Traditional Chinese medicine composition for treating alcoholic liver injury, preparation and application thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine, and discloses a traditional Chinese medicine composition for treating alcoholic liver injury, a preparation and application thereof, the optimal medicine ratio is determined, the traditional Chinese medicine composition is dendrobium officinale liver protection soup, and the active ingredient raw materials of the traditional Chinese medicine composition comprise the following raw materials in parts by weight: 5-15 parts of dendrobium officinale, 6-10 parts of turmeric, 10-20 parts of radix puerariae, 6-12 parts of medicated leaven and 2-6 parts of amomum cardamomum. The dendrobium huoshanense liver protection decoction can remarkably reduce the levels of ALT, AST and TG in mouse serum, reduce the content of MDA in the liver and the levels of IL-1beta, IL-6 and TNF-alpha, improve the SOD level, relieve fat deposition in the mouse liver, relieve pathological damage to the mouse liver and relieve the oxidative stress level and the inflammatory factor level, and has a good protection effect on the liver with alcoholic liver damage.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and specifically, to a traditional Chinese medicine composition, preparation and application for treating alcoholic liver injury. Background Art

[0003] Acute alcohol-induced liver injury (AALI) refers to a series of pathological changes such as disorder of liver tissue structure and damage of liver cells induced by excessive alcohol intake in a short period, exceeding the metabolic capacity of the liver, resulting in abnormal liver function, and at the same time triggering oxidative stress and inflammatory responses in the body. Long-term alcohol consumption leads to the development of multiple stages such as alcoholic fatty liver (AFL), alcoholic hepatitis, liver fibrosis, and liver cancer. Its pathogenesis is related to multiple aspects such as enzyme activity changes caused by ethanol, oxidative stress, mitochondrial dysfunction, cytokine-mediated inflammation, disruption of the intestinal barrier, and intestinal dysbiosis.

[0004] In terms of treatment, reducing the blood concentration of alcohol, accelerating the metabolism of alcohol in the body, relieving the symptoms of drunkenness, and reducing the damage of alcohol to organs such as the liver and cardiovascular system by ingesting functional products are the current main anti-alcoholism mechanisms. The treatment guidelines for ALD in China propose that patients need to be given nutritional support on the basis of abstaining from alcohol to reduce the continuous stimulation of alcohol and improve the malnutrition caused by alcohol intake. The drugs mainly include silymarin, bicyclol, reduced glutathione (GSH), glycyrrhizin preparations, glucocorticoids, S-adenosylmethionine, polyene phosphatidylcholine, etc., which can improve biochemical indexes such as aspartate aminotransferase (AST) and alanine aminotransferase (ALT) of AFL; metadoxine can promote the clearance of alcohol in the blood and improve alcohol poisoning. Although the above drugs are used to treat ALD, their curative effects are limited. The effect of silymarin is slow, and one course of treatment is 3 months; reduced glutathione is often used in combination with other liver-protecting drugs as an adjuvant; the 28-day survival rate of patients with severe alcoholic hepatitis can be effectively improved after using glucocorticoids, but this drug has no effect on the survival rate at 3 months and 6 months. In addition, adverse reactions may also occur during use. Silymarin can form a complex with ferric ions, causing iron-deficiency anemia, and long-term use of glucocorticoids can increase the risk of infection. Therefore, finding drugs with good anti-alcoholism and liver-protecting activities and small toxic and side effects is still an urgent need in clinical treatment of patients with acute alcoholic liver injury, and it has very profound significance for the prevention of ALD and the truncation of the later disease development.

[0005] In traditional Chinese medicine (TCM), alcoholic liver disease falls within the categories of "injured by alcohol", "aversion to alcohol", "alcohol-induced derangement", "drunkenness", "alcohol syncope", "alcoholic jaundice", "alcoholic mass", "alcoholic tympanites", "alcoholic accumulation", and "alcoholic disease". "Injured by alcohol", "aversion to alcohol", "alcohol-induced derangement", "drunkenness", and "alcohol syncope" should be manifestations of acute alcoholic liver disease, while "alcoholic jaundice", "alcoholic mass", "alcoholic tympanites", and "alcoholic accumulation" represent various stages of alcoholic fatty liver, liver fibrosis, liver cirrhosis, and liver cancer. "Alcoholic disease" is the general term. "Treatise on the Causes and Symptoms of Various Diseases" states: "Alcohol has a pungent and fierce qi and is highly toxic. The liver is floating and the gallbladder is transverse, leading to wild and perverse anger, so it is called aversion to alcohol." Zhao Ji of the Song Dynasty recorded in "Complete Compendium of Holy Relief, Volume 73": "For those with weak stomachs, due to excessive drinking, the nature of alcohol is pungent and hot, prone to thirst and excessive drinking. When encountering qi obstruction in the air passage, both alcohol and the drink cannot be digested and accumulate in the hypochondrium, forming a mass." Therefore, for alcoholic disease, excessive drinking without restraint is the external cause, and congenital endowment deficiency and weak spleen and stomach are the internal causes. The formation of alcoholic disease is closely related to qi, blood, and body fluids. "Huangdi Neijing" says: "Alcohol is the essence of water and grains and the liquid of cooked grains. Its qi is pungent and fierce. When it enters the stomach, the stomach swells, qi rebels upward, fills the chest, and the liver is floating and the gallbladder is transverse." Alcohol is a product of dampness and heat and is toxic. It is warm in nature, sweet, pungent, and dispersing, entering the heart, lungs, liver, and stomach meridians, and is prone to generating dampness and heat. If one drinks excessively, it can disrupt qi and blood, causing disharmony between yin and yang. "Preserving Life with Supreme Secrets" also has the saying that "drinking in the morning injures the stomach, and residual alcohol injures the spleen". Excessive drinking without restraint is the fundamental cause of this disease, and congenital endowment deficiency or weak spleen and stomach is the internal factor. Alcohol heat injures body fluids, alcohol dampness stagnates the spleen and stomach, and alcohol toxin disrupts liver qi. The combination of these three leads to alcoholic disease.

[0006] The view in Li Dongyuan's "Treatise on the Spleen and Stomach" in the Jin Dynasty, "Alcohol is extremely hot and toxic, with both its qi and flavor being yang and it being an intangible substance. If injured by it, one should only induce sweating, and recovery will occur after sweating. Secondly, there is nothing better than promoting urination. These two methods can dispel dampness from the upper and lower parts respectively.", has been widely spread and is considered the main principle for alcoholic disease. Most of the ancient anti-alcoholic prescriptions also considered inducing sweating and promoting diuresis, inducing vomiting of alcohol and food, purging and reducing accumulation, and clearing heat and promoting diuresis. The inventor summarized clinical experience and believes that the nature of alcohol is pungent and hot, entering the collaterals and blood, prone to injuring body fluids, damaging the spleen and stomach, and injuring the true yin. Alcohol toxin and dampness stagnation further obstruct qi movement, and qi movement and collaterals dredging are required.

[0007] Modern research believes that the causes of alcoholic fatty liver are closely related to the indirect injury of the liver through the enterohepatic interaction, the direct injury of the liver by alcohol metabolism, hypoxia, etc. As an important immune organ of the body, the liver controls all types of immune cell populations and has the ability to recruit and activate related immune cells to respond to enterogenic metabolic and pathogenic signals. Alcohol changes the composition of gut microbiota, weakens the integrity and barrier function of the gut, and exacerbates enterogenic endotoxemia, which is the first and most important step in the occurrence of ALD.

