Composition for preventing and treating non-alcoholic steatohepatitis and preparation and application thereof

Abstract

The invention discloses a composition for preventing and treating non-alcoholic steatohepatitis and preparation and application thereof.The preparation method includes the steps that olive leaves and boxthorn leaves serve as raw materials and are extracted, concentrated and filtered, and then an olive leaf extract with the oleuropein content larger than 50% and a boxthorn leaf extract with the total flavone content larger than 60% are obtained through macroporous resin separation and purification; and mixing different olive leaf extracts and boxthorn leaf extracts according to a certain ratio to obtain the composition for preventing and treating the non-alcoholic steatohepatitis, wherein the composition has a remarkable synergistic effect in prevention of the non-alcoholic steatohepatitis.

Description

Technical Field

[0001] The present invention relates to a composition for preventing and treating hepatitis, a preparation method and application thereof, and in particular to a composition for preventing and treating non-alcoholic fatty hepatitis, a preparation method and application thereof, and belongs to the field of medical technology. Background Art

[0002] Non-alcoholic fatty liver disease refers to a clinical pathological syndrome characterized by excessive fat deposition in hepatocytes excluding alcohol and other clear liver-damaging factors. It is an acquired metabolic stress liver injury closely related to insulin resistance and genetic susceptibility. It includes simple fatty liver, non-alcoholic fatty liver disease and its related cirrhosis. Non-alcoholic fatty hepatitis is a clinical syndrome with pathological changes similar to alcoholic hepatitis but without a history of excessive drinking. It is more common in middle-aged people, especially overweight and obese individuals. Non-alcoholic fatty hepatitis is closely related to metabolic disorders such as obesity, insulin resistance, type 2 diabetes, and hyperlipidemia. Its main characteristics are macrovesicular fatty degeneration of hepatocytes accompanied by hepatocyte damage and inflammation. Severe cases can develop into cirrhosis, and there is no special treatment.

[0003] At present, Chinese medicine compositions composed of multiple Chinese medicines have been published for the prevention / treatment of non-alcoholic fatty hepatitis. For example, patent CN115998831A discloses a composition composed of 21 Chinese medicines including coptis chinensis, pinellia ternata, atractylodes lancea, peach kernel, coix seed fried with bran, citron, curcuma, and ligusticum chuanxiong for the prevention and treatment of non-alcoholic fatty hepatitis; patent CN116036190A discloses a composition composed of four raw materials including capillaries, dandelions, lilies, and lindera spatholobi, which can effectively reduce lipid deposition in the liver of mice with non-alcoholic fatty hepatitis, improve liver fibrosis, reduce inflammatory cells in damaged livers of mice, and protect liver function; the patent discloses an important composition composed of salvia miltiorrhiza, lotus leaf, gynostemma pentaphyllum, curcuma, atractylodes macrocephala, artemisia capillaris, herba scutellariae, and bupleurum for the prevention and treatment of non-alcoholic fatty hepatitis. These disclosed compositions for preventing and treating non-alcoholic fatty liver disease are all composed of multiple Chinese medicines, with too many medicinal flavors, multiple heads and multiple roots, and unclear main lines. In addition, different dosages and ratios will also lead to different efficacy. At the same time, there is no report on the combination of olive leaf and wolfberry leaf extract for preventing and treating non-alcoholic fatty liver disease. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide a composition for preventing and treating non-alcoholic fatty hepatitis, a preparation method and application thereof, in view of the deficiencies in the prior art. The present invention mixes an olive leaf extract and a wolfberry leaf extract to obtain a composition for preventing and treating non-alcoholic fatty hepatitis. The drug composition is simple, the olive leaf extract and the wolfberry leaf extract have the same or similar medicinal effects, and the two effectively compounded show significant synergistic effects in the prevention of non-alcoholic hepatitis.

