Use of gaoyao-4 powder in preparation of a medicine for treating or preventing acute alcoholic liver

Abstract

The application provides application of Gao Yao-4 medicine powder in preparation of a medicine for treating or preventing acute alcoholic liver, and belongs to the field of medicine. It is found for the first time that the drugs in the Gao Yao-4 medicine powder have a combined effect, can effectively reduce the content of AST, ALT and TBIL in serum and liver tissue after a large amount of drinking, effectively reduce the content of TC, TG, MDA, TNF-alpha, IL-1beta and IL-6 in liver tissue after a large amount of drinking, improve the content of SOD, reduce the apoptosis rate of liver tissue cells, improve the expression amount of Bc1-2 protein, and reduce the expression amount of Bax, caspase3 and C-caspase3 protein, and has a repairing and protecting effect on alcoholic liver cell damage. It is shown that the Gao Yao-4 medicine powder can be used for treating / preventing acute alcoholic liver damage, and does not have side effects, and has high safety.

Description

Technical Field

[0001] This invention belongs to the field of medicine, specifically relating to the application of Gaoyao-4-Flavor Powder in the preparation of drugs for the treatment or prevention of acute alcoholic liver disease. Background Technology

[0002] Acute alcoholic liver injury refers to damage to hepatocytes and liver lobules caused by excessive alcohol consumption within a short period. It is an alcoholic viral hepatitis and liver injury resulting from alcohol poisoning and is a major cause of liver damage. Symptoms of acute alcoholic liver injury include alcoholic fatty liver, alcoholic hepatitis, and alcoholic cirrhosis. The goal of treatment for acute alcoholic liver injury is to delay and reduce the occurrence of liver failure, decompensated cirrhosis, liver cancer, and other related complications as much as possible for long-term alcohol drinkers, thereby improving their quality of life and prolonging their survival time.

[0003] While modern medicine has made significant progress in the diagnosis and treatment of acute alcoholic liver disease, several key challenges remain regarding long-term efficacy, such as drug side effects, recurrence rates, and the cost of long-term treatment. This is particularly true for decompensated cirrhosis, where there are no specific drugs, and for liver cancer, issues such as adverse reactions and recurrence caused by overtreatment. Therefore, addressing these challenges requires a comprehensive treatment approach involving multiple disciplines, modalities, and medications.

[0004] "Gaoyao-4-Flavor Powder" is a specialty Mongolian medicine from Alashan, also known as "Zhuangxi-4-Flavor Powder" or "Gaoyou-4-Flavor Powder." It consists of four herbs: Cynomorium songaricum, calcined gypsum, licorice root, and rock sugar. Appearance: Light brownish-white powder; taste: sweet and slightly bitter. Functions and indications: Relieves stagnation, aids digestion, stops vomiting, and stops diarrhea. Used for indigestion, acid reflux, bloating, headache, diarrhea, and fever in women. Currently, there are no reports of using "Gaoyao-4-Flavor Powder" for the treatment / prevention of acute alcoholic liver injury. Summary of the Invention

[0005] Therefore, the purpose of this invention is to provide the application of Gaoyao-4wei powder in the preparation of drugs for treating or preventing acute alcoholic liver disease. Gaoyao-4wei powder can be used to treat / prevent acute alcoholic liver injury and has no side effects, thus having high safety.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides the application of Gaoyao-4-flavor powder in the preparation of medicaments for the treatment or prevention of acute alcoholic liver disease.

[0008] Preferably, the Gaoyao-4wei powder is one or the only effective ingredient of the drug for treating or preventing alcoholic liver injury.

[0009] Preferably, the Gaoyao-4 Flavor Powder is made from 8g of Cynomorium songaricum, 7g of calcined gypsum, 7g of sclerotium buds, and 2.5g of rock sugar by weight.

[0010] Preferably, the effective dosage of the Gaoyao-4 Flavor Powder is 3-9 g / day for adults.

[0011] Preferably, the Gaoyao-4 Flavor Powder repairs and / or protects against alcoholic hepatocellular damage.

[0012] The present invention also provides a medicament for treating or preventing acute alcoholic liver disease, comprising an effective dose of Gaoyao-4wei powder.

