KR102311887B1 - Composition comprising Sargassum Horner extract for preventing or treating liver disease - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prevention or treatment of liver disease, comprising the extract of hoesaengnijaban as an active ingredient. Since the effect of protecting liver cells is excellent, it can be used pharmaceutically as a composition for preventing, improving, or treating liver disease, as well as being usefully used as a health food.

Description

A pharmaceutical composition for preventing or treating liver disease, comprising an extract of hoesaengnimojaban as an active ingredient {Composition comprising Sargassum Horner extract for preventing or treating liver disease}

The present invention relates to a composition for improving, preventing, or treating liver disease, comprising an extract of hoesaengnimojaban as an active ingredient.

The liver is the organ in which metabolism occurs most actively among human body organs, and it functions to defend the whole body from external substances introduced from the outside. Since the ex vivo substance that enters the living body passes through the liver once, the liver is highly likely to be exposed to many toxic substances in addition to nutrients, and thus the probability of damage is also very high. However, the liver is an organ with excellent regenerative ability, and it recovers sufficiently to normal if there is a slight damage, but if the damage continues, a part of the liver tissue is completely destroyed and liver function is reduced. do. There are various causes of liver damage, such as generation of reactive oxygen species (ROS) due to oxidative stress, chronic stressful fatigue, excessive alcohol consumption, viral infection, and harmful substances such as various drugs. When such liver damage becomes chronic, it progresses to liver failure, liver fibrosis, liver cirrhosis, liver cancer, etc. regardless of the cause.

In the case of various types of hepatocellular damage or liver disease, there is no definite cure, so it is very important to develop a drug that can prevent or treat liver disease or liver damage itself. In particular, there is a growing need to develop high value-added natural materials that improve liver function and protect against liver damage while maintaining the structure and function of liver tissue without side effects or toxicity.

Sargassum horneri is a yellowish-brown plant belonging to the family Phaeophyceae, Fucale, and Sargassaceae. The anti-osteoporosis effect (Yakugaku Zasshi. 2006, 126, 1117-1137), the blood clotting effect (Bioresour. Technol. 2007, 98, 1711-1716), and the inhibition of Herpes simplex virus type 1 It has been reported to have an effect (Chem. Pharm. Bull. 2001, 49, 484-485.; Biol. Pharm. Bull. 1998, 21, 730-734) and atopic dermatitis treatment effect. However, nothing is known about the pharmacological effects of hoesaengi hat on liver disease.

Republic of Korea Patent Registration No. 10-1613693

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating liver disease, comprising the extract of hoesaengi capancius as an active ingredient.

Another object of the present invention is to provide a health functional food for preventing or improving liver disease, which contains the extract of hoesaengnimojaban as an active ingredient.

One aspect of the invention provides a Hoe life is Sargassum (Sargassum horneri), liver disease prevention or treatment a pharmaceutical composition containing the extract as an active ingredient.

The extract according to an embodiment of the present invention (1) H ₂ O ₂ increases the reduced hepatocyte viability, (2) inhibits the production of ROS (reactive oxygen species) in hepatocytes, (3) Inhibits the production of apoptotic bodies in hepatocytes, (4) suppresses the expression of Bax and p53 proteins, which are pro-apoptosis proteins, and Bcl-2, which is an anti-apoptosis protein It can prevent hepatocellular damage by increasing the expression of and PARP (reducing cleared PARP), so it can be usefully used for the prevention, improvement and treatment of liver disease.

The extract according to one embodiment of the present invention can prevent liver cell damage, so it is possible to fundamentally prevent or treat liver disease.

According to one embodiment of the present invention, the extract of hoesaengnimojaban is water, alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, butyl acetate, chloroform, diethyl ether, dichloromethane, hexane , and may be extracted with a solvent selected from the group consisting of mixtures thereof.

According to one embodiment of the present invention, the alcohol having 1 to 4 carbon atoms may be methanol or ethanol.

According to one embodiment of the present invention, the extract may be extracted with a solvent mixed with water and ethanol in a volume ratio of 2:8 to 4:6, or 3:7.

