Use of a chenopodium zollingeri extract in the preparation of a medicament for the treatment and / or prevention of non-alcoholic fatty liver disease

Abstract

This invention discloses the application of Artemisia argyi extract in the preparation of drugs for the treatment and / or prevention of non-alcoholic fatty liver disease, belonging to the field of traditional Chinese medicine extraction technology. The preparation method of the Artemisia argyi extract of this invention includes the following steps: mixing Artemisia argyi powder with a 60% (v / v) ethanol aqueous solution for extraction, separating the solid and liquid phases to obtain an ethanol extract; concentrating the ethanol extract to obtain a paste, drying it, and completing the preparation. The extraction process designed in this invention is simple and efficient, obtaining an extract rich in flavonoids, terpenes, phenols, and sesquiterpene lactones and other active ingredients from Artemisia argyi. In vitro and in vivo studies show that this extract can alleviate liver function and lipid abnormalities, reduce hepatic lipid deposition, and has relatively few toxic side effects. This invention not only provides a scientific basis for the rational development and utilization of characteristic medicinal resources, but also lays a theoretical foundation for traditional drugs to enter the market.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine extraction technology, and in particular to the application of an extract of Artemisia argyi in the preparation of drugs for the treatment and / or prevention of non-alcoholic fatty liver disease. Background Technology

[0002] Artemisia argyi is a semi-shrubby herbaceous plant belonging to the Artemisia genus of the Asteraceae family. It grows to a height of about 40 cm and is mostly found in mountain basins (desert areas) and terrace gullies, with an altitude range of 3200~3800 m.

[0003] Nonalcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome characterized by excessive lipid accumulation in hepatocytes, after excluding alcohol and other known liver-damaging factors (such as drugs, viral infections, and autoimmune etiologies). It may or may not be accompanied by simple steatosis and intralobular inflammation. The NAFLD spectrum ranges from simple fatty liver (SFL) to advanced nonalcoholic steatohepatitis (NASH), which can lead to liver fibrosis (LF), cirrhosis (LC), hepatocellular carcinoma, and liver failure. The development of NAFLD is driven by multiple mechanisms, including genetic factors, insulin resistance, lipotoxicity, mitochondrial dysfunction, endoplasmic reticulum stress, inflammation, gut microbiota dysbiosis, and adipose tissue dysfunction. Currently, the socioeconomic burden of NAFLD and its complications is increasingly heavy, and there are no approved specific treatments globally. Therefore, the development of innovative drugs for this disease has significant scientific and clinical value.

[0004] In recent years, the prevalence of NAFLD has continued to rise, posing a serious threat to human health. Traditional Chinese medicine compound formulas, with their multi-component and multi-target characteristics, have demonstrated unique advantages in the prevention and treatment of chronic metabolic diseases, representing a natural treasure trove for developing anti-NAFLD drugs. Summary of the Invention

[0005] The purpose of this invention is to provide an application of Artemisia argyi extract in the preparation of drugs for the treatment and / or prevention of non-alcoholic fatty liver disease, thereby addressing the aforementioned problems in the background art. The extraction process designed in this invention is simple and efficient, yielding an extract from Artemisia argyi rich in various active ingredients, including flavonoids, terpenes, phenols, and sesquiterpene lactones. In vitro and in vivo studies show that this extract can alleviate liver dysfunction and dyslipidemia, reduce hepatic lipid deposition, and has minimal toxic side effects. This invention not only provides a scientific basis for the rational development and utilization of unique medicinal resources but also lays a theoretical foundation for the market entry of traditional medicines.

[0006] To achieve the above objectives, the present invention provides the following technical solution: One of the technical solutions of the present invention is to provide the application of Artemisia argyi extract in the preparation of a drug for treating and / or preventing non-alcoholic fatty liver disease.