[0005] In order to achieve the above object, the present invention adopts the following technical solution:

[0006] One of the purposes of the present invention is to provide a method for preparing a composition for preventing and treating non-alcoholic fatty hepatitis, comprising the following steps:

[0007] (1) Extraction of olive leaf extract and wolfberry leaf extract:

[0008] Olive leaves and wolfberry leaves are used as raw materials, olive leaves and wolfberry leaves are subjected to reflux extraction several times by using an extraction solvent, the extracts are respectively combined and concentrated to obtain concentrated solutions, and the concentrated solutions are respectively fixed to a certain concentration as loading solutions, thereby obtaining olive leaf loading solutions and wolfberry leaf loading solutions;

[0009] (2) Macroporous resin separation sample solution:

[0010] The macroporous resin is loaded into a column by a wet method, the olive leaf sample solution and the wolfberry leaf sample solution are firstly adsorbed by the macroporous resin, respectively, and then eluted by deionized water, respectively, and finally the macroporous resin is eluted by an elution solvent, respectively, the eluates are collected and concentrated under reduced pressure, respectively, and dried to obtain the olive leaf extract and the wolfberry leaf extract; the content of oleuropein in the olive leaf extract and the content of total flavonoid extract in the wolfberry leaf extract are respectively determined, and the contents meet the compounding requirements and are set aside for use;

[0011] (3) Preparation of composition:

[0012] The olive leaf extract and the wolfberry leaf extract obtained in step (2) are mixed in a certain mass ratio to obtain the composition for preventing and treating non-alcoholic fatty hepatitis.

[0013] In the above technical scheme, in step (1), the extraction solvent is an ethanol aqueous solution with a mass concentration of 10% to 90%, and the solid-liquid mass ratio of olive leaves or wolfberry leaves to the extraction solvent is 1:5 to 1:30, wherein the mass of olive leaves or wolfberry leaves is measured in g and the volume of the extraction solvent is measured in mL.

[0014] In the above technical solution, in step (1), the reflux extraction is performed several times, the number of extractions is 1 to 5 times, and the extraction time for each time is 1 to 3 hours.

[0015] In the above technical scheme, in step (1), the extracts are combined for several times and then concentrated using a rotary evaporator. The temperature during concentration is 20-80°C and the pressure is 5-30 kPa. The volume of the concentrate is 1-100 times the mass of the leaves (olive leaves or wolfberry leaves), wherein the volume of the concentrate is measured in mL and the mass of the leaves is measured in g.

[0016] In the above technical solution, in step (1), the concentrated liquid is fixed to a certain volume, and the volume after the fixed volume is 5 to 15 times the mass of the leaves, preferably 10 times; wherein the volume is measured in ml and the mass is measured in g.

[0017] In the above technical solution, in step (2), the macroporous resin is a polystyrene-based resin, including but not limited to LSA-21, D101, LX-8, AB-8, HPD-100, XDA-6, LX-38, XAD-4, etc., and its column diameter-to-height ratio is 1:5 to 1:30.

[0018] In the above technical scheme, in step (2), the loading volume of the loading liquid is 2 to 15 BV, and the flow rate is 2 to 10 BV / h; the deionized water elution volume is 5 to 30 BV, and the flow rate is 5 to 20 BV / h; the elution solvent is an ethanol aqueous solution with a mass concentration of 20% to 100%, the elution volume is 2 to 15 BV, and the flow rate is 2 to 10 BV / h.

[0019] In the above technical scheme, in step (2), the reduced pressure distillation is carried out at a distillation pressure of 2 to 50 kPa and a distillation temperature of 20 to 70°C; the drying is carried out in a vacuum drying oven at a drying temperature of 20 to 80°C and a drying pressure of 2 to 40 kPa, and the drying is carried out until the water content is less than 5%.

[0020] In the above technical solution, in step (2), the olive leaf extract has an oleuropein content of not less than 60%, preferably 65% ​​to 90%.

[0021] In the above technical solution, in step (2), the wolfberry leaf extract has a total flavonoid content of not less than 60%, preferably 70% to 95%.

[0022] In the above technical solution, in step (3), the mass ratio of the olive leaf extract and the wolfberry leaf extract is (1-10):(10-1); the mass ratio is preferably (1-4):(4-1), and more preferably 3:3.

[0023] The second object of the present invention is to provide a composition for preventing and treating non-alcoholic fatty hepatitis prepared by the above-mentioned preparation method, comprising an olive leaf extract and a wolfberry leaf extract, the mass ratio of the two being (1-10): (10-1); the olive leaf extract, wherein the content of oleuropein is not less than 60%; the wolfberry leaf extract, wherein the content of total flavonoids is not less than 60%.

[0024] The third object of the present invention is to provide an application of the composition for preventing and treating non-alcoholic fatty hepatitis in the preparation of drugs for preventing and treating non-alcoholic fatty hepatitis.