[0008] Alcohol and its metabolite acetaldehyde generate reactive oxygen species (ROS) and reactive nitrogen species during liver metabolism, causing oxidative stress (OS), mitochondrial dysfunction, and ultimately leading to inflammation and even apoptosis. Hypoxia can cause necrosis of cells in the centrilobular region. Tissue hypoxia increases the level of the transcription factor HIF in hepatocytes by reducing the degradation of HIF in the proteasome. Many genes involved in liver inflammation, steatosis, fibrosis, angiogenesis, and tumorigenesis are regulated by HIF, accelerating the progression of ALD. Current research has found that mitochondrial metabolism and ROS are crucial for the regulation of HIFs, and HIFs have also been shown to regulate mitochondrial metabolism and ROS levels, and can regulate lipid metabolism in alcoholic fatty liver disease through mitophagy.

[0009] The inventors analyzed and found that Huhuangan Liver Protecting Decoction improves alcoholic liver disease through the pi3k-akt and HIF-1 signaling pathways, and is also closely related to inflammatory factors, tumor-related factors, etc. The Pi3k-akt signaling pathway is the upstream pathway of autophagy, and the hif-1 signaling pathway is not only the key pathway of hypoxia but also closely related to the damage of mitochondria by alcohol and its metabolites. This reveals that Huhuangan Liver Protecting Decoction is very likely to restore the intestinal barrier through the HIF-1 and pi3k-akt signaling pathways, improve mitochondrial function, affect autophagy, thereby preventing and treating acute alcoholic liver injury, and having a truncating effect on the development of alcoholic liver disease.

[0010] Professor Zhang Yongsheng believes that in the early stage of the disease, alcohol heat injures body fluids, alcohol dampness stagnates the spleen and stomach, and alcohol toxin disrupts the liver qi. Excessive drinking causes the pathogenic factors of alcohol toxin, dampness, and heat to soak the intestines and stomach, resulting in the failure of the middle earth to function properly, the disharmony of the middle jiao pivot, the disorder of ascending and descending, the stagnation of qi movement, the increasing of dampness stagnation, the accumulation of phlegm, the further obstruction of qi movement, the stagnation of blood circulation due to qi stagnation, the internal retention of stasis, and the combination of qi, phlegm, and blood, which knots under the hypochondrium and forms alcoholic mass. He proposed that the fundamental cause of alcoholic liver injury is excessive drinking, and the internal factor is congenital endowment deficiency and weakness of the spleen and stomach.

[0011] Specifically, the etiology and pathogenesis of alcoholic liver injury lie in:

[0012] (1) The liver fails to disperse and dredge, and the liver yin is deficient

[0013] The liver belongs to wood and is responsible for dispersing and dredging. When the liver disperses and dredges properly, the qi movement of the whole body is smooth, the five zang-organs are harmonious, the emotions are comfortable, the spleen and stomach function normally in transportation and transformation, and the body fluids are distributed orderly. The poisonous heat and dampness of alcohol accumulate in the liver and gallbladder, blocking the qi movement, leading to the stagnation of liver qi, and symptoms such as hypochondriac pain, abdominal distension, and bitter taste in the mouth will appear after a long time. At the same time, the disorder of the liver's dispersing and dredging function will also lead to the obstruction of blood circulation, forming qi stagnation and blood stasis, and liver mass and alcoholic mass will appear after a long time. Li Shizhen once said in "Yaolong Xiaopin": "Excessive drinking disrupts the mind, consumes blood, damages the stomach, generates damp phlegm, and promotes the desire for fire, causing various diseases." Alcohol is pungent, hot, dry, and fierce, and is easy to burn the liver yin, further damaging the liver.

[0014] (2)Weakness of the spleen and stomach

[0015] The spleen and stomach are the foundation of acquired constitution and the source of qi and blood production. The spleen and stomach provide qi, blood, and body fluids for the five zang-organs and six fu-organs by transporting and transforming refined substances of food and drink, enabling the normal functioning of the zang-fu organs. The normal operation of the liver's dispersing and discharging function depends on the support of the spleen and stomach. At the same time, alcohol is likely to damage the spleen and stomach, leading to the endogenous generation of dampness turbidity, further affecting the qi and blood circulation.

[0016] Currently, the incidence of alcoholic liver disease is increasing year by year. Drugs with good effects of protecting the liver and relieving alcohol intoxication, alleviating fatty degeneration, and having small toxic and side effects are still to be developed. Therefore, the inventor created the Huhu Huanggan Decoction, which has the functions of strengthening the spleen and nourishing yin, promoting blood circulation and qi movement, relieving alcohol intoxication and resolving fat. Its treatment principle is different from that of previous generations.

[0017] It can be seen from the pharmacological effects of each herb in the Huhu Huanggan Decoction. This formula has the effects of anti-inflammatory and antioxidant, improving the intestinal flora, and restoring the intestinal barrier function. This is also consistent with one of the pathogenesis mechanisms of alcoholic liver disease - the liver-intestine axis. Summary of the Invention

[0018] In view of the deficiencies of the prior art, the present invention provides a traditional Chinese medicine composition, preparation, and its application for treating alcoholic liver injury. The traditional Chinese medicine composition for treating alcoholic liver injury is the Huhu Huanggan Decoction, which adds nourishing yin and promoting blood circulation within the current mainstream treatment concept of "gentle diuresis for promoting dampness elimination". This formula has the effects of anti-inflammatory and antioxidant, improving the intestinal flora, and restoring the intestinal barrier function, and the whole formula has the functions of strengthening the spleen and nourishing yin, promoting blood circulation and qi movement, relieving alcohol intoxication and resolving fat.

[0019] In order to achieve the above object, the present invention adopts the following technical solutions:

[0020] A traditional Chinese medicine composition for treating alcoholic liver injury, the active ingredient raw materials of the traditional Chinese medicine composition include the following raw materials in parts by weight: 5 - 15 parts of Dendrobium officinale, 6 - 10 parts of Curcuma longa, 10 - 20 parts of Pueraria lobata, 6 - 12 parts of Six-Flavored Medicinal Leaven, and 2 - 6 parts of Amomum kravanh.

[0021] Preferably, the active ingredient raw materials of the traditional Chinese medicine composition include the following raw materials in parts by weight: 9 - 11 parts of Dendrobium officinale, 7 - 9 parts of Curcuma longa, 10 - 12 parts of Pueraria lobata, 9 - 12 parts of Six-Flavored Medicinal Leaven, and 5 - 6 parts of Amomum kravanh.

[0022] Preferably, the active ingredient raw materials of the traditional Chinese medicine composition include the following raw materials in parts by weight: 10 parts of Dendrobium officinale, 8 parts of Curcuma longa, 10 parts of Pueraria lobata, 12 parts of Six-Flavored Medicinal Leaven, and 6 parts of Amomum kravanh.

[0023] Preferably, the traditional Chinese medicine composition further includes adding other traditional Chinese medicine extracts and / or adding other raw materials to enhance or assist the functions of strengthening the spleen and nourishing yin, promoting blood circulation and qi movement, relieving alcohol intoxication and resolving fat of the above components.

[0024] Furthermore, the present invention also provides a traditional Chinese medicine composition preparation for treating alcoholic liver injury, which comprises the above-mentioned traditional Chinese medicine composition and / or adding medicinal excipients and / or adding food additives for improving the stability of the dosage form.

[0025] Preferably, the traditional Chinese medicine composition preparation is a tablet, chewable tablet, powder, powder for oral use, granule, instant granule, capsule, paste, pill or liquid preparation.

[0026] Furthermore, the present invention also provides the use of the above-mentioned traditional Chinese medicine composition or the traditional Chinese medicine composition preparation in the preparation of drugs for alcoholic liver injury.

[0027] Preferably, the above use is used to relieve the pathological damage of the patient's liver and the symptoms of liver fat deposition.

[0028] Preferably, the above use is used to protect the gastrointestinal mucosa of the patient.