In one embodiment, the solvent in which water and ethanol are mixed may be 30% ethanol, 40% ethanol, 50% ethanol, 60% ethanol, 70% ethanol, 80% ethanol, or 90% ethanol, preferably 70 % ethanol. Extraction of black serrata with a solvent containing ethanol may increase the content of components useful for hepatocellular protection. The present inventors confirmed that the polyphenol content of the extract was significantly increased when extracting hoesaengi hatban with ethanol as a solvent compared to extracting with methanol as a solvent.

In one embodiment, the hoesaengi hat extract may be extracted at 60 to 80 ℃, 65 to 75 ℃, or 70 ℃. The hoesaengi hat extract may have a ratio of major components depending on the extraction temperature, and extraction at the temperature may increase the content of components that help protect hepatocytes. The present inventors confirmed that, when extracting hoesaengi capbanol under heating conditions with ethanol as a solvent, the content of mosabanchromanol contained in the extract significantly increased than when extracting under room temperature conditions.

In one embodiment, the ethanol extract may be extracted for 8 to 14 hours, 10 to 14 hours, or 12 hours. In the ethanol extract, the ratio of the main components may be changed according to the extraction time, and when the extraction time is performed, the content of the components helpful in hepatocellular protection may be increased.

In the present invention, "extract" refers to an extract obtained by extracting herbal medicines, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a prepared or purified product of the extract, or a mixture thereof, and the extract itself and the extract. It includes extracts of all formulations that can be formed by

According to one embodiment of the present invention, the extract may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze-drying or spray-drying.

According to one embodiment of the present invention, the extract of hoesaengnimojaban includes an extract that has undergone a conventional purification process. For example, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity, or affinity), etc. Fractions obtained through various purification methods may also be included in the extract of the present invention.

The composition according to one embodiment of the present invention may include 0.001% by weight to 99.99% by weight, preferably 0.1% to 99% by weight of the extract based on the total weight of the composition, which prevents liver disease or According to the method and purpose of use of the therapeutic composition, it may be appropriately selected by those of ordinary skill in the art to which the present invention pertains.

According to one embodiment of the present invention, liver disease may be due to oxidative stress.

According to one embodiment of the present invention, the composition comprising the extract of hoesaengi capricorn may be to protect hepatocytes from oxidative stress.

According to one embodiment of the present invention, the composition comprising the extract of hoesaengi capricorn may be to inhibit hepatocellular death due to oxidative stress.

Liver tissue is subjected to oxidative damage by various factors of oxidative stress (heavy metals, oxidizing agents, alkylating agents, etc.) (Vitaglione et al., 2004, Crit. Rev. Food Sci. Nutr. 44, 575-586). When hepatocellular damage caused by oxidative stress becomes chronic, various types of liver disease can develop regardless of the cause. Therefore, administration of effective antioxidants from an early stage to prevent oxidative stress in liver tissue can be a fundamental method for the prevention or treatment of various related liver diseases caused by oxidative stress.

According to one embodiment of the present invention, liver disease may be any one or more selected from the group consisting of impaired liver function, liver failure, hepatitis, hepatotoxicity, liver fibrosis, cirrhosis and liver cancer.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive includes starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate. , lactose, mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, Calcium stearate, sucrose, dextrose, sorbitol and talc and the like can be used.

The pharmaceutical composition according to one embodiment of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition according to an embodiment of the present invention are commonly used in the preparation of drugs, and include lactose, dextrose, sucrose, sorbitol, mannitol, trehalose, hyaluronic acid, Starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. The pharmaceutical composition according to an embodiment of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (22th ed., 2013).

The pharmaceutical composition of the present invention may be administered in various oral and parenteral formulations during actual clinical administration, and in the case of formulation, commonly used fillers, extenders, binders, wetting agents, disintegrants, diluents or excipients such as surfactants It can be prepared using Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient, for example, starch, calcium, in the lipid-soluble polyphenol component-increased herbal composition of the present invention. It may be prepared by mixing carbonate (Calciumcarbonate), sucrose (Sucrose), lactose (Lactose) or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used.