[0025] The composition for preventing and treating non-alcoholic fatty hepatitis of the present invention was evaluated for activity by animal experiments, and the animal experiments were conducted on mice with non-alcoholic hepatitis induced by MCD feed. It can be seen from the animal experiments that the composition prepared by the present invention has good efficacy in preventing and treating non-alcoholic liver disease. Therefore, the present invention prepares a composition for preventing and treating non-alcoholic liver disease from olive leaves and wolfberry leaves, improves resource utilization, and the preparation process of the method is safe, low-cost, and suitable for large-scale production. DETAILED DESCRIPTION

[0026] The specific implementation of the technical solution of the present invention is described in detail below, but the present invention is not limited to the following description:

[0027] Embodiment 1:

[0028] A composition for preventing and treating non-alcoholic fatty hepatitis is prepared by the following preparation method:

[0029] (1) Extraction of olive leaf extract and wolfberry leaf extract:

[0030] Weigh 2 kg of wolfberry leaves, add 24 L of 60% ethanol solution, reflux extract for 2 hours, pour out the extract, add 24 L of 60% ethanol solution, reflux extract for 2 hours, combine the two extracts and concentrate them using a rotary evaporator. The temperature during concentration is 50°C and the pressure is 20 kPa. After recovering the ethanol, a concentrated solution is obtained. The volume of the concentrated solution is 18 L. The extract is fixed to 20 L as the wolfberry leaf loading solution.

[0031] Weigh 2 kg of olive leaves, add 24 L of 60% ethanol solution, reflux extract for 2 h, pour out the extract, add 24 L of 60% ethanol solution, reflux extract for 2 h, combine the two extracts and concentrate them using a rotary evaporator at a temperature of 55 ° C and a pressure of 25 kPa. After recovering ethanol, a concentrated solution is obtained with a volume of 19 L. The extract is fixed to 20 L as the olive leaf loading solution.

[0032] (2) Macroporous resin separation sample solution:

[0033] 2.1 Take polystyrene-based resin D101 resin wet column, the column diameter-to-height ratio is 1:8, firstly, the olive leaf sample solution is adsorbed by macroporous resin, the volume and flow rate of the olive leaf sample solution are 10BV and 2BV / h respectively; then deionized water is used for elution, the deionized water elution volume and flow rate are 5BV and 15BV / h respectively; finally, the macroporous resin is eluted with elution solvent, the eluent is 60% ethanol solution, the elution volume and flow rate are 8BV and 3BV / h respectively, the eluate is collected and concentrated under reduced pressure, after reduced pressure distillation under 15kPa pressure and 40℃ conditions, it is placed in a drying oven with a pressure of 20kPa and 50℃ for drying until the water content is less than 5% to obtain olive leaf extract.

[0034] The content of oleuropein in the olive leaf extract in this embodiment is determined by high performance liquid chromatography external standard method: 250 mg of the sample is dissolved in 80% methanol and its concentration is 2 mg / mL, the HPLC column is an ODS C18 reverse phase column (250 mm×4.6 mm, 5 μm), and the HPLC detection conditions are: the detection wavelength is 270 nm; the injection volume of the sample solution is 20 μL, the mobile phase is 0.2% acetic acid aqueous solution and acetonitrile, the mobile phase flow rate is 1 mL / min, and the order of gradient elution is 0min 86% (volume fraction) 0.2% acetic acid aqueous solution, 14% (volume fraction) acetonitrile; 30min 73% (volume fraction) 0.2% acetic acid aqueous solution, 24% (volume fraction) acetonitrile; 35min 69% (volume fraction) 0.2% acetic acid aqueous solution, 31% (volume fraction) acetonitrile. The fraction from 25 to 27 min is oleuropein. In this embodiment, the content of oleuropein in olive leaves was measured to be 67.18%, and the yield of the extract was 12%.

[0035] 2.2 Take the polystyrene-based resin LSA-21 lipid wet column with a column diameter-to-height ratio of 1:8. First, the volume and flow rate of the wolfberry leaf loading liquid are 10BV and 3BV / h respectively, the deionized water elution volume and flow rate are 5BV and 15BV / h respectively, the eluent is 60% ethanol solution, and the elution volume and flow rate are 6BV and 3BV / h respectively. Collect the eluate, carry out reduced pressure distillation at a pressure of 15kPa and 40°C, and then place it in a drying oven at a pressure of 20kPa and 50°C to dry it until the water content is less than 5% to obtain the wolfberry leaf extract.