[0013] Preferably, powders, decoctions, pills, capsules, tablets, granules, and oral liquids are prepared by adding pharmaceutically acceptable excipients.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] This invention reveals that the various herbs in Gaoyao-4-Flavor Powder have a synergistic effect in treating acute alcoholic liver disease. They can effectively reduce the levels of AST, ALT, and TBIL in serum and liver tissue after heavy alcohol consumption, effectively reduce the levels of TC, TG, MDA, TNF-α, IL-1β, and IL-6 in liver tissue after heavy alcohol consumption, increase SOD levels, reduce liver cell apoptosis rate, increase Bcl-2 protein expression, and reduce Bax, caspase3, and C-caspase3 protein expression, thus exhibiting repair and protective effects against alcoholic hepatocellular damage. This indicates that Gaoyao-4-Flavor Powder can be used to treat / prevent acute alcoholic liver injury without side effects and has high safety. Attached Figure Description

[0016] Figure 1 The results of liver tissue HE pathological sections are as follows: A is the blank group, B is the model group, C is the Western medicine group, D is the high-dose Mongolian medicine group, E is the medium-dose Mongolian medicine group, and F is the low-dose Mongolian medicine group. Detailed Implementation

[0017] This invention provides the application of Gaoyao-4wei powder in the preparation of a drug for treating or preventing acute alcoholic liver injury, wherein Gaoyao-4wei powder is one or the only effective ingredient of the drug for treating or preventing alcoholic liver injury, and the effective dose of Gaoyao-4wei powder is 3-9 g / day for adults.

[0018] The Gaoyao-4 Flavor Powder of the present invention repairs and / or protects alcoholic hepatocyte injury, can effectively reduce the contents of AST, ALT, and TBIL in serum and liver tissue after a large amount of alcohol consumption, effectively reduce the contents of TC, TG, MDA, TNF-α, IL-1β, and IL-6 in liver tissue after a large amount of alcohol consumption, increase the content of SOD, reduce the apoptosis rate of liver tissue cells, increase the expression level of Bc1-2 protein, and reduce the expression levels of Bax, caspase3, and C-caspase3 proteins.

[0019] The Gaoyao-4 Flavor Powder of the present invention can be used orally. The Gaoyao-4 Flavor Powder is prepared from 8 g of Cynomorium songaricum Rupr., 7 g of calcined Gypsum Fibrosum, 7 g of Gentianopsis barbata (Froel.) Ma, and 2.5 g of rock sugar by mass parts.

[0020] The present invention also provides a drug for treating or preventing acute alcoholic liver, which includes an effective dose of Gaoyao-4 Flavor Powder. The drug of the present invention can take Gaoyao-4 Flavor Powder as the only active ingredient, or can use Gaoyao-4 Flavor Powder and other active ingredients with liver protection or anti-alcohol effects together as active ingredients.

[0021] The drug of the present invention also includes pharmaceutically acceptable excipients, and different dosage forms are prepared by adding excipients. The dosage forms include powder, decoction, pill, capsule, tablet, granule, and oral liquid.

[0022] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they cannot be understood as limiting the protection scope of the present invention.

[0023] In the specific embodiments of the present invention, the experimental animals are C57BL / 6 mice (SPF grade), male, with a body weight of 18-22 g, purchased from Beijing Speyford Biotechnology Co., Ltd., license number SCXK (Beijing) 2019-0010, and raised under conventional conditions; Bifendate Pills (batch number: A02J191109), purchased from Wanbond Pharmaceutical Group Co., Ltd.; Lieber-DeCarli alcohol liquid diet (code: TP4030D) and Lieber-DeCarli control liquid diet (code: TP4030C), purchased from Nantong Trofi Feed Technology Co., Ltd.

[0024] In the specific embodiments of the present invention, the TBIL kit, ALT kit, AST kit, TG kit, and TC kit are purchased from Changchun Huili Biotechnology Co., Ltd.; the TNF-α ELISA kit, IL-6 ELISA kit, and IL-1β ELISA kit are purchased from Thermo Fisher Biotechnology Co., Ltd.; the SOD test kit and MDA test kit are purchased from Nanjing Jiancheng Co., Ltd.