[0029] Explanation of the formula of the present invention:

[0030] Dendrobium officinale was first recorded in "Shennong Ben Cao Jing" in the Eastern Han Dynasty: "Dendrobium officinale tastes sweet and flat, mainly treating internal injuries, dispelling rheumatism, lowering qi, tonifying the five internal organs, emaciation due to consumptive disease, strengthening yin, long-term use can thicken the stomach and intestines, lighten the body and prolong life." "Ming Yi Bie Lu" supplemented that Dendrobium officinale can also "regulate the stomach qi, grow muscles, expel skin problems, relieve heat rash qi, and relieve pain and cold weakness in the feet and knees". Miao Xiyong started from the "classic" and believed that "Dendrobium officinale receives the yang qi in the middle of the earth... its qi is thin and its taste is thick, it is yin in yang. It enters the Yangming meridian of the foot, the Shaoyin meridian of the foot, and also enters the Shaoyin meridian of the hand... It can remove the dampness of the spleen and stomach meridians." Zhang Jingyue believed that: Dendrobium officinale can be used to remove the fire of the spleen and stomach, relieve noisy hunger and heat accumulation in the nutrient system, relieve palpitations and calm the mind. Zhang Shanlei in "Ben Cao Zheng Yi" said: "Dendrobium officinale clears heat and reduces qi, specifically purges the deficiency fire of the lung and stomach, and its taste is not thin. Therefore, it is a product that benefits the stomach and strengthens yin." "Ben Cao Zheng Yi" first mentioned the relevant records of Dendrobium officinale: "It must be the one with dark green skin color, firm texture, sticky tongue when chewed raw, and thick and slightly sweet taste as the top grade, named Tiepi fresh Dendrobium." Zhang Jingyue in "Jing Yue Quan Shu · Consumption" in the Ming Dynasty said: "Wine is originally a crazy drug that greatly damages the true yin." The nature of wine is pungent and hot, which is easy to damage body fluids and damage the true yin. Dendrobium officinale nourishes yin and promotes fluid production. Generally speaking, Dendrobium officinale can remove the dampness and heat in the spleen and stomach and the upward reversal of qi caused by drinking, and can also tonify deficiency, strengthen yin, and thicken the stomach and intestines to prevent the injury of alcohol toxicity.

[0031] Curcuma longa was first recorded in *Newly Revised Materia Medica*. It is pungent, bitter, extremely cold, and non-toxic. It is mainly used to treat abdominal mass, zhù (chronic infectious disease), reverse qi, lower qi and break blood stasis, dispel wind-heat, and eliminate carbuncles and swelling. *Compendium of Materia Medica for Ready Reference* states that it enters the spleen and liver meridians and can regulate qi in blood. *Speculations on Materia Medica* mentions that it enters the spleen meridian, so it can treat abdominal distension, and enters the liver meridian, so it can promote blood circulation and disperse stasis. *Feng's Secret Recipes* believes that curcuma longa is yin within yang, descending in nature, and can quickly lower qi and break all stagnant qi. In the Qing Dynasty, Wu Tang believed that "the liver governs blood and the collaterals also govern blood", and put forward the idea of "treating the liver must treat the collaterals". Curcuma longa can just lower the reversed qi after drinking and dissolve the stasis formed over a long illness.

[0032] When used together, they can not only remove dampness and purge heat, promote qi movement and activate blood circulation, but also tonify deficiency and nourish yin without promoting dampness.

[0033] Pueraria lobata was first recorded in *Shennong's Classic of Materia Medica*. It is sweet and neutral. It is mainly used to treat thirst, high fever of the whole body, vomiting, various arthralgias, promote the production of yin qi, and detoxify various poisons. *Treatise on Medicinal Properties* states that pueraria lobata "detoxifies alcohol and relieves vexation and thirst"; *Supplements to Materia Medica* records that pueraria lobata can "detoxify alcohol" and treat "body heat, red complexion due to alcohol". *Treasury of Health Preservation* also states that pueraria lobata "treats thirst due to spleen deficiency, clears stomach heat, detoxifies alcohol, and promotes the circulation of the Foot-Yangming Meridian." Pueraria lobata relieves alcohol intoxication because its clear and rising property can disperse the alcohol toxin accumulated in the Yangming Meridian. Chen Shiduo in *Secret Records of the Stone Chamber* pointed out that although pueraria lobata has the effect of relieving alcohol intoxication, it is difficult to completely cure the diseases caused by excessive drinking: "The syndrome of hematochezia is mostly caused by excessive drinking. Using products to relieve alcohol intoxication may achieve success, but it is not always the case. However, kudzu root should not be taken in large amounts...." Its ascending and dispersing property can cause excessive depletion of body fluids, so it should not be taken for a long time. In this formula, pueraria lobata promotes qi movement, relieves muscle and purges heat to disperse alcohol toxin, eliminates vexation and stops vomiting to calm the stomach qi. It is used together with dendrobium officinale to prevent excessive damage to body fluids.

[0034] Medicated leaven was recorded in *Correct Materia Medica* in the Ming Dynasty: "It tastes sweet, has a neutral qi, and is stir-fried yellow before being used as medicine. It is good at assisting the middle-jiao earth viscera, strengthening the spleen and warming the stomach, promoting digestion and lowering qi, regulating qi and relieving stagnation, and expelling phlegm accumulation." Ye Tianshi recorded in *Explanation of Materia Medica* that it "has a warm qi, tastes pungent and sweet, and is non-toxic. It mainly treats stagnant food and water, accumulation and binding, strengthens the spleen and warms the stomach." On the one hand, medicated leaven has the effect of strengthening the spleen and warming the stomach. When used together with dendrobium officinale, it nourishes yin and protects the stomach to prevent alcohol from damaging the spleen and stomach; on the other hand, medicated leaven has a warm qi and disperses the liver. Together with pueraria lobata and amomum kravanh, it can assist the smooth flow of liver qi to relieve stagnation.

[0035] Amomum kravanh was recorded in *Explanation of Materia Medica*: "It has a very warm qi, tastes pungent, and is non-toxic. It mainly treats accumulated cold qi, stops vomiting and counterflow nausea, promotes digestion and lowers qi." Amomum kravanh is warm in nature and enters the Foot-Jueyin Liver Meridian and the Hand-Shaoyang Sanjiao Meridian, and has a good qi-promoting function.

[0036] In the formula, dendrobium officinale and medicated leaven strengthen the spleen, protect the stomach and nourish yin, curcuma longa breaks qi and promotes blood circulation, pueraria lobata and amomum kravanh promote qi movement and activate collaterals. They not only remove dampness and purge heat, promote qi movement and activate blood circulation, but also tonify deficiency and nourish yin without promoting dampness. The whole formula together exerts the effects of strengthening the spleen and nourishing yin, promoting blood circulation and qi movement, relieving alcohol intoxication and resolving fat.

[0037] Modern pharmacological analysis of the formula of this invention:

[0038] Dendrobium officinale has a variety of biological activities, such as anti-inflammatory, antioxidant, immunomodulatory and cardiovascular protective effects. Dendrobium officinale polysaccharide can bind to the surface receptors of immune cells, affect the functions of macrophages and lymphocytes, and thus play an immunomodulatory role. Some studies have shown that Dendrobium officinale can play antioxidant and anti-aging roles by reducing the levels of reactive oxygen species (ROS) and malondialdehyde (MDA), increasing the activities of catalase (CAT), superoxide dismutase (SOD) and the content of glutathione (GSH). At the same time, it has functions such as regulating the intestinal flora, promoting gastric acid secretion, improving gastrointestinal motility, and inhibiting intestinal mucosal damage. It has certain effects on different biological models of gastric mucosal damage.