[0036] The content of total flavonoids in the wolfberry leaf extract in this embodiment is determined by ultraviolet spectrophotometer: 250 mg of the sample is dissolved in 80% methanol to a concentration of 2 mg / mL, a 10 mm colorimetric cup is used, the reagent blank is used as the reference solution, the absorbance is measured at the maximum absorption wavelength of 510 nm, and the total flavonoid concentration and content are calculated. In this embodiment, the total flavonoid content in the wolfberry leaf is measured to be 91.32%, and the extract yield is 8%.

[0037] (3) Preparation of composition:

[0038] The olive leaf extract and wolfberry leaf extract obtained in step (2) are mixed in a mass ratio of 1:1 (denoted as A1 composition), 1:3 (denoted as A2 composition), 3:1 (denoted as A3 composition) and 3:3 (denoted as A4 composition) to obtain a composition for preventing and treating non-alcoholic fatty hepatitis.

[0039] Embodiment 2:

[0040] A composition for preventing and treating non-alcoholic fatty hepatitis is prepared by the following preparation method:

[0041] (1) Extraction of olive leaf extract and wolfberry leaf extract:

[0042] Weigh 2 kg of wolfberry leaves, add 24 L of 70% ethanol solution, reflux extract for 3 hours, pour out the extract, add 24 L of 70% ethanol solution, reflux extract for 3 hours, combine the two extracts and concentrate them using a rotary evaporator. The temperature during concentration is 45°C and the pressure is 15 kPa. After recovering the ethanol, a concentrated solution is obtained with a volume of 19 L. The extract is fixed to 20 L as the wolfberry leaf loading solution.

[0043] Weigh 2 kg of olive leaves, add 24 L of 70% ethanol solution, reflux extract for 3 hours, pour out the extract, add 24 L of 70% ethanol solution, reflux extract for 3 hours, combine the two extracts and concentrate them using a rotary evaporator at a temperature of 60°C and a pressure of 30 kPa. After recovering the ethanol, a concentrated solution is obtained with a volume of 17 L. The extract is fixed to 20 L as the olive leaf loading solution.

[0044] (2) Macroporous resin separation sample solution:

[0045] 2.1 Take polystyrene-based resin AB-8 resin wet column, the column diameter-to-height ratio is 1:10, firstly, olive leaf sample solution is adsorbed by macroporous resin, the volume and flow rate of olive leaf sample solution are 8BV and 2BV / h respectively; then deionized water is used for elution, the deionized water elution volume and flow rate are 5BV and 10BV / h respectively; finally, the macroporous resin is eluted with elution solvent, the eluent is 60% ethanol solution, the elution volume and flow rate are 6BV and 2BV / h respectively, the eluate is collected and concentrated under reduced pressure, and after reduced pressure distillation under 15kPa pressure and 40°C, it is placed in a drying oven with a pressure of 20kPa and 50°C for drying until the water content is less than 5% to obtain olive leaf extract.

[0046] The content of oleuropein in the olive leaf extract in this embodiment is determined by high performance liquid chromatography external standard method: 500 mg of sample is dissolved in 60% methanol and its concentration is 10 mg / mL, HPLC column is ODS C18 reverse phase column (250 mm×4.6 mm, 5 μm), HPLC detection conditions are: detection wavelength is 280 nm; sample injection volume is 10 μL, mobile phase is 0.2% acetic acid aqueous solution and acetonitrile, mobile phase flow rate is 1 mL / min, gradient elution sequence is 0min 86% (volume fraction) 0.2% acetic acid aqueous solution, 14% (volume fraction) acetonitrile; 30min 73% (volume fraction) 0.2% acetic acid aqueous solution, 24% (volume fraction) acetonitrile; 35min 69% (volume fraction) 0.2% acetic acid aqueous solution, 31% (volume fraction) acetonitrile. The fraction from 25 to 27 min is oleuropein. In this embodiment, the content of oleuropein in olive leaves was measured to be 66.58%, and the yield of the extract was 12%.