[0025] In a specific embodiment of this invention, the Mongolian medicine "Gaoyao-4-Flavor Powder" is prepared by pulverizing Cynomorium songaricum, calcined gypsum, angelica dahurica, and rock sugar separately, and then mixing them in a certain proportion (prepared by the prescription dispensing room of the Traditional Chinese and Mongolian Medicine Hospital of Alxa Left Banner, Inner Mongolia). The prepared Mongolian medicine "Gaoyao-4-Flavor Powder" is taken with purified water.

[0026] In a specific embodiment of the present invention, statistical processing was performed using SPSS 24.00 software.

[0027] Calculation analysis, experimental data The comparison between groups was performed using one-way ANOVA, and P < 0.05 was considered statistically significant.

[0028] Example 1

[0029] Acute toxicity test of Mongolian medicine "Gaoyao-4-Flavor Powder":

[0030] Twenty-four healthy mice of similar body weight, half male and half female, were randomly divided into four groups: a control group, a low-dose group, a medium-dose group, and a high-dose group. Each group consisted of six mice, half male and half female. The dosages for the low-dose, medium-dose, and high-dose groups were set at 0.6 g / kg, 1.2 g / kg, and 2.4 g / kg, respectively. The maximum tolerable gavage volume for mice was 40 mL / kg, and the mice were administered the medication once daily via gavage.

[0031] Experimental Results: In the treatment group, 24 mice showed reduced activity and abdominal distension 4-5 minutes after gavage administration. These symptoms disappeared after 24 hours. Their fur remained shiny, and no abnormal secretions were found in the nose, eyes, or mouth. No abnormalities were observed in activity, diet, or feces, and no deaths occurred. Post-euthanasia, gross examination of major organs revealed no significant abnormalities. Compared to the control group, no significant abnormalities were observed. This indicates that the test drug had no significant effect on the general condition and organs of the mice. The control group mice showed good mental state, shiny white fur, normal urination and defecation, and no deaths. This indicates that Gaoyao-4wei San has low toxicity and is safe and reliable for clinical application.

[0032] Example 2

[0033] The protective effect of Mongolian medicine "Gaoyao-4-Flavor Powder" against alcoholic liver injury induced by liquid alcohol feed in mice:

[0034] 1. Animal grouping and administration

[0035] Forty-two mice were randomly divided into six groups: a blank control group, a model group, a Western medicine group, and low-, medium-, and high-dose Mongolian medicine groups, with seven mice in each group. The Western medicine group was given biphenyl diester at 2.9 mg / kg; the Mongolian medicine groups were given "Gaoyao-4-Flavor Powder" at 0.6 g / kg, 1.2 g / kg, and 2.4 g / kg, respectively. Administered at fixed times (7-9 AM / day), once daily, for 10 consecutive days.

[0036] 2. Model Establishment

[0037] Mice were acclimatized for one week, then given a standard Lieber-DeCarli liquid diet (Lieber-DeCarli control liquid diet) for 5 days to allow for acclimatization. The control group continued to receive the standard LD liquid diet, while the other five groups received a 5% ethanol LD liquid diet (Lieber-DeCarli alcohol liquid diet) for 10 consecutive days. During the modeling process, the Western medicine group and the low, medium, and high dose Mongolian medicine groups were administered the drugs, while the control and model groups received an equal volume of physiological saline. One hour after the last administration, all groups except the control group were given a single dose of 0.066 L / kg of high-concentration baijiu (31.5% alcohol solution) to induce an acute liver injury model.

[0038] 3. Detection indicators

[0039] (1) Statistical analysis of the changes in body weight of mice in each group

[0040] Table 1 Comparison of body weight changes in mice of different groups

[0041]

[0042] Note: Compared with the control group, #P<0.05, ##P<0.01; compared with the model group, *P<0.05, **P<0.01.

[0043] As shown in Table 1, compared with the control group, the body weight of the model group did not differ significantly in the early stage, but showed a significant difference after modeling with alcohol-based liquid feed (P<0.05, P<0.01). Compared with the model group, the body weight of the high-, medium-dose Mongolian medicine group and the Western medicine group did not differ significantly in the early stage, but increased after modeling with alcohol-based liquid feed (P<0.05, P<0.01). The low-dose group showed no significant difference from the model group.

