Charcoal medicine with effect of treating alcoholic liver injury

Abstract

The invention discloses a charcoal medicine with an effect of treating alcoholic liver injury, belongs to the technical field of medicines, relates to the charcoal medicine with the effect of treating the alcoholic liver injury, and in particular relates to a novel application of charcoal of achyranthes bidentata in preparation of a medicine for treating the alcoholic liver injury. Animal pharmacological experiments show that the charred achyranthes bidentata has a definite protection effect on alcoholic liver injury, and has the effects of relieving liver pathological injury, resisting inflammation and resisting oxidation.

Description

Technical Field

[0001] The invention relates to the medical field, and in particular to a new application of Achyranthes bidentata charcoal in the medical field. Background Art

[0002] Alcoholic liver injury is a series of liver damage caused by excessive accumulation of ethanol in the body due to acute or chronic long-term drinking, including hepatic fatty degeneration, fatty hepatitis, fibrosis, etc. If effective intervention measures are not taken, it will further develop into hepatocellular carcinoma. In severe cases, it can lead to death of patients, placing a great burden on social economy and medical care. At present, the drug treatment for severe alcoholic liver injury mainly relies on glucocorticoids, but its efficacy is limited. Alcohol abstinence is still the cornerstone of alcoholic liver disease treatment, but its recidivism rate is very high, and efficient targeted treatment for alcoholic liver injury is still under research and exploration. Due to the lack of effective drug treatment for alcoholic liver disease, there is an urgent need to develop new treatment methods and clinical drugs.

[0003] Achyranthes bidentata is the dried root of Achyranthes bidentata Bl., a plant of the Amaranthaceae family. It is bitter, sweet, sour and flat in nature. It enters the liver and kidney meridians. It has the effects of removing blood stasis and dredging menstruation, nourishing the liver and kidneys, strengthening tendons and bones, promoting urination and relieving stranguria, and guiding blood downward. However, according to ancient records, carbonized Achyranthes bidentata can treat tooth scurvy and pain, as well as postpartum lochia, scrofula, malignant sores, lip tubercle and other diseases.

[0004] Whether Achyranthes bidentata charcoal has a protective effect on alcoholic liver damage has not been reported in modern research. Summary of the invention

[0005] In view of this, the purpose of the present invention is to provide a safe and efficient Chinese herbal charcoal medicine for treating alcoholic liver damage, so that it can be used alone or in combination with Achyranthes bidentata charcoal as the main ingredient in clinical practice.

[0006] Specifically, the present invention is achieved by using Achyranthes bidentata charcoal in the preparation of a drug for treating alcoholic liver damage, and the preparation of Achyranthes bidentata charcoal comprises the following steps:

[0007] Put the raw medicinal slices of Achyranthes bidentata into a crucible, wrap and seal the crucible mouth with tin foil, cover the crucible with a lid, put it into a muffle furnace and set a certain temperature for firing.

[0008] The invention provides a new use of Achyranthes bidentata charcoal with effective therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 Effects of Achyranthes bidentata charcoal on the appearance of liver tissue in each group of mice. (A) Blank group; (B) Model group; (C) Positive drug group; (D) Achyranthes bidentata charcoal high-dose group; (E) Achyranthes bidentata charcoal medium-dose group; (F) Achyranthes bidentata charcoal low-dose group.

[0010] Figure 2 HE staining of liver tissue sections of mice treated with Achyranthes bidentata charcoal in each group (×200). (A) Blank group; (B) Model group; (C) Positive drug group; (D) Achyranthes bidentata charcoal high-dose group; (E) Achyranthes bidentata charcoal medium-dose group; (F) Achyranthes bidentata charcoal low-dose group.

[0011] Figure 3 .The effect of Achyranthes bidentata charcoal on AST and ALT, liver function indicators in mouse serum. (A) ALT; (B) AST.

[0012] Figure 4 .Effect of Achyranthes bidentata charcoal on TG content in mouse liver tissue.

[0013] Figure 5 Effects of Achyranthes bidentata charcoal on the levels of inflammatory factors in the serum of mice with alcoholic liver injury. (A) Interleukin 6 (IL-6); (B) Tumor necrosis factor α (TNF-α); (C) Interleukin IL-1β (IL-1β); (D) Interleukin 10 (IL-10).

[0014] Figure 6 Effects of Achyranthes bidentata charcoal on oxidative stress in liver tissue of mice with alcoholic liver injury. (A) Superoxide dismutase (SOD); (B) Glutathione (GSH); (C) Malondialdehyde (MDA).

