Viral or alcoholic liver disease relieving natural product based on ruta graveolens extract

Abstract

The invention provides a novel formula for relieving virus or alcoholic hepatitis and application of the formula, the formula is composed of the ruta graveolens extract or harmine, the ruta graveolens extract or harmine is successfully applied, a brand-new way is provided for coping with increasingly serious virus or alcoholic hepatitis problems, and the formula is a novel formula for relieving virus or alcoholic hepatitis. The natural product provided by the invention shows good liver protection activity in the research of relieving viruses or alcoholic hepatitis in vivo and in vitro.

Description

Technical Field

[0001] The invention relates to the use of a Rue extract and a composition based thereon in preparing a medicine for alleviating viral and alcoholic liver diseases, belonging to the field of medicines. Background Art

[0002] The liver is the largest organ in the human body. Its functions include: immune response, physiological metabolism, bile production, synthesis of anticoagulants, maintaining constant blood sugar, storing fat-soluble vitamins, phagocytizing damaged red blood cells and white blood cells, and participating in drug metabolism. If the liver is damaged and causes inflammation and necrosis of liver cells, the disease is called hepatitis. Hepatitis can generally be divided into: viral hepatitis, alcoholic hepatitis, chemical hepatitis and fatty hepatitis. The current cause of hepatitis among Chinese citizens is mainly viral, among which hepatitis B virus (HBV) infection is the most common cause. However, in the past few decades, the pattern of liver disease in my country has been gradually changing. With the establishment and promotion of the hepatitis B vaccine immunization expansion program, the number of people infected with hepatitis B virus has dropped significantly, while the number of people suffering from alcoholic cirrhosis is increasing.

[0003] Viral hepatitis refers to inflammation of the liver caused by the hepatitis virus invading the body and multiplying in liver cells. Symptoms include fatigue, loss of appetite, nausea, vomiting, etc. In severe cases, more critical symptoms such as fever or jaundice may occur. In rare cases, fulminant hepatitis with a high risk of death may occur. Viral hepatitis refers to a disease caused by inflammation of the liver after viral infection. It can currently be divided into five types: A, B, C, D, and E. The more common types are A, B, and C. Hepatitis A and E are transmitted through fecal-oral transmission, and most of the infection will not develop into chronic hepatitis. Hepatitis B, C, and D are contact infections transmitted through blood and body fluids. The special thing about hepatitis D is that it needs to coexist with hepatitis B to have the ability to infect and reproduce. Clinically, hepatitis D can be divided into two types. The first is simultaneous infection, which means that the patient is infected with acute hepatitis B virus and hepatitis D virus at the same time; the second is additional infection, which means that the patient is first infected with hepatitis B virus and then infected with hepatitis D virus. Usually, this type of infection has a poor prognosis and may develop into chronic hepatitis D, which also shows the importance of hepatitis B virus. In the treatment of viral hepatitis, there is currently no treatment for hepatitis A and E, and you can only rest more and wait for them to heal on their own. Hepatitis B and hepatitis D are mostly treated with interferon, which cannot be cured but can relieve inflammatory reactions. Drugs used to fight hepatitis B virus include entecavir, lamivudine, adefovir, telbivudine, etc. However, when taking drugs, the hepatitis B virus gene mutates, resulting in viral mutants, making the drugs unable to effectively inhibit the replication of the virus, that is, the originally effective drugs cannot be effective due to the occurrence of drug resistance. There are currently oral direct-acting antiviral (DAA) drugs available for hepatitis C, and about 90% of patients are completely cured after using them.

[0004] Although hepatitis B virus infection is the most common cause of liver cirrhosis in my country, the consumption of alcoholic beverages in my country has increased significantly in recent years, which has also increased the incidence of alcoholic liver disease. In 2014, the World Health Organization (WHO) published a report that Europe has the highest alcohol consumption and the highest incidence of alcoholic liver disease (ALD). However, from 1990 to 2014, alcohol consumption in Europe has been declining. However, in the past 25 years, Asia, especially China and India, has increased significantly. The World Health Organization's global drinking database shows that China's annual per capita alcohol consumption has increased more than 20 times in the past 50 years, and the proportion of excessive drinking has increased nearly 70 times.