The pharmaceutical composition of the present invention may be administered orally or parenterally according to a desired method. For parenteral administration, intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection injection method are used. You can choose. The dosage varies according to the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and severity of disease.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the type, severity, drug activity, and drug of the patient. Sensitivity, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs, and other factors well known in the medical field may be determined. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the composition according to one embodiment of the present invention may vary depending on the age, sex, and weight of the patient, and generally 0.1 mg to 200 mg per 1 kg of body weight, preferably 1 mg to 10 mg daily Alternatively, it may be administered every other day or divided into 1 to 3 times a day. However, since it may increase or decrease depending on the route of administration, the severity of the disease, sex, weight, age, etc., the dosage is not intended to limit the scope of the present invention in any way.

The pharmaceutical composition according to one embodiment of the present invention may be administered together with a substance exhibiting activity in one or more liver diseases.

The pharmaceutical composition according to one embodiment of the present invention may be used alone or in combination with methods using, for example, surgery, drug therapy and/or biological response modifiers for the treatment of liver disease.

Another aspect of the present invention provides a health functional food for preventing or improving liver disease, comprising the extract of hoesaengi capancius as an active ingredient.

According to one embodiment of the present invention, the health functional food contains various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, it may contain natural fruit juice and pulp for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination. In addition, health functional food is meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcoholic beverage and any one form of vitamin complex. can be

According to one embodiment of the present invention, health functional food may additionally contain food additives, and whether or not it is suitable as a "food additive" is general rules and general tests of food additives approved by the Food and Drug Administration, unless otherwise specified. Judgment is made according to the standards and standards for the relevant item in accordance with laws, etc.

According to one embodiment of the present invention, as the items described in the "Food Additives Code", for example, chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, cinnamic acid, dark pigment, licorice extract, crystalline cellulose, high-cold pigment , natural additives such as guar gum, sodium L-glutamate preparations, noodle-added alkalis, preservatives, and mixed preparations such as tar dye preparations.

The extract according to an embodiment of the present invention prevents the generation of ROS and apoptosis of hepatocytes due to oxidative stress, and thus has an excellent effect of protecting hepatocytes, so a composition for preventing, improving, or treating liver disease It can be used pharmaceutically as well as usefully as a health food.

1 shows the effect of the concentration of SE on the cell viability (%) of hepatocytes reduced by oxidative stress induced by _{H 2} O _{2 .}
Figure 2 shows the effect according to the concentration of SE on the generation of reactive oxygen species (ROS) induced by _{H 2} O _{2 .}
Figure 3 shows the results of hoechst 33342 staining showing the effect according to the concentration of SE on apoptosis production induced by _{H 2} O _{2 .}
Figure 4 shows the results of performing Western blot for the expression of apoptosis-related proteins Bcl-2, cleaved PARP, p53, and Bax when _{H 2} O _{2 and SE at various concentrations were treated in hepatocytes.}
Figure 5 shows the difference in the components of extraction at warm temperature and extraction at room temperature in the extraction of hoesaengi hat alcohol according to an embodiment.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example 1. Preparation of hoesaengi hatban extract

The hoesaengi hatban ( Sargassum horneri ) used in this example was collected from the coast of Jeju Island. 20 g of dried sagebrush hat was extracted with 70% (v/v) ethanol at 70° C. for 12 hours, filtered through filter paper (Whatman No. 6), and the resulting filtrate was concentrated and freeze-dried, and used in the experiment. Hereinafter, Sargassum horneri ethanol extract was named "SE".

Example 2. Cytotoxicity evaluation

MTT (thiazolyl blue tetrazolium bromide) analysis was performed using Chang cell (hepatocyte), which is a hepatocyte cell line, to evaluate the cytotoxicity of the extract. Chang cells (hepatocytes hereinafter) are human normal mammary epithelial cells and were purchased from the Korea Cell Line Bank.