[0015] Figure 7 Effects of Achyranthes bidentata charcoal on HO-1 protein expression in liver tissue of mice with alcoholic liver injury. (A) Blank group; (B) Model group; (C) Positive drug group; (D) Achyranthes bidentata charcoal high-dose group; (E) Achyranthes bidentata charcoal medium-dose group; (F) Achyranthes bidentata charcoal low-dose group.

[0016] Figure 8 Effects of Achyranthes bidentata charcoal on the expression of Nrf2 protein in liver tissue of mice with alcoholic liver injury. (A) Blank group; (B) Model group; (C) Positive drug group; (D) Achyranthes bidentata charcoal high-dose group; (E) Achyranthes bidentata charcoal medium-dose group; (F) Achyranthes bidentata charcoal low-dose group. DETAILED DESCRIPTION

[0017] The technical solution of the present invention will be clearly and completely described below in conjunction with specific embodiments, but the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. All other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0018] Embodiment 1: A method for preparing Achyranthes bidentata charcoal (calcination method), comprising the following steps:

[0019] Put the raw herbal medicine slices of Achyranthes bidentata into a crucible, cover the crucible mouth with aluminum foil, cover the lid tightly, and put it in a muffle furnace for carbonization (calcination temperature 400℃, time 1h). The obtained Achyranthes bidentata charcoal is black inside and outside and has a uniform texture.

[0020] Example 2: Study on the effect of Achyranthes bidentata charcoal in treating alcoholic liver damage

[0021] 1. Experimental Animals

[0022] Male C57BL / 6J mice were SPF grade, weighing 22.0±2.0g. During the animal breeding period, the temperature of the breeding room was maintained between 25℃-26℃, and the relative humidity was maintained at 55%-65%, with 12h light and dark alternation. During this period, food, water and bedding were changed in time to ensure the normal physiological environment of the experimental animals. All mice needed to be adaptively fed for 6 days before the formal experiment.

[0023] 2. Drug administration, modeling and sampling of experimental animals

[0024] Thirty-six male C57BL / 6J mice were randomly divided into blank control group, model group, positive drug bifendate group, Achyranthes bidentata charcoal high dose (5mg / kg), medium dose (2.5mg / kg), low dose (1.25mg / kg) administration group, a total of six groups, six mice in each group. After 5 days of adaptive feeding, each Achyranthes bidentata charcoal administration group was gavaged with Achyranthes bidentata charcoal solution of different concentrations, with a gavage volume of 0.1mL / 10g, and the model group and blank control group were gavaged with an equal volume of deionized water, and the drug was continuously administered for 6 days. Modeling began 2 hours after the last administration: except for the blank group gavaged with deionized water, the other groups were gavaged with 50% ethanol solution (12mL / kg), once every 12 hours, for 2 consecutive times, and the samples were collected 12 hours after the last modeling.

[0025] 12 hours after the last modeling, the eyeballs of mice in each group were removed for blood collection. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum and the content of triglyceride (TG) in liver tissue homogenate were detected according to the relevant instructions; the pathological damage of liver tissue was observed by H&E sections; ELISA was used to detect tumor necrosis factor α (TNF-α); interleukin 6 (IL-6); interleukin IL-1β (IL-1β); interleukin 10 (IL-10) in serum; superoxide dismutase (SOD); glutathione (GSH); malondialdehyde (MDA) in liver tissue were detected according to BCA protein quantification method; immunohistochemical staining was used to analyze the protein expression of Nrf2 and HO-1 in liver tissue.

[0026] 3. Evaluation of the therapeutic effect of Achyranthes bidentata charcoal on mice with alcoholic liver injury

[0027] During the experiment, the mice in the blank control group were very active overall, with normal drinking and eating, basically stable body weight, shiny and neat fur, normal bowel movements without diarrhea, etc.; after alcohol modeling, the mice in the model group showed obvious mental depression, significantly reduced activity, reduced food intake, increased secretions from the corners of the eyes, loss of luster in the skin and hair, and reduced body weight; after alcohol modeling, the mice in the Achyranthes bidentata charcoal-treated group and the positive control bifendate group also showed decreased fur brightness, activity, and food intake, but compared with the model group, their overall mental state was significantly improved.

[0028] Pathological manifestations of mouse liver Figure 1 As shown in the figure, the liver surface of mice in the blank group was smooth, bright red, with soft and slippery edges, and easy to peel off from surrounding organs and tissues. The liver of the alcohol model group was yellow in color, with severe adhesion to surrounding tissues, some granular protrusions on the surface, and some tissue edges were tough. The liver tissues of mice in the bifendate positive drug intervention group and the Achyranthes bidentata charcoal administration groups also had different degrees of pathological changes, but compared with the model group, they were all significantly improved.