[0047] 2.2 Take polystyrene-based resin XAD-4 resin for wet column packing with a column diameter-to-height ratio of 1:10. First, the volume and flow rate of the wolfberry leaf loading liquid are 8BV and 2BV / h respectively, the deionized water elution volume and flow rate are 5BV and 10BV / h respectively, the eluent is 60% ethanol solution, and the elution volume and flow rate are 6BV and 2BV / h respectively. Collect the eluate, carry out reduced pressure distillation at a pressure of 15kPa and 40°C, and then place it in a drying oven at a pressure of 20kPa and 50°C to dry it until the water content is less than 5% to obtain the wolfberry leaf extract.

[0048] The content of total flavonoids in the wolfberry leaf extract in this embodiment is determined by ultraviolet spectrophotometer: 500 mg of the sample is dissolved in 60% to 100% methanol to a concentration of 10 mg / mL, a 10 mm colorimetric cup is used, and the reagent blank is used as the reference solution. The absorbance is measured at the maximum absorption wavelength of 510 nm to calculate the total flavonoid concentration and content. In this embodiment, the total flavonoid content in the wolfberry leaf is measured to be 90.83%, and the extract yield is 8%.

[0049] (3) Preparation of composition:

[0050] The olive leaf extract and the wolfberry leaf extract obtained in step (3) are mixed in a mass ratio of 3:3 to obtain the composition for preventing and treating non-alcoholic fatty hepatitis.

[0051] Application Examples

[0052] The four compositions obtained in Example 1 were used as animal experimental drugs. 82 male Kunming mice were taken, weighing 18-22g, and were randomly divided into a control group, a model group, an A1 composition group, an A2 composition group, an A3 composition group, an A4 composition group, and a positive group (Ganbao capsule 8g / kg) according to their body weight after adaptive feeding. Except that the control group was given ordinary feed, the model group and the remaining groups were all given MCD feed for 1 month. At the same time, the drug administration group was gavaged once a day, and the administration volume was 10mL / kg. It was continuously administered for 1 month, and the control group and the model group were gavaged with equal volumes of water. After the last administration, mice were fasted but not water-restricted. After 14 hours, serum was taken to detect AST, ALT, TC, TG, HDL, and LDL. The liver was taken to weigh the wet weight and calculate the liver index.

[0053] Table 1 Effects of the composition on body weight and liver index of mice

[0054]

[0055]

[0056] Compared with the model group, *P<0.05, **P<0.01, ***P<0.001.

[0057] As shown in Table 1, compared with the control group, the body weight of mice in the model group was significantly reduced and the liver index was significantly increased. Compared with the model group, the liver index of mice in the A3 composition group and the A4 composition group was significantly reduced.

[0058] Table 2 Effects of the composition on serum liver function in mice

[0059]

[0060] Compared with the model group, *P<0.05, **P<0.01, ***P<0.001.

[0061] As shown in Table 2, compared with the control group, the ALT, AST, and TBA in the serum of the mice in the model group were significantly increased, and the liver function was significantly decreased. Compared with the model group, the A1 composition, A2 composition, A3 composition, and A4 composition can significantly reduce the ALT content in the serum of mice; each administration group can significantly reduce the AST content in the serum of mice; the A2 composition group and the A4 composition group can significantly reduce the TBA content in the serum of mice.

[0062] Table 3 Effects of the composition on blood lipids in mice

[0063]

[0064] Compared with the model group, *P<0.05, **P<0.01, ***P<0.001.

[0065] As shown in Table 3, compared with the control group, the TG, TC, HDL and LDL in the serum of the mice in the model group were significantly reduced. Compared with the model group, the A2 composition can significantly reduce the TG content in the serum of mice; the A1 composition, the A2 composition, the A3 composition and the A4 composition can significantly increase the TC content in the serum of mice; each administration group can significantly increase the HDL content in the serum of mice; the A3 composition and the A4 composition can significantly increase the LDL content in the serum of mice.

[0066] Table 4 Effects of the composition on liver lipids in mice

[0067]

[0068] Compared with the model group, *P<0.05, **P<0.01, ***P<0.001.

[0069] As shown in Table 4, compared with the control group, the TG content in the liver of the mice in the model group was significantly increased. Compared with the model group, the A2 composition, the A3 composition and the A4 composition could significantly reduce the TG content in the liver of the mice.