^{Hepatocytes (1Х10 4} cells/well) were dispensed with 180 μl of DMEM medium containing 10% FBS, 1% streptomycin, and penicillin into each well of a 96-well plate, and the ethanol extract obtained from Example 1 (SE) was treated with each concentration (0 to 0.5 mg/ml). 37° C., 5% CO ₂ , After 24 hours of incubation in an incubator with saturated humidity, 15 μl of MTT solution (5 mg/ml) was added to each well and reacted for 4 hours. Then, 100 μl of a solubilization buffer containing 10% sodium dodecyl sulfate (SDS) and 50% dimethylformamide (DMSO) was added and ^{absorbance at 540 nm using a microplate reader (SpectraMax ®} M2/M2e, CA, USA). was measured. The relative cell viability in each sample treatment group was calculated by Equation 1 below.

[Equation 1]

Cell viability (%) = (S _abs /C _abs )/C _abs x 100

S _abs : absorbance of the sample and / or H ₂ O _{2 treated cells}

C _abs : absorbance of control

As a result of treating hepatocytes with SE, it was confirmed that there was no cytotoxicity at all concentrations (0-0.5 mg/ml), and the cell viability increased in a concentration-dependent manner.

Example 3. Hepatocytoprotective effect of oleracea extract against oxidative stress.

Example 3-1. Effect of H. oleracea extract on the survival rate of hepatocytes

In order to examine the effect of the extract on oxidative stress, the effect of the extract on the survival rate of hepatocytes reduced by oxidative stress induced by _{H 2} O _{2 (hydrogen peroxide) in hepatocytes was investigated through MTT analysis.} measured.

^{Hepatocytes (1Х10 4} cells/well) were dispensed with 180 μl of DMEM medium containing 10% FBS, 1% streptomycin, and penicillin into each well of a 96-well plate, and the ethanol extract obtained from Example 1 (SE) was treated for 1 hour at each concentration (0 to 0.5 mg/ml). After stimulation of hepatocytes by treatment with 1 mM H ₂ 0 _{2 , after 24 hours incubation in an incubator at 37° C., 5% CO 2} and saturated humidity, 15 μl of MTT solution (5 mg/ml) was added to each well and 4 reacted for an hour. 100 μl of a solubilization buffer containing 10% SDS and 50% DMSO was added to measure the absorbance at 570 nm using a microplate reader, and by Equation 1 above, each concentration of the sample and / or H ₂ O _{2 In the} treatment group The relative cell viability was calculated and shown in FIG. 1 .

As shown in Figure 1, when the hepatocytes were _{treated with H 2} 0 ₂ , the cell viability decreased to about 46% compared to the sample untreated control group, but when the H ₂ 0 It was confirmed that the cell viability decreased by _{2 was significantly increased.} From this result, it can be seen that the extract of hoesaengnimojapan protects hepatocytes from oxidative stress by improving the cell viability reduced by _{H 2} 0 _{2 .}

Example 3-2. Effect of hoesaengni capan extract on the generation of reactive oxygen species

It is known that when H ₂ O ₂ is treated in cells, reactive oxygen species (ROS) is generated in the cells, and the generated ROS induces oxidative stress to cause liver damage.

DCF-DA (2'-7'-dichlorofluorescin diacetate) analysis was performed to measure the amount of intracellular reactive oxygen species (ROS) production in H ₂ O _{2 treated hepatocytes.} The DCF-DA reagent enters the cell, is oxidized by oxygen radicals, is deacetylated to DCF, and converted into a fluorescent substance. At this time, the degree of oxidation can be determined by measuring the fluorescence.

^{Hepatocytes (1Х10 4} cells/well) were dispensed with 180 μl of DMEM medium containing 10% FBS, 1% streptomycin and penicillin into each well of a 96-well plate, and SE was treated for 1 hour at each concentration. After treatment with 1 mM H ₂ 0 ₂ and incubated for 1 hour in a cell incubator, 0.5 mg/ml of DCF-DA solution was added and further incubated for 5 minutes. By measuring the absorbance at 485 nm (excitation wavelength) and 528 nm (emission wavelength) using a microplate reader, the amount of ROS generation in the cell was calculated, and the results are shown in FIG. 2 . The amount of ROS production in cells was calculated by Equation 2 below.

