part, 5 parts, 10 parts, 22.57 parts, or 25 parts. In specific embodiments, the Rk1 can be 1 part, 5 parts, 10 parts, 16.12 parts, or 20 parts. In specific embodiments, the Rh4 can be 1 part, 5 parts, 8 parts, 12.03 parts, or 15 parts. In specific embodiments, the Rk3 can be 1 part, 3 parts, 5.65 parts, 8 parts, or 10 parts. In specific embodiments, the 20(S)-Rg3 can be 1 part, 5 parts, 7.27 parts, or 10 parts. In specific embodiments, the 20(S)-Rh1 can be 0.5 parts, 0.96 parts, or 1 part. In specific embodiments, the 20(R)-Rh1 can be 1 part, 2 parts, 2.47 parts, or 2.5 parts. In specific embodiments, the Rd can be 1 part, 1.75 parts, or 2 parts. In specific embodiments, the F2 can be 0.5 parts, 1 part, 1.52 parts, or 1.8 parts. In specific embodiments, the 20(S)-Rh2 can be 0.3 parts, 0.46 parts, or 0.5 parts. The ginsenoside composition of the present invention not only contains the above-mentioned 8 saponin components (Rg5, Rk1, Rh4, Rk3, 20(S)-Rg3, Rg6, Rg4, and 20(R)-Rg3), but also contains 20(S)-Rh1, 20(R)-Rh1, Rd, F2, and 20(S)-Rh2. The above-mentioned saponins work together to enhance the enzyme activity of alcohol dehydrogenase and / or aldehyde dehydrogenase; accelerate ethanol metabolism and / or aldehyde metabolism; reduce oxidative stress; restore mitochondrial function and protect the liver.

[0024] The present invention does not impose any particular limitation on the preparation method of the above-mentioned ginsenoside compositions. They can be obtained by combining conventional commercially available ginsenoside monomers or by preparation.

[0025] When the ginsenoside composition contains 0.5-1 parts of 20(S)-Rh1, 0.5-2.5 parts of 20(R)-Rh1, 0.5-2 parts of Rd, 0.5-1.8 parts of F2, 0.5-3 parts of Rg6, 0.5-4 parts of Rg4, 0.5-10 parts of Rk3, 0.5-15 parts of Rh4, 0.5-10 parts of 20(S)-Rg3, 0.5-8 parts of 20(R)-Rg3, 0.5-20 parts of Rk1, 0.5-25 parts of Rg5 and 0.3-0.5 parts of 20(S)-R At h2, the preparation method of the ginsenoside composition includes the following steps: mixing ginseng ultrafine powder with methanol, performing a first heating extraction, cooling, and filtering to obtain a first filtrate and a first filter residue; mixing the first filter residue with an ethanol aqueous solution with a mass percentage of 50%, performing a second heating extraction, and filtering to obtain a second filtrate and a second filter residue; mixing the second filter residue with water, performing a third heating extraction, and filtering to obtain a third filtrate and a third filter residue; combining the first filtrate, the second filtrate, and the third filtrate, concentrating, drying, and pulverizing to obtain the ginsenoside composition. In a specific embodiment, the temperature of the first heating extraction is 60~70℃, and the time of the first heating extraction is 0.5~1.5h, specifically, it can be heating extraction at 65℃ for 1h; the temperature of the second heating extraction is 65~75℃, and the time of the second heating extraction is 0.5~1.5h, specifically, it can be heating extraction at 70℃ for 1h; the temperature of the third heating extraction is 90~100℃, and the time of the third heating extraction is 0.5~1h, specifically, it can be heating extraction at 95℃ for 1h. In a specific embodiment, the ginseng ultrafine powder is a commercially available product, which can be purchased from Yanbian Andikanghua Biotechnology Co., Ltd. In a specific embodiment, the volume ratio of the ginseng ultrafine powder to methanol can be 1:(8~12), specifically 1:10. In a specific embodiment, the cooling is cooling to room temperature, which in this invention can be 20~30℃. In a specific embodiment, the volume ratio of the first filter residue to ethanol can be 1:(1.5~3), specifically 1:2. In a specific embodiment, the volume ratio of the second filter residue to water can be 1:(8~12), specifically 1:10. The ginsenoside composition product prepared by this invention using ginseng ultrafine powder can be called Li-Ginseng Ginsenosides (LGG).