[0039] Curcuma longa has various functions such as antibacterial, antioxidant, anti-apoptotic, anti-tumor and anti-metastatic effects. Its main component, curcumin, can play a variety of beneficial roles in the gastrointestinal tract, such as preventing reflux esophagitis, Barrett's esophagus and gastric mucosal damage caused by non-steroidal anti-inflammatory drugs (NSAIDs). The antioxidant effect of curcumin is mainly reflected in scavenging reactive oxygen free radicals, inhibiting lipid peroxidation reactions, increasing the activities of superoxide dismutase (SOD) and catalase (CAT), etc., and having a significant inhibitory effect on lecithin lipid peroxidation and induced DNA oxidative damage.

[0040] Pueraria lobata polysaccharide can reduce the concentrations of isovaleric acid and isobutyric acid in the cecum and enrich the diversity of the intestinal flora. At the same time, the active components of total flavonoids in Pueraria lobata, such as puerarin, daidzein, and daidzin, have good anti-inflammatory, antioxidant, antibacterial, and anti-tumor activities, and can promote the growth of immune cells and immune organs. A number of studies have shown that total flavonoids in Pueraria lobata can significantly improve alcoholic fatty liver and have the effect of relieving alcohol. Puerarin can protect against acute alcohol poisoning and acute liver injury by inhibiting the release of β-EP and free radicals and resisting the increase in blood viscosity caused by alcohol.

[0041] Six-Flavored Medicinal Leaven has the functions of improving intestinal function, regulating the intestinal flora, and maintaining the integrity of the intestinal structure. Some studies have shown that Six-Flavored Medicinal Leaven can inhibit the expression of adipocyte-specific mRNA to reduce fat production, restore lipid metabolism, and promote fatty acid utilization and other mechanisms to relieve non-alcoholic fatty liver and control type 2 diabetes, etc.

[0042] Amomum kravanh contains rich volatile oil components, has pharmacological activities such as kidney protection and anti-tumor, and can increase the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in renal tissues. The flavonoid content also has pharmacological effects such as antioxidant activity, anti-tuberculosis and liver protection. It can enhance the activity of alcohol dehydrogenase, thus achieving the prevention and treatment of alcoholic hepatitis.

[0043] The Huguang Hugan Decoction of the present invention can significantly reduce the levels of serum ALT, AST and TG in mice, reduce the content of MDA and the levels of IL-1β, IL-6, TNF-α in the liver, increase the level of SOD, reduce liver fat deposition in mice, relieve liver pathological damage in mice, reduce the levels of oxidative stress and inflammatory factors, and has a good protective effect on the liver with alcoholic liver injury. Description of the Drawings

[0044] Figure 1 It is the liver index of mice in the beneficial effect study. Among them, A is the single 56° alcohol gavage model group, and B is the NIAAA model group.

[0045] Figure 2 It is the serum ALT level of mice in the beneficial effect study. Among them, A is the single 56° alcohol gavage model group, and B is the NIAAA model group.

[0046] Figure 3 It is the serum AST level of mice in the beneficial effect study. Among them, A is the single 56° alcohol gavage model group, and B is the NIAAA model group.

[0047] Figure 4 It is the serum TG level of mice in the beneficial effect study. Among them, A is the single 56° alcohol gavage model group, and B is the NIAAA model group.

[0048] Figure 5 It is the serum SOD and MDA levels of mice in the beneficial effect study. Among them, A is the single 56° alcohol gavage model group, and B is the NIAAA model group.

[0049] Figure 6 It is the serum IL-1β, IL-6, TNF-α levels of mice in the beneficial effect study. Among them, A is the single 56° alcohol gavage model group, and B is the NIAAA model group.

[0050] Figure 7 It is the HE staining of liver tissues of mice under the single 56° alcohol gavage model in the beneficial effect study. Among them, a is the blank group 1, b is the model group 1, c is the positive control group 1 (glutathione group), and d is the treatment group 1 (Huguang Hugan Decoction group).

[0051] Figure 8 It is the HE staining of liver tissues of mice under the NIAAA model in the beneficial effect study. Among them, a is the blank group 2, b is the model group 2, c is the positive control group 2 (glutathione group), and d is the treatment group 2 (Huguang Hugan Decoction group).

[0052] Figure 9For the Oil Red O staining of mouse liver tissue in the 56° alcohol gavage model once in the beneficial effect study, where a is the blank group 1, b is the model group 1, c is the positive control group 1 (glutathione group), and d is the treatment group 1 (Huhuang Hugan Decoction group).

[0053] Figure 10 For the Oil Red O staining of mouse liver tissue in the NIAAA model in the beneficial effect study, where a is the blank group 2, b is the model group 2, c is the positive control group 2 (glutathione group), and d is the treatment group 2 (Huhuang Hugan Decoction group).

[0054] Figure 11 For the PAS staining of mouse liver tissue in the 56° alcohol gavage model once in the beneficial effect study, where a is the blank group 1, b is the model group 1, c is the positive control group 1 (glutathione group), and d is the treatment group 1 (Huhuang Hugan Decoction group).

[0055] Figure 12 For the PAS staining of mouse liver tissue in the NIAAA model in the beneficial effect study, where a is the blank group 2, b is the model group 2, c is the positive control group 2 (glutathione group), and d is the treatment group 2 (Huhuang Hugan Decoction group).

[0056] Figure 13 For the liver index of the orthogonal experiment.

[0057] Figure 14 For the serum TG level of mice in the orthogonal experiment.

[0058] Figure 15 For the serum ALT level of mice in the orthogonal experiment.

[0059] Figure 16 For the serum AST level of mice in the orthogonal experiment.

[0060] Figures 17 - 19 For the HE staining of mouse liver tissue in the orthogonal experiment.

[0061] Figure 20 For the serum SOD level of mice in the orthogonal experiment.

[0062] Figure 21 For the serum MDA level of mice in the orthogonal experiment. Specific embodiments

[0063] Combined with the embodiments of the present invention below, the technical solutions in the embodiments will be clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0064] Example 1

[0065] The formula of Huhuangan Liver Protecting Decoction is: 10g of Dendrobium officinale, 8g of Curcuma longa, 10g of Pueraria lobata, 12g of Six-Flavored Medicinal Leaven, and 6g of Amomum kravanh.

[0066] This example is the best compatibility, which is verified by orthogonal experiment as follows:

[0067] I. Test content

[0068] The prescription of Huhuangan Liver Protecting Decoction was optimized by orthogonal design to analyze the best dosage of each drug in this prescription in order to achieve the best therapeutic effect. Considering the recommended dosages of each drug in the pharmacopoeia, the five-flavored drugs in the formula were taken as the research objects, and the comprehensive scores were obtained with ALT, AST, SOD, and MDA as the evaluation indexes. The best drug combination of Huhuangan Liver Protecting Decoction was optimized according to the L18(3^5) orthogonal experiment design. The factor levels are shown in Table 1.

[0069] Table 1

[0070]

[0071] II. Research methods

[0072] 1. Experimental animals

[0073] 200 healthy SPF-grade male C57BL / 6 mice, 6 - 8 weeks old, with body weight (20 ± 5) g, were purchased from Shanghai SLAC Laboratory Animal Co., Ltd. (License number: SCXK(Shanghai)2017 - 0005). The mice were raised in the Animal Experiment Center of Zhejiang Chinese Medical University with good ventilation, temperature (23 ± 2)°C, relative humidity 40% - 60%, and relatively constant circadian rhythm (12h / 12h). Standard feed was provided, and the mice could eat and drink freely. After 1 week of adaptive feeding, the mice began to be modeled.