[Equation 2]

Intracellular ROS production (%) = (S _abs /H _abs )/H _abs x 100

S _abs : absorbance of the sample and H ₂ O _{2 treated cells}

H _abs : Absorbance of cells treated with _{H 2} O _{2 only}

As a result of evaluating the inhibitory effect on intracellular ROS generation of SE by treating hepatocytes with H ₂ O _{2 , when cells were treated with H 2} O ₂ , the control group showed a significant increase in intracellular ROS, When SE was treated, the _{intracellular ROS generated by H 2} O ₂ was reduced in a concentration-dependent manner.

From the results of Examples 3-1 and 3-2, it was confirmed that the cell viability was improved by reducing the ROS in hepatocytes generated _{by H 2} O _{2 by the addition of SE.}

Example 4. Inhibitory effect of apoptosis (apoptosis) of the extract

ROS induces cell damage and apoptosis by inducing cytotoxicity by damaging cell DNA. Apoptosis is a process of phagocytosis by reducing the specific gravity of cells, destroying cell membranes, and condensing chromosomes, as well as substances inside cells to form cysts called apoptotic bodies.

During the treatment of the A. K. extract, the apoptosis inhibitory effect of the _{H. 2} O _{2 induced apoptosis formation and apoptosis was confirmed in hepatocytes.}

Example 4-1. Inhibitory effect of apoptotic body generation

Hoechst 33342 staining was performed to confirm the SE inhibitory effect on H ₂ O _{2 induced apoptosis in hepatocytes.} ^{Hepatocytes (1Х10 4} cells/well) were dispensed with 180 μl of DMEM medium containing 10% FBS, 1% streptomycin, and penicillin into each well of a 96-well plate, and SE was administered at each concentration (0 to 0.5 mg/well). ml) for 1 hour. After treatment with 1 mM H ₂ O ₂ for 12 hours, the cells were washed twice with PBS and stained with hoechst 33342 reagent (2 μg/ml) for 30 minutes using a fluorescence microscope. Confirmed.

Figure 3 shows the results of hoechst 33342 staining showing the effect according to the concentration of SE on the production of apoptosis induced by _{H 2} O _{2 .} As shown in FIG. 3, as a result of performing hoechst 33342 staining to confirm the inhibitory effect of SE on apoptosis generated by _{H 2} O _{2 treatment in hepatocytes, in the control group not treated with anything, nuclear damage} While the shape was constant without H ₂ O ₂ , it was confirmed that the nucleus was damaged and apoptosis was generated in the group treated with H 2 O 2 . However, in the case of cells treated with SE by concentration, it was confirmed that the generation of apoptosis was inhibited compared to the group treated with _{H 2} O _{2 .} From this result, it was found that SE protects hepatocytes from oxidative damage by suppressing the generation of apoptosis induced by _{H 2} O _{2 .}

Example 4-2. The effect of inhibiting the expression of apoptosis-related protein of the extract

In H ₂ O ₂ treated hepatocytes, apoptosis-related proteins Bcl-2 (B-cell lymphoma 2), cleaved PARP (poly(ADP ribose) polymers), p53 (Tumor protein p53) and Bax (Bcl- 2 Western blot was performed to examine the effect of SE on the expression of Antagonist X)).