[0007] Preferably, the preparation method of the Artemisia argyi extract includes the following steps: The powdered Artemisia argyi was mixed with a 60% (v / v) ethanol aqueous solution for extraction, followed by solid-liquid separation to obtain the ethanol extract. The alcohol extract was concentrated to obtain an extract, which was then dried to obtain the Artemisia argyi extract.

[0008] Preferably, the ratio of the Artemisia argyi powder to the ethanol aqueous solution is 1g:15~25mL.

[0009] Preferably, the extraction time is 60-84 hours; The concentration temperature is 40~60℃ and the pressure is 0.07~1MPa.

[0010] Preferably, the drying is freeze drying; the freeze drying temperature is -43.4℃, the vacuum degree is 4.6Pa, and the time is 6~10h.

[0011] Preferably, the drug for treating and / or preventing non-alcoholic fatty liver disease is a drug that alleviates liver function and blood lipid abnormalities.

[0012] Preferably, the drug for treating and / or preventing non-alcoholic fatty liver disease is a drug that reduces lipid deposition in the liver.

[0013] The beneficial technical effects of the present invention are as follows: The extraction process for Artemisia annua extract designed in this invention is simple and efficient. Using a 60% (v / v) ethanol aqueous solution as the extraction solvent, an extract rich in flavonoids, terpenes, phenols, and sesquiterpene lactones was obtained from Artemisia annua. This invention is the first to propose the application of Artemisia annua extract in the preparation of drugs for the prevention and / or treatment of metabolic-related lipid disorders. In vitro and in vivo studies show that this extract can alleviate liver dysfunction and dyslipidemia, reduce hepatic lipid deposition, and has relatively few toxic side effects. This invention not only provides a scientific basis for the rational development and utilization of specialty medicinal resources but also lays a theoretical foundation for the market entry of traditional medicines.

[0014] In vitro experiments showed that the Artemisia annua extract could significantly reduce triglyceride (TG) and total cholesterol (TC) levels and effectively inhibit lipid accumulation. In vivo experiments further confirmed that the product could significantly reduce body weight, epididymal adipose tissue (EAT) weight, and anterior abdominal adipose tissue (AAT) weight in NAFLD model mice, and improve serum aspartate aminotransferase (AST), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglycerides (TG), total cholesterol (TC), and liver triglyceride (TG) and total cholesterol (TC) levels. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 The results show the effects of Artemisia argyi extract on HepG2 cell viability in Examples 1 and Comparative Examples 1-3; where A is Artemisia argyi aqueous extract, B is Artemisia argyi 30% alcohol extract, C is Artemisia argyi 60% alcohol extract, and D is Artemisia argyi 90% alcohol extract. Figure 2 The results show the effects of Artemisia argyi extract from Examples 1 and Comparative Examples 1-3 on TG expression in NAFLD cell models; where A is Comparative Example 3, B is Comparative Example 1, C is Example 1, and D is Comparative Example 2. Figure 3 The effect of 60% ethanol extract of Artemisia annua from Example 1 on cellular lipid accumulation is shown in Figure 1. A represents the Oil Red O staining results for the normal group, model group, low-dose group, medium-dose group, and high-dose group, respectively; B is a statistical graph of the Oil Red O staining results; and C represents the measurement results of intracellular TC and its content. Figure 4 The results of the experiment in Example 1 on the relief of HFD-induced NAFLD by 60% ethanol extract of Artemisia argyi; where AB are the images showing the changes in mouse body weight and liver ultrasound after drug administration; C is the statistical graph of mouse body weight change after drug administration; D is the statistical graph of cumulative food intake and body weight of mice after drug administration; E is the statistical graph of mouse body weight before dissection. Figure 5 The results of the experiment in Example 1 on the relief of HFD-induced NAFLD by 60% ethanol extract of Artemisia argyi; where A is a graph showing the changes in liver and epididymal fat weight in mice after administration; B is a statistical graph showing the changes in liver in mice after administration; C is the epididymal fat index in mice; D is the abdominal fat index in mice; EG is a graph showing the blood glucose test results. Figure 6The results of the 60% ethanol extract of Artemisia annua in Example 1 on the levels of lipid-related indicators in the serum and liver of mice induced by HFD are shown below. A represents the change in serum AST levels; B represents the change in serum ALT levels; C represents the change in serum LDL levels; D represents the change in serum HDL levels; E represents the change in serum TG levels; F represents the change in serum TC levels; G represents the change in liver TG levels; and H represents the change in liver TC levels. Figure 7 This document summarizes the pathological staining results for the normal group, model group, low-dose group, medium-dose group, high-dose group, and positive control group. A represents the staining results; B represents the statistics of Oil Red O staining results; C represents the statistics of HE staining results; and D represents the HE staining results of epididymal fat. Detailed Implementation