[0070] The above examples are only for illustrating the technical concept and technical features of the present invention, and cannot be used to limit the protection scope of the present invention. Any equivalent transformation or modification made according to the essence of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a composition for preventing and treating non-alcoholic fatty hepatitis, characterized in that: The following steps are involved: (1) Extraction of olive leaf extract and wolfberry leaf extract: Olive leaves and wolfberry leaves are used as raw materials, olive leaves and wolfberry leaves are subjected to reflux extraction several times by using an extraction solvent, the extracts are respectively combined and concentrated to obtain concentrated solutions, and the concentrated solutions are respectively fixed to a certain concentration as loading solutions, thereby obtaining olive leaf loading solutions and wolfberry leaf loading solutions; (2) Separation of sample solution using macroporous resin: The macroporous resin is loaded into a column by a wet method, the olive leaf sample solution and the wolfberry leaf sample solution are firstly adsorbed by the macroporous resin, and then eluted by deionized water, and finally the macroporous resin is eluted by an elution solvent, the eluates are collected and concentrated under reduced pressure, and dried to obtain the olive leaf extract and the wolfberry leaf extract; the content of oleuropein in the olive leaf extract and the content of total flavonoid extract in the wolfberry leaf extract are respectively determined, and the contents meet the compounding requirements and are set aside for use; (3) Preparation of composition: The olive leaf extract and the wolfberry leaf extract obtained in step (3) are mixed in a certain mass ratio to obtain the composition for preventing and treating non-alcoholic fatty hepatitis.

2. The preparation method according to claim 1, characterized in that: In step (1), the extraction solvent is an ethanol aqueous solution with a mass concentration of 10% to 90%, and the solid-liquid mass ratio of olive leaves or wolfberry leaves to the extraction solvent is 1:5 to 1:30; the reflux extraction is performed several times, the number of extractions is 1 to 5 times, and the extraction time for each time is 1 to 3 hours.

3. The preparation method according to claim 1, characterized in that: In step (1), the extracts are combined and concentrated using a rotary evaporator. The temperature during concentration is 20-80° C. and the pressure is 5-30 kPa. The volume of the concentrate is 1-100 times the mass of the leaves (olive leaves or wolfberry leaves), wherein the volume of the concentrate is measured in mL and the mass of the leaves is measured in g.

4. The preparation method according to claim 1, characterized in that: Step (1), the concentrated solution is fixed to a certain concentration, and the volume after the fixed concentration is 5 to 15 times the mass of the leaves.

5. The preparation method according to claim 1, characterized in that: In step (2), the macroporous resin is a polystyrene-based resin, specifically any one of LSA-21, D101, LX-8, AB-8, HPD-100, XDA-6, LX-38, and XAD-4, and its column diameter-to-height ratio is 1:5 to 1:

30.

6. The preparation method according to claim 1, characterized in that: In step (2), the loading volume of the loading solution is 2~15 BV, and the flow rate is 2~10 BV / h; the deionized water elution volume is 5~30 BV, and the flow rate is 5~20 BV / h; the elution solvent is an ethanol aqueous solution with a mass concentration of 20%~100%, the elution volume is 2~15 BV, and the flow rate is 2~10 BV / h.

7. The preparation method according to claim 1, characterized in that: In step (2), the reduced pressure distillation is carried out at a distillation pressure of 2 to 50 kPa and a distillation temperature of 20 to 70 °C; the drying is carried out in a vacuum drying oven at a drying temperature of 20 to 80 °C and a drying pressure of 2 to 40 kPa, and the drying is carried out until the water content is less than 5%.

8. The preparation method according to claim 1, characterized in that: In step (2), the olive leaf extract has an oleuropein content of not less than 60%; the wolfberry leaf extract has a total flavonoid content of not less than 60%; in step (3), the olive leaf extract and the wolfberry leaf extract have a mass ratio of (1-10):(10-1).

9. A composition for preventing and treating nonalcoholic fatty hepatitis prepared by the preparation method according to any one of claims 1 to 8, characterized in that: The invention comprises an olive leaf extract and a wolfberry leaf extract, wherein the mass ratio of the two is (1-10): (10-1); the olive leaf extract contains no less than 60% oleuropein; and the wolfberry leaf extract contains no less than 60% total flavonoids.

10. Use of the composition for preventing and treating non-alcoholic fatty hepatitis according to claim 9 in preparing a drug for preventing and treating non-alcoholic fatty hepatitis.

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