[0005] Alcoholic liver disease includes a variety of liver diseases, such as alcoholic hepatitis, alcoholic liver steatosis, alcoholic steatohepatitis and alcoholic cirrhosis. Although drinking is a common behavior of human beings, different ethnic regions have different drinking cultures. However, various complications of alcoholic liver disease can also occur due to excessive drinking. In many countries, alcoholism has become a major problem. Studies have pointed out that chronic, heavy drinking is defined as consuming more than 40 grams of pure alcohol per day (equivalent to 375 ml of 13% alcohol by volume wine, or more than 1 liter of 5% alcohol by volume beer, converted to 50° liquor, that is, a daily consumption of more than 80 ml) for a continuous period of time (1 year), which greatly increases the risk of alcoholic liver disease.

[0006] After long-term alcohol intake, the biochemical and pathological damage caused in the human body is very complex, and the most affected tissue is the main tissue of alcohol metabolism - the liver. Liver damage caused by long-term alcohol intake includes: fatty liver, alcoholic hepatitis and cirrhosis, etc. If not treated in time, it will cause liver cancer. Studies have shown that patients with alcoholic hepatitis are susceptible to viral invasion and complications of viral hepatitis, among which alcoholic hepatitis has the highest incidence of hepatitis B and C; although chronic viral hepatitis mostly does not have obvious symptoms, it may also be stimulated and activated under certain circumstances such as overwork and alcoholism, causing the liver's inflammation index to continue to rise, and even evolve into cirrhosis and liver cancer. After patients with viral chronic hepatitis consume alcohol, liver disease will also deteriorate rapidly, increasing the probability of complications and death. Therefore, in areas of my country where the incidence of viral chronic hepatitis is high, more attention should be paid to liver disease problems caused by alcohol intake. In terms of drug and health product development, new products need to be developed for both viral hepatitis and alcoholic liver disease.

[0007] Ruta graveolens L. is a perennial herb belonging to the Sapindales order. Its seeds contain β-carboline alkaloids such as harmine, among which a specific inhibitor of human matrix metalloproteinase-3 (MMP-3) has been identified. The present invention further verifies the ability of Ruta graveolens extracts containing harmine and related alkaloids and other active ingredients to inhibit hepatitis B virus and hepatitis C virus and provide liver protection in an animal model of alcohol-induced liver injury. Although β-carboline alkaloids have good antimicrobial growth ability, there are also reports that excessive concentrations can also cause cytotoxicity to eukaryotic organisms. Therefore, the present invention uses Ruta graveolens extracts to test their anti-hepatitis B and C virus and liver protection activities.

[0008] The present invention first collects clinical serum from patients with positive hepatitis B and C as the virus source. In order to test the broad efficacy and universality of Rue extract against hepatitis B and C, HepaRG cell line is used as a virus replication tool. HepaRG is a liver cancer cell line isolated from a female patient with chronic hepatitis C virus infection, and is one of the few available cell line models susceptible to hepatitis virus infection. HepaRG cells have one or two nuclei, and their chromosome mutations are limited: including an extra chromosome 7 and a translocation phenomenon affecting chromosomes 12 and 22 (including the loss of the 12p region). Unlike the HepG2 liver cancer cell line used in the past, the amount of cytochrome P450 and liver nuclear receptors expressed by HepaRG cells is similar to that of primary normal human liver cells in culture, and can be relatively stable for more than six weeks. If DMSO is removed from the culture medium, the expression of P450 will be reduced, but the expression of liver-specific transcription factors and transporters will not be affected. The method for evaluating protection against alcoholic liver damage is to use the Lieber-DeCarli liquid alcohol feed to induce alcoholic fatty liver damage in C57BL / 6 (B6) mice in an experimental model, with the goal of establishing a formula that can prevent infectious hepatitis viruses and alcoholic liver disease that can cause significant harm to the human population. Summary of the invention

[0009] The purpose of the present invention is to provide a new use of natural rue extract, specifically, the use of the extract in preparing medicines for treating hepatitis B and C and alcoholic liver disease.