[0017] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention. It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the present invention.

[0018] Furthermore, regarding the numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0019] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. It should be noted that any aspects of this invention not described in detail are conventional practices in the art and are not the focus of this invention.

[0020] The terms “comprising,” “including,” “having,” “containing,” etc., used in this invention are all open-ended terms, meaning that they include but are not limited to.

[0021] This invention discloses a method for preparing Artemisia annua extract for preparing anti-NAFLD drugs, comprising the following steps: The powdered Artemisia argyi was mixed with a 60% (v / v) ethanol aqueous solution for extraction, followed by solid-liquid separation to obtain the ethanol extract. The alcohol extract was concentrated to obtain an extract, which was then dried to obtain the Artemisia argyi extract.

[0022] Furthermore, the ratio of the Artemisia argyi powder to the ethanol aqueous solution is 1g:15~25mL.

[0023] Furthermore, the ratio of the Artemisia argyi powder to the ethanol aqueous solution is 1g:20mL.

[0024] Furthermore, the extraction temperature is room temperature, and the time is 60-84 hours, preferably 72 hours.

[0025] Furthermore, the extraction process is carried out in a sealed environment to ensure that the ethanol does not evaporate.

[0026] Furthermore, the solid-liquid separation method is filtration.

[0027] Furthermore, the concentration is carried out using a rotary drum, preferably stopping when liquid begins to adhere to the drum wall.

[0028] Furthermore, the concentration temperature is 40~60℃, preferably 50℃, and the pressure is 0.07~1MPa, preferably 0.8MPa.

[0029] Furthermore, the drying process is freeze drying.

[0030] Furthermore, the freeze-drying temperature is -43.4℃, the vacuum degree is 4.6Pa, and the time is 6~10h, preferably 8h.

[0031] The present invention also discloses an Artemisia argyi extract prepared according to the above preparation method for the preparation of anti-NAFLD drugs.

[0032] The present invention also discloses the use of the above-mentioned Artemisia argyi extract in the preparation of a medicament for the treatment and / or prevention of metabolic-related fatty diseases.

[0033] Furthermore, the metabolic-related fatty diseases include liver disease or obesity.

[0034] Furthermore, the liver disease mentioned is non-alcoholic fatty liver disease (NAFLD).

[0035] In this invention, through in vitro screening, the optimal concentration of FFA (free fatty acids) for in vitro modeling was determined to be 0.5 mM, and the modeling time was 24 h. The Artemisia argyi extract (60% ethanol extract of Artemisia argyi) of this invention was found to be the extract with the strongest in vitro anti-NAFLD effect. The 60% ethanol extract of Artemisia argyi of this invention exerts its pharmacological effect in the in vitro NAFLD model by significantly reducing triglyceride and total cholesterol levels and inhibiting lipid accumulation. The 60% ethanol extract of Artemisia argyi can improve abnormal serum liver function and lipid-related indicators in mouse NAFLD models, exerting its anti-NAFLD pharmacological effect by reducing lipid deposition.

[0036] The preparation method of Artemisia argyi powder used in the following embodiments and comparative examples of the present invention is as follows: Take the whole plant of Artemisia argyi, dry it, cut it into pieces, and crush it into coarse powder.

[0037] Unless otherwise specified, "room temperature" in this invention refers to 10-30°C.

