Application of prinsepia utilis royle seed meal extract

Abstract

The invention discloses application of a prinsepia utilis royle seed meal extract. The method comprises the following steps: by taking dried prinsepia utilis royle as a raw material, crushing the prinsepia utilis royle, adding petroleum ether, performing reflux extraction, filtering, removing an extracting solution, and volatilizing filter residues to obtain prinsepia utilis royle seed meal; adding an ethanol solution into the prinsepia utilis royle seed meal for reflux extraction, filtering to obtain an extracting solution, and concentrating the extracting solution under reduced pressure until the extracting solution is dry to obtain the prinsepia utilis royle seed meal extract. The prinsepia utilis royle seed meal extract prepared by the invention has the function of improving hepatic fibrosis, can be used as an active ingredient for preparing medicines, health-care products or foods for preventing and treating hepatic fibrosis, and provides a new way for preparing medicines and health-care products with anti-hepatic fibrosis activity.

Description

Technical Field

[0001] The invention relates to the field of medicine and health technology, and in particular to application of a prickle fruit seed meal extract. Background Art

[0002] Liver fibrosis is the liver's repair response to chronic damage (such as hepatitis, alcohol, fatty liver, etc.), which is manifested as abnormal proliferation of fibrous connective tissue in the liver and is the early stage of cirrhosis. The essence is that after the activation of hepatic stellate cells, a large amount of extracellular matrix is ​​synthesized and deposited, leading to liver structural destruction and functional impairment. At present, clinical anti-liver fibrosis drugs are still in the exploratory stage, and there are no recognized specific drugs. Drug research and development focuses on HSC activation, ECM metabolism, inflammation regulation, etc., with limited efficacy and obvious side effects. The discovery of natural products with anti-liver fibrosis activity is an urgent clinical need.

[0003] Prinia prickly pear is the fruit of a perennial deciduous shrub of the genus Psoralea in the Rosaceae family, mainly distributed in high-altitude areas such as Yunnan, Sichuan, and Tibet. Prinia prickly pear oil is mainly used in folk applications, and has pharmacological activities such as antibacterial, anti-inflammatory, antioxidant, hypolipidemic, and reduced scar formation. Patent CN202310846712.4 extracted 9 active peptides from prinia prickly pear, and found that it had significant anti-inflammatory and antioxidant effects on the acute pancreatitis model. Patent CN202410158751.X discloses a prinia prickly pear extract with skin barrier repair efficacy, its preparation method, and application. A high-purity extract is obtained by microwave treatment and alcohol precipitation process, which can significantly improve the skin barrier repair effect.

[0004] However, although many studies have focused on the medicinal value of Prinus prinus and its extracts, the medicinal value of Prinus prinus seed meal ethanol extract is still relatively scarce and urgently needs further in-depth exploration and excavation. Summary of the invention

[0005] In view of the deficiencies of the prior art, the present invention provides an application of a prickle seed meal extract. The prickle seed meal extract prepared by the present invention can be used to prepare medicines, health products or foods for preventing and treating liver fibrosis, broadens the way for the treatment of liver fibrosis, and is of great significance for the development and utilization of prickle fruit.

[0006] The technical solution of the present invention is as follows:

[0007] The invention discloses an application of a prickle seed meal extract, wherein the prickle seed meal extract is used as an active ingredient in preparing at least one of a medicine, a health product or a food for preventing and treating liver fibrosis.

[0008] Furthermore, the preparation method of the utilitarian fruit seed meal extract comprises the following steps:

[0009] (1) Defatting: Using dry Prinsepia utilis Royle fruits as raw materials, after pulverization, petroleum ether is added for reflux extraction, filtration, discarding the extract, and the filter residue is dried to obtain Prinsepia utilis Royle seed meal;

[0010] (2) Ethanol extraction: Ethanol solution is added to the Prinsepia utilis Royle seed meal for reflux extraction, filtration is carried out to obtain the extract, and the extract is concentrated to dryness under reduced pressure to obtain the Prinsepia utilis Royle seed meal extract.