[0010] The use of the rue extract provided by the present invention is to prepare a drug for alleviating viral or alcoholic hepatitis. The rue extract is obtained by crushing the rue plant, extracting it with acid, and then precipitating the alkaloids with alkali.

[0011] The alkaloids are washed with alkali to obtain the Rutaecarpa refined extract.

[0012] Wherein, the acid is acetic acid, and the alkali is sodium bicarbonate (baking soda).

[0013] Wherein, the viral hepatitis refers to hepatitis B or hepatitis C.

[0014] The rue extract is derived from alkaloids and other effective substances extracted from mature seeds of Ruta plants of the Rutaceae family.

[0015] The Rue extract is prepared by the following method: crushing the seeds into coarse powder, adding 3 times the weight of 30% acetic acid solution, ultrasonically oscillating for 15 minutes, and then standing at room temperature for 48 hours, extruding and filtering the dough-like mixture, and then ultrasonically oscillating the residue with twice the weight of 30% acetic acid solution for 15 minutes, and repeating the filtration step once. Slowly add a large amount of sodium carbonate (pH 8) to the clarified liquid while stirring until the solution becomes turbid, stand at 4°C for 24 hours, suck off the supernatant and collect the precipitate (mainly containing alkaloids). The collected light yellow crystals are soluble in 60°C water and 100% acetic acid (1:3, v / v) (defined as "Rue crude extract" in the present invention); after the Rue crude extract is dissolved in dilute acetic acid, it is slowly titrated with sodium bicarbonate to pH 7.1 to 7.6 (usually pH 7.5), and the "refined extract" of the Rue extract can be further precipitated. The collected refined extract usually contains 95% harmine and 3% dihydroharmine.

[0016] The alkaloid-rich "crude extract" or "refined extract" of Rue can be washed with 3% ammonia water to prepare high-purity free alkaloids. The free alkaloids can be further processed into hydrochloride precipitates soluble in water by existing methods, separated, dried and weighed separately. The hydrochloride precipitates of alkaloids can be dissolved in water or directly formulated in various culture media at different concentrations (μg / ml).

[0017] In addition, the present invention also provides a use of a product for alleviating viral or alcoholic hepatitis, which is used for preparing a drug for alleviating viral or alcoholic hepatitis, and the product contains harmaline.

[0018] Wherein, the Rutaecarpa extract or the product contains 0.1 to 20 micrograms of harmine per milliliter. Preferably, the Rutaecarpa extract or the product contains 1 to 10 micrograms of harmine per milliliter.

[0019] The product comprises harmaline and another alkaline biological substance soluble in acetic acid aqueous solution, the alkaline biological substance being an alkaloid, for example, dictamnine, γ-fagarine, 4-methoxyfurano[2,3-B]quinoline-8-ol (robustine), zanthosimuline, toddaquinoline, etc.; coumarin, for example, toddalolactone, etc.; and purine, for example, adenine, guanine, hypoxanthine, xanthine, theophylline, etc.

[0020] One of the main effective alkaloids in Rutaecarpa identified by the present invention is harmine. It is previously known that high concentrations of harmine are toxic to mice and humans. According to reports, the intravenous lethal dose 50% (LD50) is 38 mg per kilogram of body weight, and humans will experience adverse symptoms when they ingest 3 mg per kilogram of body weight of harmine. Since the effective value of the present invention is at the μM level, and the literature estimates that the intestinal absorption rate of harmine in humans is 94.2%, it is very logical that oral administration can reach an effective dose without causing side effects.

[0021] The present invention also provides a natural ingredient composition for alleviating hepatitis B and C and alcoholic liver disease, wherein the composition contains rue extract as an active ingredient and is prepared by adding pharmaceutically acceptable auxiliary materials, adjuvants or auxiliary ingredients.