[0026] This invention also provides the application of the ginsenoside composition or the ginsenoside extract described in the above-mentioned technical solutions in the preparation of products having any one or more of the effects described in ① to ⑤: ①Enhance the enzyme activity of alcohol dehydrogenase and / or acetaldehyde dehydrogenase; ②Accelerate ethanol metabolism and / or acetaldehyde metabolism; ③Reduce oxidative stress; ④ Restore mitochondrial function; ⑤ Protect the liver.

[0027] The ginsenoside composition (including LGG) of the present invention has the effects of enhancing the enzyme activity of alcohol dehydrogenase and / or acetaldehyde dehydrogenase; accelerating ethanol metabolism and / or acetaldehyde metabolism; reducing oxidative stress; restoring mitochondrial function; and protecting the liver.

[0028] To further illustrate the present invention, the following detailed description, in conjunction with embodiments, provides a ginsenoside composition for accelerating acetaldehyde metabolism, its preparation method, and its application. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0029] Example 1 Li-Ginseng Powder (LGP) was purchased from Yanbian Andikanghua Biotechnology Co., Ltd. Product Name: Li-Ginseng Powder (Solid Beverage); Food Safety License Number: SC10622240156068; Manufacturer: Yanbian Andikanghua Biotechnology Co., Ltd.; Production Address: No. 288, Changbai Road, Yanji City, Yanbian Korean Autonomous Prefecture, Jilin Province.

[0030] Li-Ginseng Ginsenosides (LGG) were extracted from ultrafine chestnut powder. The method was as follows: ultrafine chestnut powder was mixed with 10 times its volume of methanol, heated in a water bath (65℃) for 1 hour, cooled to room temperature (25℃), and filtered to obtain the first filtrate and the first filter residue. Then, 50% ethanol aqueous solution was added to the first filter residue at a volume ratio of 1:2, mixed evenly, heated in a water bath (70℃) for 1 hour, and filtered to obtain the second filtrate and the second filter residue. 10 times its volume of deionized water was added to the second filter residue, heated in a water bath at 95℃ for 1 hour, and filtered to obtain the third filtrate and the third filter residue. The first, second, and third filtrates were combined, concentrated using a rotary evaporator, and the concentrated solution was freeze-dried and ground into powder, which is LGG.

[0031] LGG was analyzed by high performance liquid chromatography.

[0032] High-performance liquid chromatography (HPLC) analysis was performed using a Waters 2695 HPLC system (Milford, MA) and a Waters 2489 UV-Vis detector system running Empower Pro (Waters) software. An Acchrom C-18 reversed-phase column with a particle size of 5 μm and an inner diameter of 4.6 × 250 mm was used. Experimental conditions were: column temperature 35℃, injection volume 50 μl, flow rate 1 mL / min, and detection wavelength 203 nm. The mobile phase was water (solvent A) and acetonitrile (solvent B). The HPLC solvent program was: 0 min, 20% (B), 80% (A); 20 min, 20% (B), 80% (A); 31 min, 32% (B), 68% (A); 40 min, 43% (B), 57% (A); 70 min, 100% (B), 0% (A), with gradient elution. The results are shown below. Figure 1 As shown in Table 1.

[0033] Table 1. Content of each saponin (percentage of total ginsenosides)

[0034] The above results indicate that the most common ginsenosides in ginseng (such as Rg1, Re and Rb1, which were not detected) have been converted into rare ginsenosides Rg6, Rg4, Rh4, Rk3, Rg3, Rg5, Rk1 and Rh2, which account for about 78% of the total ginsenoside content.