[0011] Furthermore, in step (1), the mass-volume ratio of the Prinsepia utilis Royle fruits to petroleum ether is 1 g: 6 - 10 mL.

[0012] Furthermore, in step (1), the temperature of the reflux extraction is 70 - 90 °C, the time is 30 - 120 min, and the number of times is 1 - 3 times.

[0013] Furthermore, in step (2), the volume fraction of the ethanol solution is 70 - 90%.

[0014] Furthermore, in step (2), the mass ratio of the ethanol volume to the mass of the Prinsepia utilis Royle fruits in step (1) is 6 - 10 mL: 1 g.

[0015] Furthermore, in step (2), the temperature of the reflux extraction is 100 - 120 °C, the time is 30 - 120 min, and the number of times is 1 - 3 times.

[0016] The beneficial technical effects of the present invention are as follows:

[0017] The present invention further develops and utilizes the residue remaining after reflux extraction with petroleum ether, uses ethanol to carry out reflux extraction on the Prinsepia utilis Royle seed meal, and concentrates the extract to dryness to obtain the Prinsepia utilis Royle seed meal extract. The prepared Prinsepia utilis Royle seed meal extract can significantly inhibit the proliferation of LX-2 cells, significantly reduce the levels of ALT and AST in the serum of mice with carbon tetrachloride-induced liver fibrosis, improve liver fibrosis, and can be used as an active ingredient for preparing drugs, health products or foods for preventing and treating liver fibrosis, providing a new way for preparing drugs and health products with anti-liver fibrosis activity. Description of the Drawings

[0018] Figure 1 It is a graph showing the effect of the Prinsepia utilis Royle seed meal extract prepared in Example 1 of the present invention on the viability of human hepatic stellate cells.

[0019] Figure 2 It is a graph showing the effect of the Prinsepia utilis Royle seed meal extract prepared in Example 1 of the present invention on the levels of serum alanine aminotransferase and aspartate aminotransferase in mice with carbon tetrachloride-induced liver fibrosis.

[0020] Figure 3 It is a histological structure diagram of the liver of mice with liver fibrosis treated with the Prinsepia utilis Royle seed meal extract prepared in Example 1 of the present invention. Detailed implementation mode

[0021] The present invention will be specifically described below in conjunction with the accompanying drawings and embodiments.

[0022] Example 1

[0023] A preparation method of an extract from Prinsepia utilis Royle seed meal comprises the following steps:

[0024] (1) Degreasing: Take 50 g of Prinsepia utilis Royle, crush it, add 400 mL of petroleum ether, reflux and extract at 80 °C for 60 min, extract 2 times, discard the extract, and let the filter residue stand to volatilize the petroleum ether to obtain Prinsepia utilis Royle seed meal;

[0025] (2) Alcohol extraction: Add 400 mL of 80% ethanol to the Prinsepia utilis Royle seed meal, reflux and extract at 110 °C for 60 min, extract 2 times, combine the extracts, and concentrate under reduced pressure to dryness to obtain 9.1 g of an extract from Prinsepia utilis Royle seed meal, with a yield of 18.2%.

[0026] Example 2

[0027] A preparation method of an extract from Prinsepia utilis Royle seed meal comprises the following steps:

[0028] (1) Degreasing: Take 50 g of Prinsepia utilis Royle, crush it, add 300 mL of petroleum ether, reflux and extract at 70 °C for 30 min, extract 3 times, discard the extract, and let the filter residue stand to volatilize the petroleum ether to obtain Prinsepia utilis Royle seed meal;

[0029] (2) Alcohol extraction: Add 300 mL of 70% ethanol to the Prinsepia utilis Royle seed meal, reflux and extract at 120 °C for 30 min, extract 3 times, combine the extracts, and concentrate under reduced pressure to dryness to obtain 10.3 g of an extract from Prinsepia utilis Royle seed meal, with a yield of 20.6%.