[0029] The results of HE staining of liver tissues of mice in each group are shown in Figure 2 As shown in the figure, the liver lobule structure of the blank control group was intact, with clear boundaries, and the liver cords were arranged radially around. No inflammatory infiltration or fat vacuoles were observed. In the model group, fatty degeneration was more serious in most areas of the liver tissue, with a large number of fat vacuoles, disordered liver lobule structure, and hepatocyte necrosis and hemorrhagic lesions. The liver tissue damage of mice in the positive drug and Achyranthes bidentata charcoal intervention groups was significantly alleviated, the liver lobule structure was clear, the arrangement was relatively neat, and the fatty degeneration was significantly reduced. Moreover, among the different dose groups of Achyranthes bidentata charcoal, the liver tissue damage was significantly alleviated at low doses.

[0030] 4. Effect of Achyranthes bidentata charcoal on liver function indicators AST and ALT in mouse serum

[0031] like Figure 3 As shown in the figure, the ALT and AST levels in the serum of mice in the model group increased significantly compared with the blank group. Compared with the model group, the ALT and AST levels in the high, medium, low doses of Achyranthes bidentata charcoal and the positive drug group decreased significantly, and showed significant differences.

[0032] 5. Effect of Achyranthes bidentata charcoal on TG content in mouse liver tissue

[0033] The results of TG content detection in liver tissue homogenate of mice in each group are as follows Figure 4 : Compared with the blank control group, the TG content in the liver tissue of the model group increased significantly. The TG level in the liver tissue of the bifendate group and the high, medium and low dose groups of Achyranthes bidentata carbon decreased significantly.

[0034] 6. Effect of Achyranthes bidentata charcoal on the levels of inflammatory factors in mouse serum

[0035] Combined with previous related studies, interleukin 6 (IL-6); tumor necrosis factor α (TNF-α); interleukin 1β (IL-1β); and interleukin 10 (IL-10) were selected for detection. Figure 5 As shown in the results, after ALD was induced, the levels of proinflammatory factors IL-6, TNF-α, and IL-1β increased, and the level of anti-inflammatory factor IL-10 decreased. Compared with the model group, the group treated with Achyranthes bidentata charcoal significantly alleviated this phenomenon. This confirms that Achyranthes bidentata charcoal has good liver protection activity in the alcoholic liver injury model, and its pharmacological and pharmacodynamic mechanism may be related to the inhibition of inflammatory response.

[0036] 7. Effect of Achyranthes bidentata charcoal on oxidative stress in mouse liver tissue

[0037] like Figure 6 As shown in the results, after inducing ALD, the activities of SOD and GSH in liver tissue decreased significantly compared with the blank group; compared with the model group, the group treated with Achyranthes bidentata charcoal significantly increased the activity values ​​of SOD and GSH. At the same time, it promoted the removal of lipid peroxidation products, or inhibited the generation of MDA, reducing the level of oxidative stress.

[0038] 8. Effect of Achyranthes bidentata charcoal on the expression of HO-1 and Nrf2 proteins in liver tissue of mice in each group

[0039] like Figure 7 and Figure 8 The results of immunohistochemical staining for HO-1 and Nrf2 protein expression in liver tissue are shown in Figure 2. Compared with the blank control group, the expression of HO-1 and Nrf2 in liver tissue of mice in the model group was significantly reduced. Compared with the model group, the Achyranthes bidentata charcoal-treated groups and the positive drug group were able to significantly promote the expression of HO-1 and Nrf2 proteins in liver tissue of mice.

Claims

1. Use of Achyranthes bidentata charcoal in the preparation of drugs for the treatment of alcoholic liver damage.

2. The use according to claim 1, characterized in that The Achyranthes bidentata charcoal is used alone.

3. The use according to claim 1, characterized in that It is mainly used in combination with Achyranthes bidentata charcoal.

4. The use according to claim 1, characterized in that The Achyranthes bidentata charcoal and pharmaceutically acceptable auxiliary materials are prepared into a clinical preparation.

5. The use according to claim 4, characterized in that The clinical preparations include, but are not limited to, injections, oral preparations, external preparations, and various dosage forms for intracavitary administration.

6. A method for preparing the Achyranthes bidentata charcoal according to any one of claims 1 to 5, characterized in that: The Achyranthes bidentata is prepared by charcoal calcination.

7. The preparation method according to claim 6, characterized in that: The preparation method comprises the following steps: taking the raw medicinal pieces of Achyranthes bidentata, sealing the pieces, and firing the pieces at a certain temperature by a calcination method.

8. The preparation method according to claim 6, characterized in that: The preparation method comprises the following steps: sealing the raw medicinal slices of Achyranthes bidentata and placing them in a muffle furnace for carbonization (calcination temperature is 400° C. and time is 1 hour).

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