[0044] (2) Measurement of serum AST, ALT, and TBIL levels

[0045] One hour after the last gavage administration, and after a 24-hour period of fasting with free access to water, blood was collected from the orbital cavity. After clotting, the blood was centrifuged at 3000 rpm for 10 minutes to separate the serum. The supernatant was then used to detect the AST, ALT, and TBIL levels according to the kit instructions. The results are shown in Table 2.

[0046] Table 2 Comparison of serum AST, ALT, and TBIL levels in mice of different groups

[0047]

[0048]

[0049] Note: Compared with the control group, #P<0.05, ##P<0.01; compared with the model group, *P<0.05, **P<0.01.

[0050] As shown in Table 2, compared with the control group, the serum AST, ALT, and TBIL levels in the model group mice were significantly increased, with statistically significant differences (P<0.05), indicating successful modeling. Compared with the model group, the serum AST, ALT, and TBIL levels in the high- and medium-dose Mongolian medicine groups and the Western medicine group were significantly decreased, with statistically significant differences (P<0.05, P<0.01), indicating that the high- and medium-dose Mongolian medicine "Gaoyao-4wei San" can effectively reduce the serum AST, ALT, and TBIL levels after excessive alcohol consumption.

[0051] (3) Measurement of TG, TC, SOD, MDA, TNF-α, and IL-6 levels in liver tissue

[0052] Liver tissues from mice in each group were pre-cooled at 4℃, washed with physiological saline, and homogenized to a 10% concentration. The homogenate was centrifuged at 3000 rpm for 10 min at 4℃. The supernatant was collected, and the levels of TNF-α, IL-6, IL-1β, TG, TC, SOD, and MDA were measured using TNF-α ELISA, IL-6 ELISA, IL-1β ELISA, TG, TC, and SOD according to the kit instructions. The results are shown in Tables 3 and 4.

[0053] Table 3 Comparison of TNF-α, IL-1β, and IL-6 levels in liver tissue of mice in each group

[0054]

[0055] Table 4 Comparison of TG, TC, SOD, and MDA contents in liver tissue of mice in each group

[0056]

[0057]

[0058] Note: Compared with the control group, #P<0.05, ##P<0.01; compared with the model group, *P<0.05, **P<0.01.

[0059] The results showed that, compared with the control group, the levels of TNF-α, IL-1β, and IL-6 in the liver tissue of mice in the model group were significantly increased (P<0.05), indicating successful modeling. Compared with the model group, the levels of TNF-α, IL-1β, and IL-6 in the liver tissue of the high-, medium-, and low-dose Mongolian medicine groups and the Western medicine group were significantly decreased (P<0.05, P<0.01), with no significant difference in TNF-α content in the liver tissue of the low-dose group. This indicates that the high- and medium-dose groups of the Mongolian medicine "Gaoyao-4wei San" can effectively reduce the levels of TNF-α, IL-1β, and IL-6 in the liver tissue after heavy alcohol consumption.

[0060] Compared with the control group, the levels of MDA, TG, and TC in the liver tissue of mice in the model group were significantly increased, while the level of SOD was significantly decreased, with statistically significant differences (P<0.05), indicating successful modeling. Compared with the model group, the levels of TC and TG in the liver tissue of the high-, medium-, and low-dose Mongolian medicine groups and the Western medicine group decreased, with statistically significant differences (P<0.05); the levels of MDA in the liver tissue of the high- and medium-dose Mongolian medicine groups and the Western medicine group decreased, while the levels of SOD increased, with statistically significant differences (P<0.05), with no significant difference in SOD and MDA levels in the low-dose group. This indicates that the high- and medium-dose groups of the Mongolian medicine "Gaoyao-4-Flavor Powder" can effectively reduce the levels of TC, TG, and MDA in the liver tissue and increase the level of SOD after heavy alcohol consumption.

[0061] (4) Observation of liver pathological morphology

[0062] Tissue samples were harvested from the same location in the livers of mice in each group, fixed in 4% paraformaldehyde, dehydrated, embedded in paraffin, stained with hematoxylin and eosin, sectioned, air-dried, and observed under a light microscope for morphological changes. Results are as follows: Figure 1 As shown, Figure 1 In the study, A was the blank group, B was the model group, C was the Western medicine group, D was the high-dose Mongolian medicine group, E was the medium-dose Mongolian medicine group, and F was the low-dose Mongolian medicine group.