[0035] Example 2 Cell culture and cell experiments. L-02 human hepatocytes were cultured in 1640 medium containing 10% fetal bovine serum, streptomycin (100 μg / mL), and penicillin (100 μg / mL) at 37°C under humidified conditions of 5% carbon dioxide. The cytotoxicity of alcohol to L-02 human hepatocytes was determined using the MTT assay: L-02 human hepatocytes were seeded in 96-well plates (1 × 10⁶ cells / well). 4 L-02 human hepatocytes were cultured for 24 h with 100 μl of cell suspension. Then, 56% baijiu (Chinese white liquor) was serially diluted with 1640 medium to ethanol concentrations of 0 mmol / L, 100 mmol / L, 150 mmol / L, 200 mmol / L, 250 mmol / L, 300 mmol / L, 500 mmol / L, 800 mmol / L, and 1000 mmol / L, respectively, for 44 h. MTT (5 mg / mL, 20 μl per well) was then added and cultured for another 4 h. After discarding the culture medium, 150 μL of DMSO was added to each well, and the absorbance was measured at 550 nm. The results are as follows: Figure 2As shown. Similarly, the hepatoprotective effect of LGG against alcohol was determined. L-02 human hepatocytes were treated with diluted 56% baijiu (ethanol concentration 200 mmol / L) and the LGG of this invention (0 μg / mL, 0.5 μg / mL, and 1 μg / mL, dissolved in 200 mmol / L ethanol, respectively), while the control group was treated with 1640 medium. The results are shown in... Figure 3 As shown in the figure. Subsequent experiments all used the above concentration.

[0036] MTT assay results showed that ethanol inhibited the proliferation of L-02 human hepatocytes in a dose-dependent manner, with a half-maximal inhibitory concentration (IC50) of 1,500%. 50 Since the concentration of ethanol was 238 mM, a concentration of 200 mM ethanol was selected for subsequent experiments. Compared with the ethanol-treated group alone, LGG at concentrations of 0.5 µg / mL and 1 µg / mL significantly improved cell viability, indicating that LGG has a hepatoprotective effect.

[0037] Example 3 The effects of LGG on the enzyme activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), acetaldehyde content, reactive oxygen species (ROS), and mitochondrial membrane potential were analyzed at the cellular level. L-02 human hepatocytes were seeded in 6-well plates (3 × 10⁶ cells per well). 5 L-02 human hepatocytes were cultured for 24 h. Subsequently, the cells were treated with Erguotou liquor (ethanol concentration 200 mmol / L) and LGG (0 μg / mL, 0.5 μg / mL, 1 μg / mL) for 24 h, and then the cells were collected for analysis.

[0038] Alcohol dehydrogenase (ADH) Figure 4 A) and aldehyde dehydrogenase (ALDH) Figure 4 The enzyme activity assay results for B) are as follows: Figure 4 As shown in the figure. Enzyme activity assays revealed that ethanol exposure significantly reduced the activities of both ADH and ALDH enzymes compared to the untreated control group. However, compared to the ethanol-treated group alone, LGG significantly enhanced the activities of both ADH and ALDH enzymes, especially at a concentration of 1 µg / mL, indicating that it plays a positive role in promoting ethanol clearance.

[0039] Intracellular acetaldehyde content measurement results are as follows Figure 5 As shown in the figure, the acetaldehyde content results showed that the acetaldehyde concentration in the LGG-treated group was significantly lower than that in the ethanol-treated group alone, and even lower than that in the normal control group, indicating that LGG has a strong ability to remove acetaldehyde produced by ethanol metabolism and endogenous cellular processes.