[0038] Unless otherwise specified, all materials and equipment used in this invention are commercially available products in the field.

[0039] To further illustrate the present invention, the following description, in conjunction with the accompanying drawings and embodiments, further explains the Artemisia argyi extract, its preparation method, and its application, but this should not be construed as limiting the scope of protection of the present invention.

[0040] Example 1 (60% alcohol extract) A method for preparing Artemisia annua extract, comprising the following steps: Accurately weigh 100g of Artemisia argyi powder and add it to a 60% ethanol aqueous solution at a ratio of 1g:20mL. After soaking at room temperature for 72h, filter to obtain the ethanol extract. Concentrate under reduced pressure using a rotary evaporator (50℃, 0.8MPa) to obtain an extract. Then freeze-dry (temperature -43.4℃, vacuum degree 4.6Pa, drying for 8h) to make powder, thus obtaining 60% ethanol extract of Artemisia argyi (yield 13.2%).

[0041] Comparative Example 1 (30% alcohol extract) A method for preparing Artemisia annua extract, comprising the following steps: Accurately weigh 100g of Artemisia annua powder and add it to a 30% ethanol aqueous solution at a ratio of 1g:20mL. After soaking at room temperature for 72h, filter to obtain the ethanol extract. Concentrate under reduced pressure using a rotary evaporator (50℃, 0.8MPa) to obtain the extract. Then, vacuum dry (dry at 45℃ for 4h) to make powder, thus obtaining 30% ethanol extract of Artemisia annua (yield of 15.97%).

[0042] Comparative Example 2 (90% alcohol extract) A method for preparing Artemisia annua extract, comprising the following steps: Accurately weigh 100g of Artemisia annua powder and add it to a 90% ethanol aqueous solution at a ratio of 1g:20mL. After soaking at room temperature for 72h, filter to obtain the ethanol extract. Concentrate under reduced pressure using a rotary evaporator (50℃, 0.8MPa) to obtain the extract. Then, vacuum dry (dry at 45℃ for 4h) to make powder, thus obtaining 90% ethanol extract of Artemisia annua (yield of 11.8%).

[0043] Comparative Example 3 (Water Extract) A method for preparing Artemisia annua extract, comprising the following steps: Accurately weigh 100g of Artemisia annua powder, add water at a ratio of 1g:20mL, soak at room temperature for 72h, filter to obtain water extract, concentrate under reduced pressure using a rotary oscillator (50℃, 0.8MPa) to obtain extract, and then vacuum dry (dry at 45℃ for 4h) to make powder, thus obtaining Artemisia annua water extract (yield 23.51%).

[0044] Effect verification 1. Cytotoxicity detection HepG2 cells in the logarithmic growth phase were used to prepare a density of 8×10⁻⁶ cells / year. 3 pcs·mL -1 Cell suspensions were seeded into 96-well plates. When cells reached a density of 70-80%, the old culture medium was discarded, and drug-containing culture medium containing Artemisia argyi extract prepared in Example 1 and Comparative Examples 1-3 (final concentrations of 500, 250, 125, 62.5, 31.75, 15.5, and 7.75 μg·mL, respectively) was added. A solvent control group (containing only solvent) and a blank control group (containing no cells) were also set up, with 6 replicates in each group. After culturing for 24 h, the culture medium was discarded, and the cells were washed twice with PBS. 100 μL of 10% CCK-8 reagent was added to each well, and the cells were incubated for 1 h. The absorbance (A) of each well was measured at 450 nm using a microplate reader, and the average value was used to calculate cell viability. The experiment was repeated three times in parallel. The toxicity of the drug to normal cells was then assessed. Results are as follows: Figure 1 As shown.

[0045] Figure 1 The results show the effects of Artemisia argyi extract on HepG2 cell viability in Examples 1 and Comparative Examples 1-3; where A is Artemisia argyi aqueous extract, B is Artemisia argyi 30% ethanol extract, C is Artemisia argyi 60% ethanol extract, and D is Artemisia argyi 90% ethanol extract.