[0074] 2. Main reagents and instruments

[0075] 75% edible alcohol (Henan Hanshui Alcohol Co., Ltd.), each component drug of Huhuangan Liver Protecting Decoction, 10% neutral formaldehyde (provided by the school pathology laboratory), analytical pure sucrose (Xilong brand), malondialdehyde (MDA) biochemical kit (Nanjing Jiancheng), superoxide dismutase (SOD) biochemical kit (Nanjing Jiancheng), Shu Tai (prepared by the school animal house), high-speed centrifuge Centrifuge5424 (Eppendorf, Germany), tissue dehydrator Excelsior AS (Thermo Fisher Scientific, USA), embedding machine HistoCore Arcadia (Leica, Germany), paraffin slicer HM-355S (Thermo Fisher Scientific, USA), NX50 cryostat (Epredia, USA), staining machine (Thermo Fisher Scientific, USA), automatic biochemical analyzer (HITACHI, Japan), digital pathological slice scanning system VS120-S6-W (Olympus, Japan)

[0076] 3. Orthogonal array and grouping

[0077] A total of 20 groups including 18 groups of Huhuangan Liver Protecting Decoction with different dose combinations + blank group + model group, with 10 mice in each group. The mice in the blank group were gavaged with an equal dose of normal saline (10 ml / kg) for 10 days + once with an equal dose of normal saline, and the mice in the model group were gavaged with an equal dose of normal saline (10 ml / kg) for 10 days + once with 56° edible alcohol (14 ml / kg). The mice in the remaining groups were gavaged with the corresponding converted dose of the drug component (10 ml / kg) for 10 days and then once with 56° edible alcohol (14 ml / kg). The orthogonal design table is shown in Table 2.

[0078] Table 2 Orthogonal design table

[0079]

[0080]

[0081] 4. Result indicators

[0082] The scoring results were based on 30% ALT + 30% AST + 20% SOD + 20% MDA as the test indicators, and the other indicators such as HE scoring were for reference.

[0083] 4.1 General conditions and liver index

[0084] Observe the diet, behavior, state, hair and death of the rats.

[0085] Mouse liver index:

[0086] In the formula:

[0087] W——Liver index (%);

[0088] m——Mouse liver mass (g);

[0089] M——Mouse body weight (g).

[0090] 4.2 Serological detection

[0091] 12 hours after the last gavage (fasting but not water deprivation for 12 hours), the mice were anesthetized with Zoletil, blood was collected from the eye socket, centrifuged at 3500 rpm for 15 min at room temperature, the supernatant was taken and aliquoted into 1.5 ml centrifuge tubes, and the levels of ALT, AST, TBIL, ALP, and TG were measured in the blood biochemical laboratory of the Animal Center of Zhejiang Chinese Medical University.

[0092] 4.3 Pathological staining

[0093] Hematoxylin-eosin (HE) staining:

[0094] (1) The liver tissue was fixed with 10% neutral formaldehyde for 48 hours, then rinsed in running water for 1 to 2 hours, dehydrated, cleared, and infiltrated with paraffin before embedding and block making;

[0095] (2) The liver tissue was paraffin-embedded to make 4-μm thick sections;

[0096] (3) Deparaffinization and hydration: The sections were treated in xylene I and II for 15 minutes each, followed by 3 minutes each in a series of alcohols with concentrations of 100%-100%-95%-85%-75%, and rinsed with running water for 1 minute;

[0097] (4) Stained with hematoxylin for 5 minutes and then rinsed with running water;

[0098] (5) Differentiated with 1% hydrochloric acid alcohol for 10 seconds and immediately rinsed with running water for 5 minutes;

[0099] (6) Rinsed with tap water for several minutes to blue back;

[0100] (7) Stained with eosin for 3 - 5 minutes and then rinsed with running water;

[0101] (8) Dehydration and clearing: Through a series of alcohols with concentrations of 75%-85%-95%-100%-100% for 1 minute each, and xylene II and I for 1 minute each;

[0102] (9) Sealed with neutral gum;

[0103] (10) The pathological changes of liver tissues in each group were preliminarily observed using an optical microscope, and then scanned into digital pathological sections.

[0104] 4.4 Determination of oxidative stress-related indicators

[0105] The livers of mice were taken to measure the levels of SOD and MDA, and the specific method referred to the kit of Nanjing Jiancheng Company.

[0106] 5. Statistical methods

[0107] SPSS 26.0 was used to analyze the data and GraphPad Prism 6.01 software was used to plot the graphs. The data results were all expressed as "mean±SD". One-way ANOVA or non-parametric test was used to analyze and evaluate the differences between the two groups. P<0.05 indicated significant differences, and P<0.01 indicated extremely significant differences.

[0108] The highest values of ALT, AST, and MDA in each group were set to 100, and the lowest value of SOD was set to 100. The data was scored, and the K value and R value of each column of a single index were calculated according to the orthogonal table. K1, K2, and K3 were the average scores of the data results at the 1st level, 2nd level, and 3rd level respectively. The lower the score, the better the curative effect. The total K value was calculated as 0.3×ALT score + 0.3×AST score + 0.2×MDA score + 0.2×SOD score. R was the range = (K2 - K1) / 4, and in this experiment, R represented the effect difference between the two levels.

[0109] III. Research results

[0110] 1. General conditions of mice and liver index

[0111] The liver indexes of the mice in the groups treated with Huhuangan Liver Protecting Decoction on the 1st, 4th, 7th, 9th, 11th, 12th, 13th, 14th, and 17th days were significantly improved compared with those in the model group (P<0.05), while the liver indexes of the mice in the remaining groups were not significantly improved (P>0.05). The results are as Figure 13 shown.

[0112] 2. Serum ALT, AST, and TG levels of mice

[0113] The serum TG, ALT, and AST levels of each group of mice are shown in Figures 14 - 16 Table 3.

[0114] Table 3

[0115]

[0116] *p<0.05, **p<0.01.

[0117] Table 3 - continued

[0118]

[0119] *p<0.05, **p<0.01.

[0120] 3. Results of pathological HE staining

[0121] The HE staining results of the liver tissues of each group of mice are shown in Figures 17 - 19 . In the model group, a small amount of tissue cavities, inflammatory cell accumulation, and a small number of hepatocyte death and swelling were visible in the HE-stained mouse liver. The treatment groups showed improvement compared with the model group.

[0122] 4. SOD and MDA levels in mouse liver tissues

[0123] The SOD and MDA levels in the liver tissues of each group of mice are shown in Figures 20 - 21 and Table 4.

[0124] Table 4

[0125]

[0126] *p < 0.05, **p < 0.01.

[0127] Table 4 - continued

[0128]

[0129] 5. Orthogonal result scoring

[0130] Orthogonal result analysis was performed based on the scores and transformed scores of ALT, AST, SOD, and MDA in each treatment group. The optimal doses of each drug for single indicators and the optimal drug dose for comprehensive indicators are shown in Tables 5 and 6.

[0131] Table 5 Orthogonal design results - various indicators

[0132]

[0133]

[0134] Table 6 Orthogonal design results

[0135]

[0136]

[0137] From the orthogonal results:

[0138] In terms of ALT level, the R value of Amomum kravanh > the R value of Six-Flavored Powder Medicinal Leaven > the R value of Curcuma longa > the R value of Pueraria lobata > the R value of Dendrobium officinale, indicating that in terms of improving ALT level, the efficacy of Amomum kravanh > Six-Flavored Powder Medicinal Leaven > Curcuma longa > Pueraria lobata > Dendrobium officinale.

[0139] In terms of AST level, the R value of Dendrobium officinale > the R value of Curcuma longa > the R value of Amomum kravanh > the R value of Pueraria lobata > the R value of Six-Flavored Powder Medicinal Leaven, indicating that in terms of improving AST level, the efficacy of Dendrobium officinale > Curcuma longa > Amomum kravanh > Pueraria lobata > Six-Flavored Powder Medicinal Leaven.