[0040] Intracellular reactive oxygen species (ROS) level measurement results are as follows: Figure 6 As shown. The results of mitochondrial membrane potential measurement are as follows. Figure 7As shown ( Figure 7 The scale bar is 20 μm. Considering that ethanol metabolism also produces excessive reactive oxygen species (ROS), leading to oxidative stress and mitochondrial dysfunction, ROS and mitochondrial membrane potential were measured. The results showed that ethanol exposure led to a significant increase in total ROS levels and a significant decrease in mitochondrial membrane potential (compared to the control group), while LGG treatment effectively reversed these changes.

[0041] In summary, these results indicate that LGG exerts its hepatoprotective effect by enhancing the enzymatic activity of ADH and ALDH, promoting alcohol metabolism, acetaldehyde clearance, reducing oxidative stress, and restoring mitochondrial function.

[0042] Example 4 Based on the conclusions of Example 3, this example utilizes several specific major ginsenoside monomers / simple mixtures for cell experiments to detect their effects on the enzyme activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in L-02 human hepatocytes under ethanol-added / non-ethanol-added culture conditions. L-02 human hepatocytes were seeded in 6-well plates (3 × 10⁶ cells per well). 5 Cells were cultured for 24 h. The cultured cells were divided into two groups: a normal group (without ethanol) treated with different types and concentrations (0.25 μg / mL, 0.5 μg / mL, 1 μg / mL) of ginsenosides for 24 h; and an ethanol group treated with Erguotou liquor (200 mmol / L ethanol) and different types and concentrations (0.25 μg / mL, 0.5 μg / mL, 1 μg / mL) of ginsenosides for 24 h. Cells were collected for analysis. All ginsenoside monomers were purchased from Shanghai Yuanye Biotechnology Co., Ltd. Figure 8 Figures show the effects of several major ginsenoside monomers and simple mixtures on alcohol dehydrogenase (ADH) activity. A represents the results for a mixture of Rg5+Rk1+Rh4+Rk3+20(S)-Rg3 in equimolar ratios of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL; B represents the results for a 20(S)-Rg3 monomer in equimolar ratios of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL; C represents the results for a mixture of Rg5+Rk1 in equimolar ratios of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL; and D represents the results for a mixture of Rh4+Rk3 in equimolar ratios of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL. Figure 9The results show the effects of simple mixtures of several major ginsenosides on acetaldehyde dehydrogenase (ALDH) activity. A represents the results of mixtures with equimolar ratios of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg5+Rk1+Rh4+Rk3+20(S)-Rg3; B represents the results of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL 20(S)-Rg3 monomer; C represents the results of mixtures with equimolar ratios of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg5+Rk1; and D represents the results of mixtures with equimolar ratios of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rh4+Rk3. The results showed that, regardless of the presence or absence of exogenous ethanol, ginsenosides Rg5, Rk1, Rh4, Rk3, and 20(S)-Rg3 monomers and mixtures could not increase the activity of alcohol dehydrogenase (ADH) in human L-02 hepatocytes. Under normal culture conditions, ginsenosides Rg5, Rk1, Rh4, Rk3, and 20(S)-Rg3 monomers and mixtures could not increase the activity of aldehyde dehydrogenase (ALDH) in human L-02 hepatocytes. However, under the presence of exogenous ethanol, they could significantly increase the activity of ALDH in human L-02 hepatocytes, but not to the level comparable to LGG. Therefore, it can be concluded that other rare ginsenosides in LGG of this invention, such as ginsenosides Rg6, Rg4, and 20(R)-Rg3, play an important role in increasing the activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Since the raw material for LGG preparation, chestnut ginseng ultrafine powder, is a deep-processed product of ginseng as a whole, processed by bio-enzymatic conversion, the content of each component of the product is determined and the process is stable. The LGG prepared by this invention using chestnut ginseng ultrafine powder has a determined and stable content of ginsenoside components, which has a more prominent advantage in enhancing the enzyme activity of ADH and ALDH.