[0046] in accordance with Figure 1 The experimental results determined the safe concentration ranges for each extract: 0–500 μg / mL for the aqueous extract of Artemisia argyi, 0–500 μg / mL for the 30% ethanol extract of Artemisia argyi, 0–1000 μg / mL for the 60% ethanol extract of Artemisia argyi, and 0–500 μg / mL for the 90% ethanol extract of Artemisia argyi. Subsequent experiments determined the appropriate dosage concentration based on these results.

[0047] 2. Determination of intracellular TG (triglycerides) and its content Cells were seeded and cultured in 6-well plates. Experimental groups included a control group, a model group (FFA, free fatty acids), and a treatment group (FFA + Artemisia argyi extract). After culturing according to the pre-selected modeling conditions (FFA concentration 0.5 mM, treatment for 24 h) and the drug concentration, the cell supernatant was discarded, cells were washed three times with PBS, and cells were collected by trypsin digestion. After centrifugation and discarding the supernatant, 150 μL of lysis buffer was added to lyse the cells. The cell lysate was collected and incubated on ice for 30 min. Intracellular triglyceride (TG) levels were measured according to the instructions of the corresponding kits. Significant changes in TG levels were observed at the cellular level. Results are as follows: Figure 2 As shown.

[0048] Figure 2 The results show the effect of Artemisia annua extract from Examples 1 and Comparative Examples 1-3 on TG expression in the NAFLD cell model; where A is Comparative Example 3, B is Comparative Example 1, C is Example 1, and D is Comparative Example 2 (Note: FFA is a modeling agent, free fatty acid, at a concentration of 0.5 mM). Different batches of samples were tested in groups A, B, C, and D, with a blank control group and a model group set up in each group.

[0049] Depend on Figure 2 It can be seen that after administering the Artemisia annua extracts of Examples 1 and Comparative Examples 1-3, the 60% ethanol extract of Artemisia annua from Example 1 significantly inhibited TG expression. The TG content in the model group reached 2.14 mmol / gprot, significantly increased compared to the blank group. However, after intervention with the 60% ethanol extract of Artemisia annua, the TG content in the treated groups decreased in a dose-dependent manner, and the TG content in the high-dose group was as low as 1.35 mmol / gprot, a highly statistically significant difference compared to the model group (P<0.01). In contrast, the aqueous extract, 30% ethanol extract, and 90% ethanol extract of Artemisia annua from Comparative Examples 1-3 did not significantly inhibit TG expression.

[0050] 3. Effects of 60% alcohol extract of Artemisia annua on lipid accumulation in vitro According to the grouping settings, place a coverslip at the bottom of the 6-well plate, take HepG2 cells in the logarithmic growth phase, digest them to prepare a single-cell suspension, count the cells and adjust the cell density to 4 × 10⁶ cells per well. 5One cell (containing 0.5 mL of culture medium) was seeded onto a coverslip. After the cells stabilized, 1.5 mL of culture medium was added to each well, and the cells were transferred to an incubator for further culture. After 24 hours of stable culture, the supernatant was aspirated. Incomplete culture medium was added to the control wells (normal group), the model group was given 0.5 mM FFA, and the drug-treated groups were diluted with 0.5 mM FFA solution. 60% ethanol extract of Artemisia argyi was administered at concentrations of 125 μg / mL (high-dose group), 31.75 μg / mL (medium-dose group), and 7.75 μg / mL (low-dose group), respectively. After treatment, cell morphology was observed, and Oil Red O staining was performed. The results are as follows: Figure 3 As shown.

[0051] Figure 3 The effect of 60% ethanol extract of Artemisia annua from Example 1 on cellular lipid accumulation is shown in Figure A, which shows the Oil Red O staining results of the normal group (CON), model group (FFA), low-dose group, medium-dose group, and high-dose group, respectively; Figure B is a statistical graph of the Oil Red O staining results.