[0063] HE pathological section analysis of liver tissue showed that the liver tissue structure of mice in the blank control group was normal, with hepatocytes arranged neatly and no pathological changes such as hepatocyte degeneration or necrosis were observed. In the model group, the liver tissue structure and hepatocyte arrangement were disordered, with most hepatocytes showing edema and fat droplets of varying sizes present in the liver tissue. The liver tissue structure of mice in the high-dose Mongolian medicine group and the Western medicine group was clearer, with hepatocytes arranged neatly and uniformly in size, and the fat droplet vacuoles were significantly reduced compared to the model group. In the medium-dose Mongolian medicine group, the liver tissue structure and hepatocyte arrangement were relatively neat, and hepatocyte edema and fat droplet vacuoles were reduced compared to the model group. The pathological morphology of the liver tissue in the low-dose Mongolian medicine group showed no significant improvement. This indicates that high and medium doses of the Mongolian medicine "Gaoyao-4-Flavor Powder" have a repairing and protective effect on alcoholic hepatocellular damage.

[0064] (5) Comparison of hepatocyte apoptosis in liver tissue

[0065] Liver tissues were collected from each group of mice, and the apoptosis of hepatocytes in the liver tissues of each group of mice was detected. The results are shown in Table 5.

[0066] Table 5 Comparison of apoptosis in liver tissue of mice in each group

[0067]

[0068] Note: Compared with the control group, #P<0.05, ##P<0.01; compared with the model group, *P<0.05, **P<0.01.

[0069] As shown in Table 5, compared with the blank group, the apoptosis rate of liver tissue cells in the model group was significantly increased, and the difference was statistically significant (P<0.01). Compared with the model group, the apoptosis rate of cells in the high, medium and low dose groups of Mongolian medicine was decreased, and the differences were statistically significant (P<0.05), indicating that the anti-apoptotic effect was positively correlated with the dose.

[0070] (6) Comparison of protein expression levels of Bcl-2, Bax, caspase3, and C-caspase3 in liver tissue

[0071] Liver tissues were collected from mice in each group, and the expression levels of Bc1-2, Bax, caspase3, and C-caspase3 proteins in the liver tissues of mice in each group were detected. The results are shown in Table 6.

[0072] Table 6 Comparison of protein expression levels in liver tissue of mice in each group

[0073]

[0074]

[0075] Note: Compared with the control group, #P<0.05, ##P<0.01; compared with the model group, *P<0.05, **P<0.01.

[0076] As shown in Table 6, compared with the control group, the Bcl-2 level in the liver tissue of mice in the model group was significantly decreased, while Bax, caspase 3, and C-caspase 3 were significantly increased, with statistically significant differences (P<0.05). Compared with the model group, Bcl-2 was significantly increased in the high- and medium-dose Mongolian medicine groups and the Western medicine group, with statistically significant differences (P<0.01), while there was no significant difference in Bcl-2 between the low-dose group and the model group. Compared with the model group, Bax, caspase 3, and C-caspase 3 were significantly decreased in the high-, medium-, and low-dose Mongolian medicine groups and the Western medicine group, with statistically significant differences (P<0.01). The anti-apoptotic effect was positively correlated with the dosage.

[0077] In summary, the high and medium dose groups of the Mongolian medicine "Gaoyao-4-Flavor Powder" showed varying degrees of improvement in liver function indicators and inhibited hepatocyte apoptosis and oxidative stress response, indicating that the Mongolian medicine "Gaoyao-4-Flavor Powder" has liver-protective and anti-liver-damage effects.

[0078] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. The application of Gaoyao-4-Flavor Powder as the sole effective ingredient in the preparation of drugs for the treatment or prevention of acute alcoholic liver injury, characterized in that... The Gaoyao-4 Flavor Powder, by weight, is made from 8g of Cynomorium songaricum, 7g of calcined gypsum, 7g of sclerotium buds, and 2.5g of rock sugar.

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