[0043] The results above indicate that mixtures of Rg5, Rk1, Rh4, Rk3 and 20(S)-Rg3, mixtures of Rg5 and Rk1, and mixtures of Rh4 and Rk3 did not increase the activity of alcohol dehydrogenase (ADH) in a dose-dependent manner. However, in the presence of ethanol, mixtures of the five saponins (Rg5, Rk1, Rh4, Rk3 and 20(S)-Rg3) significantly increased the activity of acetaldehyde dehydrogenase (ALDH) in a dose-dependent manner.

[0044] Example 5 Furthermore, this embodiment determined the effects of ginsenosides Rg6, Rg4, and 20(R)-Rg3 on the intracellular alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activities of L-02 human hepatocytes under ethanol-added and ethanol-free culture conditions. L-02 human hepatocytes were seeded in 6-well plates (3 × 10⁶ cells per well).5 Cells were cultured for 24 h. The cultured cells were divided into two groups: a normal group (without ethanol) treated with different types and concentrations (0.25 μg / mL, 0.5 μg / mL, 1 μg / mL) of ginsenosides for 24 h; and an ethanol group treated with Erguotou liquor (200 mmol / L ethanol) and different types and concentrations (0.25 μg / mL, 0.5 μg / mL, 1 μg / mL) of ginsenosides for 24 h. Cells were collected for analysis. All ginsenoside monomers were purchased from Shanghai Yuanye Biotechnology Co., Ltd. Figure 10 The results show the effects of different ginsenosides on alcohol dehydrogenase (ADH) activity. A represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg6; B represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg4; C represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL 20(R)-Rg3; D represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg6+Rg4+20(R)-Rg3; and E represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg6+Rg4+20(R)-Rg3+Rg5+Rk1+Rh4+Rk3+20(S)-Rg3. Figure 11 The results show the effects of different ginsenosides on acetaldehyde dehydrogenase (ALDH) activity. A represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg6; B represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL Rg4; C represents the results for 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL 20(R)-Rg3; D represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg6+Rg4+20(R)-Rg3; and E represents the results for a mixture of 0.25 μg / mL / 0.5 μg / mL / 1 μg / mL equimolar ratios of Rg6+Rg4+20(R)-Rg3+Rg5+Rk1+Rh4+Rk3+20(S)-Rg3.

[0045] The results showed that, regardless of the presence or absence of exogenous ethanol, ginsenosides Rg6, Rg4, and 20(R)-Rg3 monomers could not effectively increase the enzyme activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in L-02 human hepatocytes. When no exogenous ethanol was added to the system, the mixture of the three also failed to increase the enzyme activities of ADH and ALDH. However, when ethanol was present in the system, 1 A mixture of the three monomers (Rg6, Rg4, and 20(R)-Rg3) at a concentration of μg / mL can enhance the activity of ADH and ALDH. When ethanol is present in the system, the enzyme activity of ADH is significantly enhanced after treating L-02 human hepatocytes with a mixture of Rg6, Rg4, and 20(R)-Rg3 monomers with the other five major ginsenosides (Rg5, Rk1, Rh4, Rk3, and 20(S)-Rg3) of this invention. Regardless of whether exogenous ethanol is added, the activity of ALDH is increased in a dose-dependent manner after treating L-02 human hepatocytes with a mixture of Rg6, Rg4, and 20(R)-Rg3 monomers with the other five major ginsenosides (Rg5, Rk1, Rh4, Rk3, and 20(S)-Rg3) of this invention. The ALDH activity is significantly higher than that of the control group in the 0.5 μg / mL and 1 μg / mL groups without exogenous ethanol, as well as in the three groups with ethanol in the system. This indicates that the various saponins in this invention need to work synergistically to enhance the activity of ADH and ALDH, and Rg6, Rg4, and 20(R)-Rg3 are indispensable key components among them. At the same time, since the above eight saponin mixture can enhance the activity of ADH and ALDH in L-02 human hepatocytes without the addition of exogenous ethanol, it shows that under normal physiological conditions, the above eight saponin composition can still enhance the acetaldehyde metabolism rate in the human body.