[0052] 4. Determination of intracellular TC (total cholesterol) and its content. Cells were seeded and cultured in 6-well plates. Experimental groups included a control group, a model group (FFA, free fatty acids), and a treatment group (FFA + Artemisia argyi extract). After culturing according to the pre-selected modeling conditions (FFA concentration 0.5 mM, treatment for 24 h) and the drug concentration, the cell supernatant was discarded, cells were washed three times with PBS, and cells were collected by trypsin digestion. After centrifugation and discarding the supernatant, 150 μL of lysis buffer was added to lyse the cells. The cell lysate was collected and incubated on ice for 30 min. Intracellular triglyceride (TG) levels were measured according to the instructions of the corresponding kits. Significant changes in TG levels were observed at the cellular level. Results are as follows: Figure 3 As shown in C.

[0053] Depend on Figure 3 As can be seen from C, the TG content in the model group reached 0.57 mmol / gprot, which was significantly increased compared with the blank group; while after intervention with 60% ethanol extract of Artemisia argyi, the TG content in the treatment group decreased in a dose-dependent manner, and the TG content in the high-dose group was as low as 0.44 mmol / gprot, which was highly statistically significant compared with the model group (P<0.01).

[0054] Figure 3 The results showed that the 60% ethanol extract of Artemisia annua could inhibit lipid accumulation in a dose-dependent manner.

[0055] 5. In vivo model experiment of NAFLD induced by high-fat diet (HFD) in mice Six- to eight-week-old C57 mice were acclimatized for one week and then randomly divided into two groups: a control group and a model group. The control group was fed a low-fat, low-sugar control diet (TP 26352, Nantong Trofi Feed Technology Co., Ltd., China), while the model group was fed a high-fat, high-cholesterol diet (TP 26304, Nantong Trofi Feed Technology Co., Ltd., China; 42% fat, 0.2% cholesterol). Both diets were maintained for 12 weeks. At week 11, two mice were randomly selected for dissection to assess various indicators and determine the model establishment status. After successful modeling, mice were randomly divided according to body weight into a normal group (low-fat, low-sugar control diet), a model group (high-fat, high-cholesterol diet), a low-dose group (60% ethanol extract of Artemisia argyi, 0.5 g / kg, L-AEAR; fed a high-fat, high-cholesterol diet), a medium-dose group (60% ethanol extract of Artemisia argyi, 1 g / kg, M-AEAR; fed a high-fat, high-cholesterol diet), a high-dose group (60% ethanol extract of Artemisia argyi, 2 g / kg, H-AEAR; fed a high-fat, high-cholesterol diet), and a positive control group (ATO, atorvastatin, 10 mg / kg; fed a high-fat, high-cholesterol diet). The normal and model groups were administered sodium carboxymethyl cellulose solution (CMCNa), while the remaining groups were administered the dosage determined by subacute toxicity experiments.

[0056] In week 15, a glucose tolerance test was conducted by fasting: Before the test, mice were fasted for 6 hours but allowed free access to water. After fasting, fasting blood glucose (0 min) was measured for each mouse. Glucose (2 g / kg saline) was injected into the mice according to their body weight to conduct an intraperitoneal glucose tolerance test (OGTT). Blood glucose levels at the tip of the mice's tails were measured at 15, 30, 45, 60, and 120 min after injection.

[0057] Sample collection: After the last administration, blood was collected from the eyeballs of mice, and the mice were euthanized by cervical dislocation. The liver, spleen, kidney, epididymis, and abdominal fat were harvested and weighed, and organ indices were calculated. The liver and fat were rinsed with physiological saline and stored at -80°C for later use. The levels of four related indicators of serum liver function and blood lipids in mice were detected using a kit.