[0022] The pharmaceutical composition of the present invention is a preparation prepared from the following raw materials in the following weight ratio:

[0023] 1 part of crude rue extract or 0.01-0.1 part of refined rue extract.

[0024] Wherein, the medicament is an external preparation, an oral preparation or an injection preparation.

[0025] The raw material of the present invention, rue, is recorded in the book "Tacuinum sanitatis" adapted from the Arabic "health maintenance" manual in the 11th century in the Middle Ages in Europe. It has the function of improving eyesight and removing flatulence. Catholic priests even use rue branches for sprinkling holy water to enhance the function of removing evil spirits. It is a folk medicinal health plant that has been used for thousands of years.

[0026] In 2013, my country's National Health and Family Planning Commission stated that there are 350 million hepatitis B virus carriers in the world, of which China accounts for about 93 million people, and there are as many as 100,000 new infections each year, which has an "extremely serious" impact on China. In addition to continuously strengthening hospital infection prevention and control in medical and health institutions, comprehensively carrying out blood hepatitis virus nucleic acid testing in blood stations, and successfully developing domestic long-acting interferon and the world's first hepatitis E vaccine, the current situation of viral hepatitis prevention and control in my country is still severe, and the long-term accumulation of chronic viral hepatitis patients is large. Some antiviral treatment drugs are expensive, the drug accessibility is poor, and there is also a problem of drug resistance after long-term use. Therefore, the development of related drugs needs to be strengthened. At present, there are reports on the use of natural products for hepatitis B virus infectious diseases, such as patent number: CN102885856B, invention name: Production method for producing hepatitis B treatment drugs using American cockroach extracts. The application discloses a production method for producing hepatitis B treatment drugs using American cockroach extracts, which can improve the cure rate of hepatitis B. However, there is a lack of prior art records of natural products similar to the present invention that can target both type B and hepatitis B viruses.

[0027] Although everyone knows that excessive drinking can damage the liver, according to statistics from the World Health Organization in 2016, the number of deaths caused by excessive alcohol intake is as high as three million, which is equivalent to one in every twenty deaths caused by excessive drinking. However, unlike cirrhosis caused by viral hepatitis, there is still a lack of therapeutic drugs that are effective and can be recommended for alcoholic liver disease. "Guidelines for the Prevention and Treatment of Alcoholic Liver Disease (2018 Update)". Therefore, there is basically no wishful thinking of "getting an injection and taking medicine when you are sick". Therefore, the search for natural products for the treatment of alcoholic liver disease is also common in the literature. For example, some studies have pointed out that the ethanol extract of Ganoderma lucidum fruiting bodies rich in triterpenoid components has the effects of enhancing antioxidant enzymes and avoiding cell apoptosis. Animal experiments have confirmed that it can effectively reduce liver damage caused by alcohol. The Rue extract of the present invention not only has antiviral activity, but also has a strong protective effect on alcoholic liver disease. There is no relevant literature report on such natural products that target both viruses and alcoholic liver disease.

[0028] Obviously, for those skilled in the art, based on the above contents of the present invention, according to the common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modifications, replacements or changes can be made.

[0029] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the protection scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The cell morphology of HepaRG cells observed under an optical microscope is shown in Figure A. The cell morphology of HepaRG cells when undifferentiated is relatively simple when observed under an optical microscope; and in Figure B, approximately 50% of HepaRG cells differentiate into bile duct cell morphology and liver cell morphology under an optical microscope, with the ratio of the two types of cells being approximately 1:1.

[0031] Figure 2 The results of hematoxylin andeosin stain (H&E stain) of HepaRG cell samples before and after virus infection are shown. A shows the morphology of HepaRG cells before differentiation. B shows the morphology of uninfected HepaRG cells after differentiation. The cells clustered in the central red circle are bile duct cell-like; the larger cells in the right red circle are liver cell-like. C shows the morphology of HepaRG cells infected with the virus after differentiation. The infected cells are two to three times larger than the surrounding cells. The arrows point out the liver cell-like cells in the process of ballooning degeneration.