[0046] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A ginsenoside composition, characterized in that, It includes the following components in parts by weight: 0.5 to 25 parts Rg5, 0.5 to 20 parts Rk1, 0.5 to 15 parts Rh4, 0.5 to 10 parts Rk3 and 0.5 to 10 parts 20(S)-Rg3.

2. The use of the ginsenoside composition of claim 1 in the preparation of products that enhance the enzyme activity of acetaldehyde dehydrogenase or in the preparation of products that accelerate acetaldehyde metabolism.

3. The application according to claim 2, characterized in that, The products that accelerate acetaldehyde metabolism include those with one or more functions such as protecting the liver, protecting the cardiovascular system, protecting the nervous system, and anti-inflammation.

4. A ginsenoside composition, characterized in that, It includes the following components in parts by weight: 0.5 to 3 parts Rg6, 0.5 to 4 parts Rg4, 0.5 to 8 parts 20(R)-Rg3, 0.5 to 25 parts Rg5, 0.5 to 20 parts Rk1, 0.5 to 15 parts Rh4, 0.5 to 10 parts Rk3 and 0.5 to 10 parts 20(S)-Rg3.

5. The use of the ginsenoside composition of claim 4 in the preparation of products that enhance the enzyme activity of alcohol dehydrogenase and / or acetaldehyde dehydrogenase or in the preparation of products that accelerate ethanol metabolism and / or acetaldehyde metabolism.

6. The application according to claim 5, characterized in that, The products that accelerate the metabolism of ethanol and / or acetaldehyde include those that have one or more functions such as protecting the liver, protecting the cardiovascular system, and relieving hangovers.

7. A ginsenoside composition, characterized in that, It includes the following components in parts by weight: 0.5 to 1 part 20(S)-Rh1, 0.5 to 2.5 parts 20(R)-Rh1, 0.5 to 2 parts Rd, 0.5 to 1.8 parts F2, 0.5 to 3 parts Rg6, 0.5 to 4 parts Rg4, 0.5 to 10 parts Rk3, 0.5 to 15 parts Rh4, 0.5 to 10 parts 20(S)-Rg3, 0.5 to 8 parts 20(R)-Rg3, 0.5 to 20 parts Rk1, 0.5 to 25 parts Rg5 and 0.3 to 0.5 parts 20(S)-Rh2.

8. The method for preparing the ginsenoside composition according to claim 7, characterized in that, Includes the following steps: Ginseng ultrafine powder was mixed with methanol, subjected to a first heating extraction, cooled, and filtered to obtain a first filtrate and a first filter residue. The first filter residue was mixed with a 50% (w / w) ethanol aqueous solution, subjected to a second heating extraction, and filtered to obtain a second filtrate and a second filter residue. The second filter residue was mixed with water, subjected to a third heating extraction, and filtered to obtain a third filtrate and a third filter residue. The first, second, and third filtrates were combined, concentrated, dried, and pulverized to obtain a ginsenoside composition.

9. The preparation method according to claim 8, characterized in that, The first heating extraction temperature is 60~70℃, and the first heating extraction time is 0.5~1.5h; the second heating extraction temperature is 65~75℃, and the second heating extraction time is 0.5~1.5h; the third heating extraction temperature is 90~100℃, and the third heating extraction time is 0.5~1h.

10. The use of the ginsenoside composition of claim 7 or the ginsenoside extract of claim 8 or 9 in the preparation of products having any one or more of the effects described in ① to ⑤: ①Enhance the enzyme activity of alcohol dehydrogenase and / or acetaldehyde dehydrogenase; ②Accelerate ethanol metabolism and / or acetaldehyde metabolism; ③Reduce oxidative stress; ④ Restore mitochondrial function; ⑤ Protect the liver.

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