[0030] Example 3

[0031] A preparation method of an extract from Prinsepia utilis Royle seed meal comprises the following steps:

[0032] (1) Degreasing: Take 50 g of Prinsepia utilis Royle, crush it, add 500 mL of petroleum ether, reflux and extract at 90 °C for 120 min, extract 1 time, discard the extract, and let the filter residue stand to volatilize the petroleum ether to obtain Prinsepia utilis Royle seed meal;

[0033] (2) Alcohol extraction: Add 500 mL of 90% ethanol to the Prinsepia utilis Royle seed meal, reflux and extract at 100 °C for 120 min, extract 1 time, concentrate the extract under reduced pressure to dryness to obtain 7.9 g of an extract from Prinsepia utilis Royle seed meal, with a yield of 15.8%.

[0034] Test Example 1 In vitro activation test

[0035] The effects of the extract of Prinsepia utilis Royle seed meal on cell viability were tested using human hepatic stellate cells (LX-2). The specific method was as follows: LX-2 cells in good growth state were made into cell suspensions, plated, and after adhering and growing for 24 h, the extract of Prinsepia utilis Royle seed meal prepared in Example 1 at different concentrations (2 / 4 / 6 / 7 / 8 / 10 mg / mL) was added to the dosing group, and an equal volume of complete medium was added to the control group. After incubating for 24 h, 10 μL of CCK-8 working solution was added to each well, and it was cultured at 37 °C for 2 h. The absorbance (A) value at 450 nm was detected by an enzyme-linked immunosorbent assay (ELISA) reader, and the cell viability was calculated. The results are as Figure 1 shown. As can be seen from Figure 1 the results, the extract of Prinsepia utilis Royle seed meal can significantly inhibit the proliferation of LX-2 cells, and the IC 50 is 4.02 mg / mL.

[0036] Test Example 2 In vivo activation test

[0037] The specific method for the in vivo activity test of the extract of Prinsepia utilis Royle seed meal was as follows: 36 male C57BL / 6 mice were selected as experimental subjects, and the mice were randomly divided into a control group (Control), a model group (Modol), a positive control group (Y), and low (D) / medium (Z) / high (G) dose groups of the extract of Prinsepia utilis Royle seed meal, with 6 mice in each group. After one week of adaptive feeding, the model group and each dosing group were intraperitoneally injected with olive oil containing 10% CCl4 (the injection dose was 5 mL / kg) to induce liver fibrosis in mice, and the control group was intraperitoneally injected with an equal volume of olive oil. It was injected twice a week for 8 consecutive weeks. From the 5th week, the positive control group was intragastrically administered with 60 mg / kg of diammonium glycyrrhizinate every day, and the low (D) / medium (Z) / high (G) dose groups of the extract of Prinsepia utilis Royle seed meal were intragastrically administered with 125 / 250 / 500 mg / kg of the extract of Prinsepia utilis Royle seed meal prepared in Example 1 (corresponding to the low / medium / high dose groups of the extract of Prinsepia utilis Royle seed meal) respectively every day. The control group and the model group were intragastrically administered with an equal volume of olive oil for 4 consecutive weeks. After the last administration at the end of the 8th week, the mice were fasted but allowed to drink water for 12 h, then the mice were sacrificed, and serum and liver were taken and stored at -80 °C for standby.

[0038] (1) Effects of the extract of Prinsepia utilis Royle seed meal on the levels of serum alanine aminotransferase and aspartate aminotransferase in carbon tetrachloride-induced liver fibrosis mice

[0039] The contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum samples were detected using a Mindray BS-420 automatic biochemical analyzer respectively. The results are as Figure 2 shown. As can be seen from Figure 2 the results, the levels of serum alanine aminotransferase and aspartate aminotransferase in the model group mice were extremely significantly higher than those in the control group, and both the positive control group and the high dose group of the extract of Prinsepia utilis Royle seed meal could significantly reduce the levels of ALT and AST in the serum of mice.