The 10% FBS and 1% streptomycin and penicillin are included the liver (1Х10 ⁶ cells) the frequency division, and SE with DMEM medium in 10cm dishes with each concentration (0 to 0.5mg / ㎖) was treated for 1 hour . Hepatocytes were stimulated for 12 hours by treatment with 1 mM H ₂ 0 _{2 , and washed twice with PBS.} Cytoplasmic proteins were extracted from the cells using RIPA buffer (Sigma-Aldrich Co. LLC) and the protein content was quantified. Cytoplasmic proteins (30㎍) were separated by molecular weight using 10% SDS PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). The separated protein was transferred to a nitrocellulose membrane and reacted for 2 hours and 30 minutes at room temperature using 5% skim milk to prevent non-specific reaction. Antibodies against cleaved PARP, Bcl-2, p53 and Bax were mixed with 5% skim milk in a ratio of 1000:1 and reacted overnight in a refrigerated state. It was reacted with anti-mouse IgG conjugated HRP (3000:1) at room temperature for 1 hour and 30 minutes. After washing the nitrocellulose membrane 3 times with TBS-T, the expression level of the protein was checked in a Western imaging system (Davinch-K) using SuperSignal West Femto Maximum Sensitivity Substrate reagents (Thermo, USA). Figure 4 shows the results of performing Western blot for the expression of apoptosis-related proteins Bcl-2, cleaved PARP, p53, and Bax when _{H 2} O _{2 and SE at various concentrations were treated in hepatocytes.}

As shown in Figure 4, the _{cells treated with H 2} 0 ₂ compared to cells not treated with anything anti-apoptosis (anti-apoptosis) protein, Bcl-2, decreased and the expression of PARP was decreased (PARP of It was confirmed that the expression of the inactivated form of cleaved PARP increased), the pro-apoptosis proteins Bax and p53 were increased. Conversely, the SE-treated group increased the expression of Bcl-2 and PARP protein, which was decreased by _{H 2} O ₂ treatment, in a concentration-dependent manner, and the protein expression of Bax and p53 increased by _{H 2} O _{2 treatment was concentration-dependently increased.} decreased. From this result, it was confirmed that SE regulates apoptosis-related protein expression, protecting cells from oxidative damage induced by _{H 2} 0 _{2 .}

Example 5. Component analysis of ethanol extract and methanol extract

A sample of the dried serrata was extracted with 70% ethanol as a solvent at room temperature for 24 hours, and the extract was centrifuged and filtered through filter paper (Whatman No. 6) (hereinafter referred to as an ethanol-warmed extract). The contents of the extracts of the ethanol warm extract of hoesaengnijaban of Example 1 and the ethanol extract of hoesaengnimojaban of Example 5 were compared.

Referring to FIG. 5 , a difference in components between the ethanol-warmed extract and the ethanol-warmed extract was confirmed. As a result of the analysis, the ethanol-warmed extract had a high content of Mosaban chromanol, but the peak of Mosaban chromanol was not detected in the ethanol-room extract. It was confirmed that the effect of reducing ROS and increasing PARP expression in hepatocytes was more excellent in the case of extracting the locust seedling under the heating condition than the case of extracting it at room temperature. Therefore, it could be confirmed that the optimal extraction conditions to increase the content of hepatocellular protection components of the extracts of hoesaengi capricornis extracts were warm extraction with ethanol as a solvent.

Statistical processing

The experimental results of Examples 2 to 4 were evaluated for statistical significance using PASW Statistics 19.0 software (SPSS, Chicago, IL, USA). ) was used for comparison.

Claims (7)

A pharmaceutical composition for preventing or treating liver disease, comprising an extract of Sargassum horneri as an active ingredient, and the extract of hoesaengi hat is extracted at 70° C. with 70% (v/v) ethanol solvent. delete delete The method of claim 1, wherein the extract is extracted for 8 to 14 hours,
pharmaceutical composition. The pharmaceutical composition according to claim 1, wherein the liver disease is due to oxidative stress. The pharmaceutical composition according to claim 1, wherein the liver disease is at least one selected from the group consisting of liver function impairment, liver failure, hepatitis, hepatotoxicity, liver fibrosis, cirrhosis, and liver cancer. A health functional food for preventing or improving liver disease, comprising an extract of hoesaengnijaban as an active ingredient, and the extract of hoesaengnimojaban is extracted at 70°C with a 70% (v/v) ethanol solvent.

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