[0140] In terms of SOD level, the R value of Dendrobium officinale > the R value of Amomum kravanh > the R value of Pueraria lobata > the R value of Curcuma longa > the R value of Six-Flavored Medicinal Leaven, indicating that in terms of improving SOD level, the efficacy order is Dendrobium officinale > Amomum kravanh > Pueraria lobata > Curcuma longa > Six-Flavored Medicinal Leaven.

[0141] In terms of MDA level, the R value of Curcuma longa > the R value of Dendrobium officinale > the R value of Amomum kravanh > the R value of Pueraria lobata > the R value of Six-Flavored Medicinal Leaven, indicating that in terms of improving MDA level, the efficacy order is Curcuma longa > Dendrobium officinale > Amomum kravanh > Pueraria lobata > Six-Flavored Medicinal Leaven.

[0142] In terms of the total score evaluation, the R value of Pueraria lobata > the R value of Curcuma longa > the R value of Amomum kravanh > the R value of Six-Flavored Medicinal Leaven > the R value of Dendrobium officinale, indicating that in terms of the overall evaluation, the efficacy order is Pueraria lobata > Curcuma longa > Amomum kravanh > Six-Flavored Medicinal Leaven > Dendrobium officinale.

[0143] The final optimal compatibility dosage is: 10 g of Dendrobium officinale, 8 g of Curcuma longa, 10 g of Pueraria lobata, 12 g of Six-Flavored Medicinal Leaven, and 6 g of Amomum kravanh.

[0144] The prescription of the optimal compatibility (i.e., Example 1) was used for the study of the beneficial effects of the present invention:

[0145] I. Experimental methods

[0146] 1. Pharmacodynamic evaluation of Huhu Huanggan Decoction in preventing and treating acute alcoholic liver injury

[0147] 1.1 Experimental animals

[0148] 50 healthy SPF-grade male C57BL / 6 mice, 6 - 8 weeks old, with a body weight of (20 ± 5) g, were purchased from Shanghai Slake Laboratory Animal Co., Ltd. (license number: SCXK (Shanghai) 2017 - 0005). The mice were housed in the Animal Experiment Center of Zhejiang Chinese Medical University, with good ventilation, a temperature of (23 ± 2) °C, a relative humidity of 40% - 60%, a relatively constant circadian rhythm (12h / 12h), provided with standard feed, and allowed to eat and drink freely. After 1 week of adaptive feeding, the mice began to be modeled.

[0149] 1.2 Main reagents and instruments

[0150] 75% edible alcohol (Henan Hanshui Alcohol Co., Ltd.), each component drug of Huhuangan Liver Protecting Decoction, reduced glutathione (Approved by the State Food and Drug Administration of China, H20050667, Chongqing Yaoyou Pharmaceutical Co., Ltd.), high-fat feed (Mouse One - Mouse Two), Oil Red O kit (provided by the Pathology Laboratory of Zhejiang Chinese Medical University), PAS staining kit (provided by the Pathology Laboratory of Zhejiang Chinese Medical University), 10% neutral formaldehyde (provided by the Pathology Laboratory of Zhejiang Chinese Medical University), analytical pure 95% ethanol (Xilong brand), analytical pure sucrose (Xilong brand), Shu Tai (prepared in the animal house of Zhejiang Chinese Medical University), mouse interleukin - 6 (IL - 6) ELISA scientific research kit (Aifang Biotech), mouse interleukin - 1β (IL - 1β) ELISA scientific research kit (Aifang Biotech), mouse tumor necrosis factor - α (TNF - α) ELISA scientific research kit (Aifang Biotech), malondialdehyde MDA biochemical kit (Nanjing Jiancheng), superoxide dismutase SOD biochemical kit (Nanjing Jiancheng), high - speed centrifuge Centrifuge5424 (Eppendorf, Germany), tissue dehydrator Excelsior AS (Thermo Fisher, USA), embedding machine HistoCore Arcadia (Leica, Germany), paraffin slicer HM - 355S (Thermo Fisher Scientific, USA), NX50 cryostat (Epredia, USA), staining machine (Thermo Fisher Scientific, USA), automatic biochemical analyzer (Hitachi, Japan), digital pathological slice scanning system VS120 - S6 - W (Olympus, Japan).

[0151] 1.3 Animal grouping

[0152] The model was established by intragastric administration of 56° edible alcohol (14 ml / kg) once and 10 - day alcohol liquid diet + intragastric administration of ethanol at 5 g / kg once. The drug intervention was carried out by the "human - mouse" equivalent dose method with conventional drug administration, and intragastric administration at 10 ml / kg was continuously performed for one week.

[0153] (1) Single - dose intragastric administration of 56° alcohol model

[0154] Blank Group 1: Intragastric administration of normal saline at 10 ml / kg QD; after 10 days, intragastric administration of normal saline at 14 ml / kg once;

[0155] Model Group 1: During the model establishment process, the same grasping was carried out, and intragastric administration of normal saline at 10 ml / kg QD; after 10 days, intragastric administration of 56° edible alcohol at 14 ml / kg once;

[0156] Positive control group 1 (glutathione group): Reduced glutathione was administered by gavage at a dose of 10 ml / kg once daily (QD), and after 10 days, 56° edible alcohol was administered by gavage at a dose of 14 ml / kg once;

[0157] Treatment group 1 (Huhuang Hugan Decoction group): Huhuang Hugan Decoction was administered by gavage at a dose of 10 ml / kg QD; after 10 days, 56° edible alcohol was administered by gavage at a dose of 14 ml / kg once.

[0158] (2) NIAAA model group

[0159] Blank group 2: Fed with isocaloric control liquid diet and administered normal saline by gavage QD; after 10 days, maltose was administered by gavage at a dose of 9 g / kg once;

[0160] Model group 2: Fed with 5% liquid alcohol diet and administered normal saline by gavage QD; after 10 days, ethanol was administered by gavage at a dose of 5 g / kg once;

[0161] Positive control group 2 (glutathione group): Fed with 5% liquid alcohol diet and administered reduced glutathione by gavage at a dose of 10 ml / kg QD; after 10 days, ethanol was administered by gavage at a dose of 5 g / kg once;

[0162] Treatment group 2 (Huhuang Hugan Decoction group): Fed with 5% liquid alcohol diet and administered Huhuang Hugan Decoction by gavage at a dose of 10 ml / kg QD; after 10 days, ethanol was administered by gavage at a dose of 5 g / kg once.

[0163] 1.4 Evaluation methods

[0164] 1.4.1 General conditions and liver index

[0165] The diet, behavior, status, hair, and death of the rats were observed.

[0166] Mouse liver index:

[0167] In the formula:

[0168] W - liver index (%);

[0169] m - mouse liver mass (g);

[0170] M - mouse body weight (g).

[0171] 1.4.2 Serological detection

[0172] Twelve hours after the last gavage (fasting but not water deprivation for 12 hours), the rats were anesthetized with Zoletil, and blood was collected from the eyeballs. The blood was allowed to stand at room temperature and centrifuged at 3500 rpm for 15 minutes. The supernatant was aliquoted into 1.5 ml centrifuge tubes and the levels of ALT, AST, and TG were measured in the blood biochemical laboratory of the Animal Center of Zhejiang Chinese Medical University.

[0173] 1.4.3 Pathological staining

[0174] After the last treatment, mice in each group were fasted but not watered for 12 hours. After intraperitoneal injection of 1% sodium pentobarbital for anesthesia, blood was collected from the eyeball for serological index detection. Liver and ileum tissues were separated, rinsed with saline, and fixed with 4% paraformaldehyde for histopathological examination.

[0175] The liver and ileum tissues fixed in 4% neutral formalin were taken and paraffin sections were prepared. The liver and ileum pathology were observed by HE staining, and the liver glycogen was detected by PAS staining. The liver tissue fixed in 4% neutral formalin was dehydrated in different concentrations of sugar water, and then frozen sections were made for Oil Red O staining.