[0040] (2) Effects of the extract of Prinsepia utilis Royle seed meal on the liver tissue of carbon tetrachloride-induced liver fibrosis mice

[0041] The livers of mice in each group were made into sections. After Masson staining and Sirius red staining, the sections were placed under an Olympus BX43 microscope for histopathological examination. The results are as Figure 3 shown. As can be seen from Figure 3 the results, under Masson staining in the control group, a small amount of delicate collagen fibers could be seen around the portal area and central vein in the liver tissue. The collagen fibers were blue, and there was no obvious collagen fiber hyperplasia. Under Sirius red staining, a small amount of collagen fibers could be seen around the portal area and vein in the liver tissue. The collagen fibers were red, and there was no obvious collagen fiber hyperplasia. Under Masson staining in the model group, there was collagen fiber hyperplasia between multiple portal areas and between the portal area and the central vein. The collagen fibers were stained blue, and the proliferated collagen fibers were connected to form fibrotic bridging, showing moderate fibrosis. Under Sirius red staining, there was collagen fiber hyperplasia between multiple portal areas and between the portal area and the central vein. The collagen fibers were stained red, and the proliferated collagen fibers were connected to form fibrotic bridging, showing moderate fibrosis. There was no obvious improvement in the positive control group and the medium / low-dose groups of the extract of Prinsepia utilis Royle seed meal. Under Masson staining, there was collagen fiber hyperplasia between multiple portal areas and between the portal area and the central vein. The collagen fibers were stained blue, and the proliferated collagen fibers were connected to form fibrotic bridging, showing moderate fibrosis. Under Sirius red staining, there was collagen fiber hyperplasia between multiple portal areas and between the portal area and the central vein. The collagen fibers were stained red, and the proliferated collagen fibers were connected to form fibrotic bridging, showing moderate fibrosis. Under Masson staining in the high-dose group of the extract of Prinsepia utilis Royle seed meal, there was collagen fiber hyperplasia between a small amount of portal areas and between the portal area and the central vein. The collagen fibers were stained blue, and a small amount of proliferated collagen fibers were connected to form fibrotic bridging. There was collagen fiber hyperplasia between a small amount of portal areas and between the portal area and the central vein. The collagen fibers were stained red, and a small amount of proliferated collagen fibers were connected to form fibrotic bridging. It shows that the extract of Prinsepia utilis Royle seed meal prepared by the present invention has a significant improvement effect on carbon tetrachloride-induced liver fibrosis.

[0042] The above are only the preferred embodiments of the present invention, and the present invention is not limited to the above examples. It can be understood that other improvements and changes directly derived or associated by those skilled in the art without departing from the spirit and concept of the present invention should be considered to be included within the protection scope of the present invention.

Claims

1. Application of a Prinsepia utilis Royle seed meal extract, characterized in that, The extract of Prinsepia utilis Royle seed meal is used as an active ingredient for preparing at least one of a medicament, a health product or a food for preventing and treating liver fibrosis.

2. The application according to claim 1, wherein The preparation method of the extract of Prinsepia utilis Royle seed meal comprises the following steps: (1) Degreasing: Using dry Prinsepia utilis Royle as raw material, after pulverizing, adding petroleum ether for reflux extraction, filtering, discarding the extract, and drying the filter residue to obtain Prinsepia utilis Royle seed meal; (2) Ethanol extraction: Adding an ethanol solution to the Prinsepia utilis Royle seed meal for reflux extraction, filtering to obtain an extract, and concentrating the extract under reduced pressure to dryness to obtain the extract of Prinsepia utilis Royle seed meal.

3. The application according to claim 2, characterized in that, In step (1), the mass-volume ratio of Prinsepia utilis Royle to petroleum ether is 1 g: 6-10 mL.

4. The application according to claim 2, characterized in that, In step (1), the temperature of the reflux extraction is 70-90 °C, the time is 30-120 min, and the number of times is 1-3 times.

5. The application according to claim 2, characterized in that, In step (2), the volume fraction of the ethanol solution is 70-90%.

6. The application according to claim 2, wherein In step (2), the mass ratio of the ethanol volume to the mass of Prinsepia utilis Royle in step (1) is 6-10 mL: 1 g.

7. The application according to claim 2, characterized in that, In step (2), the temperature of the reflux extraction is 100-120 °C, the time is 30-120 min, and the number of times is 1-3 times.

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