[0032] Figure 3 Western blot analysis was used to confirm the inhibitory effect of the harmine component in Rutaecarpa extract on the expression of hepatitis B virus protein. DETAILED DESCRIPTION

[0033] The following specific pharmacodynamic tests prove that the present invention has a helpful effect on alleviating or inhibiting viral and alcoholic liver diseases.

[0034] Example 1 HepaRG cell culture and differentiation:

[0035] HepaRG cells are cultured in a special way to differentiate into liver cells and bile duct cells. First, 37°C containing 10% FBS, 5×10 -7 HepaRG cells were cultured in 75T culture flasks with 14 ml of Williams' E medium containing 1 M hydrocortisone and 5 μg / ml insulin, and placed in an incubator at 37°C and 5% CO2. The medium was changed every three to four days. When the cells filled the culture flask, the expansion culture could be performed. The cell supernatant was removed as much as possible with a micropipette, PBS was added to rinse the cells, and 3 ml of trypsin that had been warmed to 37°C was added and allowed to act for 3 minutes to make the cells float. The cell fluid was then aspirated and centrifuged. After centrifugation, the cells were seeded into four culture flasks to complete the expansion step.

[0036] For HepaRG cell differentiation, the concentration of hydrocortisone in the culture medium was adjusted to 5×10 -5 M, and add 2% DMSO to induce cell differentiation. When the undifferentiated HepaRG cells fill the culture flask (about two weeks), use a micropipette to remove as much supernatant as possible, rinse the cells with PBS, add 37°C HepaRG differentiation medium, replace the medium every three to four days and observe the cell differentiation. When the homogeneous cells show about 50% hepatocytes and bile duct-like morphology ( Figure 1 ) can be used for virus infection vaccination.

[0037] Example 2 Virus infection of HepaRG cells:

[0038] After HepaRG cells differentiated more than 30% for two weeks, 10% FBS in the culture medium was replaced with serum from hepatitis carriers. The supernatant was removed as much as possible with a micropipette, and the cells were rinsed with PBS. 37°C HepaRG virus infection medium (containing 20% ​​serum from virus carriers) was added. After 3-4 days, the culture medium was replaced back to HepaRG differentiation medium. Each time the culture medium was replaced, the culture medium was kept in a -20°C refrigerator.

[0039] The culture medium was prepared using plasma from carriers of hepatitis B or C virus. After culturing with differentiated HepaRG cells for three to four days, the differentiation medium was replaced. Each time the culture medium was changed, hepatitis B or C virus was tested by quantitative polymerase chain reaction (PCR) or quantitative reverse transcription PCR (RT-qPCR). The results showed that HepaRG cells could be successfully infected with the virus using this infection method. Not only did the HepaRG cells actively synthesize the hepatitis virus and release it into the culture medium, but compared with the uninfected group, it was found that since the hepatitis virus specifically infects liver cells, the infected group had significantly fewer liver cell-type cells, while bile duct cell-type cells had space to proliferate in large numbers. The large-scale proliferation of bile duct cell-type cells resulted in a higher proportion of this cell type, and the liver cell-like cells underwent a ballooning degeneration process. The HepaRG cells were two to three times larger than the surrounding cells and were almost bursting ( Figure 2 ), consistent with viral hepatitis.

[0040] Example 3 Effect of Rutaecarpa extract on HepaRG cells infected with hepatitis B virus:

[0041] The product of Qiagen (Hilden, Germany) was used. HBV RG PCR kit and its associated instrument quantify the amount of HBV released by HepaRG cells under different states of virus infection. After culturing differentiated HepaRG cells with culture medium prepared with plasma from HBV carriers for three to four days, the culture medium was replaced with differentiation medium prepared with fetal bovine serum and free of virus. Various drugs were added to the culture medium. After 72 hours, the culture supernatant was collected and centrifuged at 1,000 x g for 20 minutes. The supernatant was used to extract viral DNA using the QIAamp DSP Virus kit produced by Qiagen. The operation method of HBV RGPCR kit was to quantify the amount of hepatitis B virus in each treatment group three times and record the results. As shown in Table 1, both the crude extract of Rutaecarpa and the refined extract of Rutaecarpa could neutralize the virus, showing that they could significantly inhibit the synthesis of hepatitis B virus DNA, and their high concentration (10 μg / ml) was more effective than 3330 pg / ml of α-type interferon (IFN-2α) purchased from Sigma-Aldrich.