[0058] Figure 4 The results of the experiment in Example 1 on the relief of HFD-induced NAFLD by 60% ethanol extract of Artemisia argyi; where AB are the images showing the changes in mouse body weight and liver ultrasound after drug administration; C is the statistical graph of mouse body weight change after drug administration; D is the statistical graph of cumulative food intake and body weight of mice after drug administration; E is the statistical graph of mouse body weight before dissection. Figure 5The results of the experiment in Example 1 on the relief of HFD-induced NAFLD by 60% ethanol extract of Artemisia argyi; where A is a graph showing the changes in liver and epididymal fat weight in mice after administration; B is a statistical graph showing the changes in liver in mice after administration; C is the epididymal fat index in mice; D is the abdominal fat index in mice; EG is a graph showing the blood glucose test results. Figure 6 The results of the 60% ethanol extract of Artemisia argyi in Example 1 on the levels of lipid-related indicators in the serum and liver of mice induced by HFD are shown below. Among them, A represents the change in serum AST level; B represents the change in serum ALT level; C represents the change in serum LDL level; D represents the change in serum HDL level; E represents the change in serum TG level; F represents the change in serum TC level; G represents the change in liver TG level; and H represents the change in liver TC level.

[0059] The test results show that the mice underwent a significant decrease in body weight after administration. Not only were serum AST, TG, TC, AST, and ALT levels improved after administration, but liver TC and TG levels were also significantly improved.

[0060] The livers and epididymal fat of three mice from each group were immersed in paraformaldehyde. The livers were stained with Oil Red O and HE, and the epididymal fat was stained with HE. The results are as follows: Figure 7 As shown.

[0061] Figure 7 The pathological staining results of the normal group (CON), model group (FDA), low-dose group (LD), medium-dose group (MD), high-dose group (HD), and positive control group (ATO) are summarized. Among them, A is the staining results; B is the statistics of Oil Red O staining results; C is the statistics of HE staining results; and D is the HE staining results of epididymal fat.

[0062] The results showed that, compared with the model group, lipid accumulation was significantly reduced after drug administration, vacuolation and inflammatory cell infiltration were improved, collagen deposition was significantly reduced, and the area of ​​adipocytes was also significantly reduced.

[0063] The extraction process for Artemisia annua extract designed in this invention is simple and efficient, using a 60% (v / v) ethanol aqueous solution as the extraction solvent to obtain the active ingredient from Artemisia annua. In vitro and in vivo NAFLD activity studies elucidated the mechanism of action of Artemisia annua extract against NAFLD, providing a scientific basis for the rational utilization of Artemisia annua resources and laying a theoretical foundation for the research and development of Artemisia annua as a hepatoprotective and anti-NAFLD drug.

[0064] This invention studies the role of Artemisia annua in resisting NAFLD, providing a scientific basis for the rational development and utilization of medicinal resources.

[0065] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. The use of an Artemisia annua extract in the preparation of a medicament for the treatment and / or prevention of non-alcoholic fatty liver disease.

2. The application according to claim 1, characterized in that, The preparation method of the Artemisia argyi extract includes the following steps: The powdered Artemisia argyi was mixed with a 60% (v / v) ethanol aqueous solution for extraction, followed by solid-liquid separation to obtain the ethanol extract. The alcohol extract was concentrated to obtain an extract, which was then dried to obtain the Artemisia argyi extract.

3. The application according to claim 2, characterized in that, The ratio of Artemisia argyi powder to ethanol aqueous solution is 1g:15~25mL.

4. The application according to claim 2, characterized in that, The extraction time is 60-84 hours; The concentration temperature is 40~60℃ and the pressure is 0.07~1MPa.

5. The application according to claim 2, characterized in that, The drying process is freeze drying; the freeze drying temperature is -43.4℃, the vacuum degree is 4.6Pa, and the time is 6~10h.

6. The application according to claim 1, characterized in that, The medications used to treat and / or prevent non-alcoholic fatty liver disease are those that alleviate liver function and lipid abnormalities.

7. The application according to claim 1, characterized in that, The medications used to treat and / or prevent non-alcoholic fatty liver disease are those that reduce lipid deposition in the liver.

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