[0176] 1.4.4 Determination of oxidative stress-related indicators

[0177] Mouse livers were taken to determine the levels of SOD and MDA. The specific method was referred to the kit of Nanjing Jiancheng Company.

[0178] 1.4.5 Determination of inflammation-related factors

[0179] Take mouse liver to measure the levels of IL-1β, IL-6, and TNF-α. After cutting the specimen, weigh it. Add a certain amount of PBS, pH 7.4. Quickly freeze it with liquid nitrogen for later use. After the specimen is thawed, it still maintains a temperature of 2-8°C. Add a certain amount of PBS (PH7.4) and homogenize the specimen thoroughly by hand or with a homogenizer. Centrifuge for about 20 minutes (2000-3000 rpm). Collect the supernatant carefully. After packaging, a portion is for testing, and the rest is frozen for later use. For specific methods, please refer to the instructions for the relevant Elisa kit of Aifang Bio.

[0180] 1.5 Statistical methods

[0181] SPSS26.0 was used to analyze the data and GraphPad Prism 6.01 was used to draw the graphs. The data results were expressed as "mean±SD". One-way ANOVA or nonparametric test was used to evaluate the differences between the two groups, and P < 0.05 indicated a significant difference.

[0182] 2. Results

[0183] 1. General

[0184] During the experiment, a total of 2 mice died. One was from Model Group 1 and the other was from Model Group 2. The rats in the normal control group had shiny hair, were lively, had normal food intake and bowel movements, and showed no drowsiness. In the NIAAA model, the mice in the model group all showed listlessness, messy and dull fur, slow movement, dull reaction, reduced food intake, slow weight gain or even weight loss. The Huhuangan Liver Protecting Decoction group showed fewer of the above manifestations than the model group. The glutathione group showed a slight improvement compared to the model group. Gross observation of the liver: The liver in the model group was significantly darker in color and larger in volume than that in the normal control group. In the model of a single intragastric administration of 56° alcohol, after alcohol intragastric administration, all the mice in the model group showed staggering when walking and accelerated heartbeat. After 10 minutes, all the mice stopped moving. When the mice were turned over to their backs after 30 minutes, half of the mice could not turn themselves back to their backs autonomously. Four mice in the Huhuangan Liver Protecting Decoction group were still moving slowly after 10 minutes, and when the mice were turned over to their backs after 30 minutes, all the mice could turn themselves back to their backs autonomously. The situation in the glutathione group was roughly the same as that in the model group.

[0185] 2. Liver index

[0186] The liver index is shown in Figure 1 . In the model of a single intragastric administration of 56° alcohol, the liver index of the mice in the Huhuangan Liver Protecting Decoction group was significantly improved compared to the model group (P < 0.05), while there was no obvious improvement in the glutathione group (P > 0.05); in the NIAAA model, the liver indices of the mice in the Huhuangan Liver Protecting Decoction group and the glutathione group were significantly improved compared to the model group (P < 0.05).

[0187] 3. Serum ALT, AST, and TG levels

[0188] The serum ALT, AST, and TG levels of the mice in each group are shown in Figures 2 - 4 . Compared with the blank group, the levels of ALT, AST, and TG in the model group were significantly increased (P < 0.05); compared with the model group, the levels of ALT, AST, and TG in the glutathione group and the Huhuajiejiu Decoction group were significantly decreased (P < 0.05); under the NIAAA model, there were significant differences in the ALT and AST levels between the glutathione group and the Huhuajiejiu Decoction group (P < 0.05).

[0189] 4. MDA and SOD levels in liver tissue

[0190] The MDA and SOD levels in the liver tissue of the mice in each group are shown in Figure 5The levels of MDA and SOD in the liver tissues of the mice in the model group were significantly different from those in the blank group (P < 0.05); the levels of MDA and SOD in the liver tissues of the mice in the glutathione group and the Huhuangan Decoction group were significantly improved compared with those in the model group (P < 0.05); and in the NIAAA model, the improvement in the Huhuangan Decoction group was better than that in the glutathione group. In the single 56° alcohol gavage model, the Huhuangan Decoction was superior to the glutathione group in improving the MDA level.

[0191] 5. Levels of IL-1β, IL-6, and TNF-α in liver tissues

[0192] The levels of IL-1β, IL-6, and TNF-α in the liver tissues of each group of mice are shown in Figure 6 . The levels of IL-1β, IL-6, and TNF-α in the liver tissues of the mice in the model group were significantly increased compared with those in the blank group (P < 0.05), while those in the Huhuangan Decoction group were significantly decreased compared with those in the model group (P < 0.05).

[0193] 6. HE staining of liver tissues

[0194] The HE staining of the liver tissues of each group of mice is shown in Figures 7 - 8 . In the single 56° alcohol gavage model, a small number of tissue cavities, inflammatory cell accumulation, and a small number of hepatocyte deaths and swelling were visible in the livers of the mice in the model group with HE staining. The glutathione group and the Huhuangan Decoction group showed improvement compared with the model group. In the NIAAA model, a large number of tissue cavities, inflammatory cell accumulation, hepatocyte deaths, and swelling were visible in the livers of the mice in the model group with HE staining. The glutathione group and the Huhuangan Decoction group showed improvement compared with the model group, with the tissue cavities decreasing and becoming smaller, and the hepatocyte deaths and swelling decreasing. Moreover, no inflammatory cell accumulation was seen in the Huhuangan Decoction group.

[0195] 7. Oil red O staining of liver tissues

[0196] The HE staining of the liver tissues of each group of mice is shown in Figures 9 - 10 . In the single 56° alcohol gavage model and the NIAAA model, a significant increase in lipid droplets was visible in the livers of the mice in the model group with oil red O staining. The glutathione group and the Huhuangan Decoction group showed improvement compared with the model group.

[0197] 8. PAS staining of liver tissues

[0198] The PAS staining of the liver tissues of each group of mice is shown in Figures 11 - 12 . In the single 56° alcohol gavage model and the NIAAA model, a significant decrease in glycogen synthesis was visible in the livers of the mice in the model group with PAS staining. The glutathione group and the Huhuangan Decoction group showed improvement compared with the model group, and the glycogen synthesis distribution in the livers of the mice in the Huhuangan Decoction group was more uniform.

[0199] III. Analysis of research results

[0200] 1. General situation analysis

[0201] During the experiment, all the dead mice were from the model group and there were no operational mistakes. It can be seen that both glutathione and Huhuangan Decoction have the effect of reducing death caused by excessive alcohol intake. In the NIAAA model, during the feeding of alcohol-containing diet, the mice in the Huhuangan Decoction group were significantly better than those in the model group and the glutathione group in terms of body weight, spirit, hair, activity, etc. After the last alcohol gavage, the general condition of the mice in the Huhuangan Decoction group was also significantly better than that of the model group and the glutathione group. In the model of single-dose intragastric administration of 56° alcohol, the situation of the mice in the Huhuangan Decoction group was also better than that of the glutathione group and the model group. It can be seen that Huhuangan Decoction may affect alcohol metabolism and protect the gastrointestinal tract, etc., which is related to the functions of the main components in Huhuangan Decoction to protect the integrity of the gastrointestinal mucosa and affect the activity of alcohol-metabolizing enzymes. It is suggested that Huhuajiejiu Decoction can well relieve the damage of alcohol to the body.