[0042] Table 1 Inhibitory effect of the extract obtained by the above disclosed method for preparing Rutaecarpa extract on hepatitis B virus

[0043]

[0044] Example 4 Effect of Rutaecarpa extract on HepaRG cells infected with hepatitis C virus:

[0045] Use Qiagen HCV RG RT-PCR kit and its associated instrument quantify the amount of HCV released by HepaRG cells under different states of virus infection. After three to four days of culture medium prepared with plasma from hepatitis C carriers and differentiated HepaRG cells, the culture medium was replaced with virus-free differentiation medium prepared with fetal bovine serum, to which various drugs were added. After 72 hours, the culture supernatant was collected and centrifuged at 1,000 x g for 20 minutes. The supernatant was used to extract viral RNA using the QIAamp DSP Virus kit produced by Qiagen. The operation mode of HCV RG RT-PCR kit was to quantify the amount of hepatitis C virus in each treatment group three times and record the results. As shown in Table 2, both the crude extract of Rutaecarpa and the refined extract of Rutaecarpa could neutralize the virus, and the high concentration (10 μg / ml) was more effective than 3330 pg / ml of α-type interferon (IFN-2α) purchased from Sigma-Aldrich.

[0046] Table 2: Inhibitory effect of extracts obtained according to the above disclosed method for preparing Rutaecarpa extract on hepatitis C virus

[0047]

[0048] Example 5 The present invention confirms the effect of the harmaline component in the Rutaecarpa extract on hepatitis viruses:

[0049] The protein in each group of HepaRG cells infected with HBV was extracted by preparing a denaturing protein sample buffer (DPSB) containing 3% sodium dodecylsulfate (SDS) in Tris-HCl (pH 6.8). After adding 80 μl of DPSB, the cells in the dish were quickly scraped back and forth with a sterile cell scraper until the cells were completely scraped off (the liquid was thick), and then the freeze-thaw process was performed at -80℃ and 99℃ to complete the protein extraction. After the protein was electrophoresed, the electrophoresis gel was first rinsed with phosphate buffered saline (PBS) containing 0.05% Tween20 for 15 minutes, and then transferred for 120 minutes. After the transfer was completed, it was rinsed with phosphate buffered saline (PBST) for 10 minutes three times, and then the first antibody against the surface protein of HBV was added and allowed to act at 4℃ for 16 hours. After washing with PBST for 10 minutes three times, the secondary antibody was added. After one hour at room temperature, the cells were washed with phosphate buffered saline containing 0.05% polyoxyethylene sorbitan monolaurate (Tween 20) for 10 minutes three times. Immobilon ECL Ultra Western HRP Substrate produced by Merck KGaA was added and the results were observed with a cold light imaging system and analyzed with ImageJ software. The size of S protein is about 24kDa, and the glycosylated S protein is 27kDa; the size of M protein is about 33kDa, and the glycosylated M protein is 36kDa; the size of L protein is about 39kDa, and the glycosylated L protein is 42kDa.

[0050] Western blot was used to detect the inhibitory effects of Rutaecarpa extract and Harmine on the protein expression of HBV in HepaRG cells. Rutaecarpa crude extract could inhibit the expression of viral protein by 16%, while 10μg / ml and 1μg / ml Harmine could inhibit the expression of HBV protein by about 50% respectively ( Figure 3 ). It can be inferred that harmaline in Rutaecarpa extract is the main hepatitis B virus inhibitory component.

[0051] Example 6: Effect of Rutaecarpa extract and harmaline on the alleviation of alcoholic liver disease:

[0052] Chronic liver damage by alcohol was studied using C57BL / 6 mice weighing about 20 grams. Alcoholic liver disease was induced by long-term feeding of Lieber–DeCarli alcohol-containing liquid diet. The mice were divided into five groups (10 mice in each group), including control group (C), alcohol group (E), alcohol and silymarin group (ES), alcohol and crude extract of Rutaecarpa group (ER), and alcohol and harmaline group (EH). Group C was given a normal liquid diet, while groups E, ES, ER, and EH were fed with Lieber–DeCarli liquid diet. Group ES was gavaged with 200 mg silymarin (Sigma-Aldrich) per kg of body weight per day, while groups ER and EH were gavaged with 1 mg crude extract of Rutaecarpa or 0.5 mg 99.8% harmaline per kg of body weight per day. The experimental period was 8 weeks, and then the activity of enzymes related to liver damage (aspartate transaminase, AST; glutamate aminotransferase, alanine transaminase, ALT) and myeloperoxidase (MPO) and components (triglyceride, TG; cholesterol, CH; glucose, BG) were measured.

[0053] The results of measuring the enzyme activity related to liver damage are shown in Tables 3 and 4, respectively. The results show that the crude extract of Rutaecarpa has the highest protective effect on alcoholic liver disease, especially the inflammatory indicator MPO (Table 3) and the blood sugar reduction phenomenon of alcohol reducing glucose regeneration genes and glycogen consumption (Table 4), which can almost return to normal levels.

[0054] Table 3: Effects of various treatments on serum glutamate aminotransferase (ALT), aspartate aminotransferase (AST) and MPO enzyme activity in liver of C57BL / 6 mice with alcohol-induced liver injury

[0055]

[0056] Table 4: Effects of various treatments on changes in triglyceride (TG), cholesterol (CH) and plasma glucose (BG) content in liver (mg per gram of liver) of C57BL / 6 mice with alcohol-induced liver damage

[0057]

[0058] The above embodiments fully demonstrate that the method of preparing Rutaecarpa extract and Echinops quinata extract provided by the present invention has the advantages of high efficiency, rapidity, low cost, and many optional combinations in the future compared with traditional new drug development, and has shown good alleviating activity in in vitro and in vivo studies on viral and alcoholic hepatitis.

Claims

1. A use of a Rutaecarpa extract for preparing a medicament for alleviating viral or alcoholic hepatitis, characterized in that: The rue extract is prepared by crushing the rue plant, extracting it with acid, and then precipitating the alkaloids with alkali.

2. The use according to claim 1, wherein the alkaloid is further titrated with alkali to obtain a refined extract of Rutaecarpa at a pH between 7.1 and 7.

6.

3. The use according to claim 2, wherein the Rutaecarpa refined extract contains 95% of harmine and 3% of dihydroharmine.

4. The method according to claim 1 or 2, wherein the acid is acetic acid and the base is sodium bicarbonate.

5. The use according to claim 1, wherein the viral hepatitis is hepatitis B or hepatitis C.

6. The use according to claim 1, wherein the Rutaecarpa extract contains 0.1 to 20 micrograms of harmine per milliliter.

7. The use according to claim 6, wherein the Rutaecarpa extract contains 1 to 10 micrograms of harmine per milliliter.

8. Use of a product for alleviating viral or alcoholic hepatitis, for preparing a drug for alleviating viral or alcoholic hepatitis, characterized in that: The preparation comprises harmine.

9. The use according to claim 8, wherein the preparation contains 0.1 to 20 micrograms of harmine per milliliter.

10. The use according to claim 8, wherein the preparation contains 1 to 10 micrograms of harmine per milliliter.

11. The use according to claim 8, wherein the viral hepatitis is hepatitis B or hepatitis C.

12. The use according to claim 8, in the relief of alcoholic hepatitis, the preparation further comprises another alkaline biological substance soluble in aqueous acetic acid solution.

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