[0202] 2. Analysis of liver index

[0203] The metabolism of alcohol mainly occurs in the liver. When it exceeds the liver's metabolic capacity, it will cause liver cell damage and trigger a series of chemical reactions (oxidative stress and lipid peroxidation reactions), resulting in organ enlargement. Therefore, the liver index is often used as an indicator to measure liver enlargement. In the model of single-dose intragastric administration of 56° alcohol, Huhuajiejiu Decoction effectively improved the liver index of mice (P < 0.05), while although there was improvement in the glutathione group, there was no statistical difference. In the NIAAA model, there were statistical differences in improving the liver index of mice in both the Huhuangan Decoction group and the glutathione group (P < 0.05). The different results of the two models may be due to the different amounts of alcohol administered by single gavage. The alcohol intake of the mice in the model of single-dose intragastric administration of 56° alcohol was higher than that of the mice in the NIAAA model, and the NIAAA model mice ate 5% alcohol-containing diet for 10 days, and may have had a certain tolerance to alcohol, and the function of glutathione affected by a single high-dose alcohol intake. It also indicates that the protective effect of Huhuangan Decoction on the liver is still relatively obvious under different alcohol intakes.

[0204] 3. Analysis of serum indicators

[0205] In the model of single-dose intragastric administration of 56° alcohol, both Huhuangan Decoction and glutathione effectively improved the serum ALT and AST levels of mice (P < 0.05), and there was no statistical difference between the two. In the NIAAA model, both Huhuangan Decoction and glutathione effectively improved the serum ALT and AST levels of mice (P < 0.05), and the curative effect of Huhuangan Decoction was better than that of glutathione (P < 0.05). Both of them could improve the serum TG level of mice (P < 0.05), but there was no statistical difference in both models.

[0206] This indicates that Huhuangan Liver Protecting Decoction not only has good effects on liver injury caused by a single dose of alcohol, but also shows excellent curative effects in cases of multiple alcohol consumptions, acute + chronic alcoholic liver injury. This may be related to the fact that the components in Huhuangan Liver Protecting Decoction can not only effectively protect hepatocytes from direct damage by alcohol and its metabolites, but also have the function of protecting gastrointestinal mucosa, which can well prevent the translocation of LPS to the liver and cause secondary liver damage.

[0207] 4. Analysis of SOD and MDA levels in liver tissue

[0208] SOD is the main antioxidant enzyme defense system in organisms, which can effectively scavenge free radicals generated during the redox reactions of the body, repair cell damage, and play a crucial role in maintaining the free radical balance in the body. The level of its activity can reflect the antioxidant level of hepatocytes; MDA is a lipid peroxidation product and is one of the indicators used to evaluate the degree of liver damage caused by oxidative stress. In the single-dose 56° alcohol gavage model and the NIAAA model, both Huhuangan Liver Protecting Decoction and glutathione can effectively improve the SOD and MDA levels in the liver tissue of mice, and Huhuangan Liver Protecting Decoction is superior to glutathione in improving the SOD and MDA levels in the liver tissue of mice. This indicates that Huhuangan Liver Protecting Decoction can effectively improve liver damage in mice caused by alcohol by increasing the SOD level and reducing the accumulation of MDA in liver tissue. It is confirmed that Huhuangan Liver Protecting Decoction has a good antioxidant stress effect.

[0209] 4. Analysis of IL-1β, IL-6, and TNF-α levels in liver tissue

[0210] IL-1β, IL-6, and TNF-α are all important pro-inflammatory cytokines, which play key roles in the inflammatory response. These cytokines can stimulate the activation and proliferation of immune cells and inflammatory cells, promote the release of inflammatory proteins and inflammatory mediators, and thus exacerbate the inflammatory response. In the single-dose 56° alcohol gavage model and the NIAAA model, Huhuangan Liver Protecting Decoction can well improve the levels of IL-1β, IL-6, and TNF-α in the livers of mice. The release of these pro-inflammatory cytokines will exacerbate the oxidative stress state, leading to an increase in the production of free radicals. This may inhibit the activity of SOD, reduce its ability to scavenge free radicals, and at the same time increase the production of MDA, exacerbating the lipid peroxidation reaction. Huhuangan Liver Protecting Decoction has well improved this situation. This indicates that Huhuangan Liver Protecting Decoction has a good anti-inflammatory effect.

[0211] 5. Analysis of pathological staining of mice

[0212] Judging from the HE staining, both Huhuangan Liver Protecting Decoction and glutathione have improved the liver tissue damage in mice, including hepatocyte swelling, degeneration and necrosis, and cellular inflammatory infiltration. At the same time, it can also be seen that the liver damage in mice of the NIAAA model is more severe than that in the single-dose 56° alcohol gavage model.

[0213] From Oil Red O staining, both Huhuangan Liver-Protecting Decoction and glutathione improved the lipid deposition in the liver tissues of mice, and the improvement effect of Huhuangan Liver-Protecting Decoction was more obvious under the NIAAA model.

[0214] From PAS staining, both Huhuangan Liver-Protecting Decoction and glutathione improved the glycogen synthesis in the liver tissues of mice, and the improvement effect of Huhuangan Liver-Protecting Decoction was more obvious.

[0215] This confirmed that Huhuangan Liver-Protecting Decoction had good effects in both the acute alcoholic liver injury mouse model and the chronic + acute alcoholic liver injury mouse model.

[0216] The above is the description of the embodiments of the present invention. Through the above description of the disclosed embodiments, those skilled in the art can implement or use the present invention. Various modifications to these embodiments will be obvious to those skilled in the art. The general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but will conform to the widest scope consistent with the principles and novel points disclosed herein.

Claims

1. A Chinese medicine composition for treating alcoholic liver damage, characterized in that: The active ingredient raw materials of the Chinese medicine composition include the following raw materials by weight: 5-15 servings of Dendrobium candidum, 6-10 portions of turmeric, 10-20 portions of Pueraria root, 6 to 12 servings of Liu Shen Qu, 2 to 6 portions of white cardamom.

2. A Chinese medicine composition for treating alcoholic liver damage according to claim 1, characterized in that: The active ingredient raw materials of the Chinese medicine composition include the following raw materials by weight: 9-11 parts of Dendrobium candidum, 7-9 portions of turmeric, 10-12 portions of Pueraria root, 9-12 servings of Liu Shen Qu, 5-6 portions of white cardamom.

3. A Chinese medicine composition for treating alcoholic liver damage according to claim 1, characterized in that: The active ingredient raw materials of the traditional Chinese medicine composition include the following raw materials in parts by weight: 10 parts of Dendrobium officinale, 8 parts of Curcuma longa, 10 parts of Pueraria root, 12 parts of Liushenqu, and 6 parts of White Cardamom.

4. A Chinese medicine composition for treating alcoholic liver damage according to any one of claims 1 to 3, characterized in that: The Chinese medicine composition may also include other Chinese medicine extracts and / or other raw materials to enhance or assist the spleen-strengthening and yin-nourishing, blood-activating and qi-promoting, alcohol-resolving and lipid-removing effects of the above-mentioned components.

5. A Chinese medicine composition preparation for treating alcoholic liver damage, characterized in that: The Chinese medicine composition preparation comprises the Chinese medicine composition according to any one of claims 1 to 4 and / or added pharmaceutical excipients and / or added food additives, which are used to improve the stability of the dosage form.

6. A Chinese medicine composition preparation for treating alcoholic liver damage according to claim 5, characterized in that: The Chinese medicine composition preparation is in the form of tablets, chewable tablets, powders, granules, granules, capsules, ointments, pills or liquid preparations.

7. Use of the Chinese medicine composition according to any one of claims 1 to 4 or the Chinese medicine composition preparation according to any one of claims 5 to 6 in the preparation of a drug for alcoholic liver injury.

8. The use according to claim 7, characterized in that: The application is used to alleviate the patient's liver pathological damage and liver fat deposition symptoms.

9. The use according to claim 7, characterized in that: The application is used to protect the gastrointestinal mucosa of patients.

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