KR101856448B1 - Composition comprising silkworm having silk protein for preventing or treating nonalcoholic hepatitis - Google Patents

Abstract

The present invention relates to a composition containing silkworm having silk proteins. The composition containing the silkworm having silk proteins inhibits production of inflammatory substances produced by a hepatitis causing material and has an excellent liver protection effect due to inhibition of liver damaging material production, thereby being easily used for a composition for preventing or treating nonalcoholic hepatitis or as a health functional food for alleviating nonalcoholic hepatitis.

Description

Technical Field [0001] The present invention relates to a composition for preventing or treating non-alcoholic hepatitis containing silkworm silkworm protein,

The present invention relates to a composition for preventing or treating nonalcoholic hepatitis containing silkworm silk protein. More specifically, the present invention relates to a method for preventing or treating non-alcoholic hepatitis containing silkworm silkworm protein obtained by boiling a raw silkworm selected from a silkworm selected from a silkworm silkworm from a silkworm silkworm starting from the 3rd day after 5th day, ≪ / RTI >

The silkworm industry, which has produced cocoon silkworms, has been a traditional industry in Korea since the Gojoseon era. In the 1970s, more than 1 million silkworms were raised in farmhouses per year. Therefore, the current silkworm industry has turned into an industry that produces raw materials for health functional foods such as dried silkworm, sperm, and caterpillar fungus. The reason for using the 5-day-old 3-day dried silkworm is that the silkworm protein having a high strength is rapidly increased from 5 days to 3 days and from 5 days to 4 days, so that it becomes very difficult to use for food. Silk proteins are stored in silkworms in the form of silkworms, and when the silkworms become grown-up silkworms, they spit out silkworm proteins and build silkworm cocoons.

The silk which has been used for such an edible period has not been particularly used except for being used for the production of silk. However, the inventors of the present invention have found that, for the first time in Korean Patent No. 10-1388455 and Korean Patent No. 10-1546446, The silkworm silk protein silkworm has been developed as a cosmetic composition for skin improvement by pressing the silkworm silkworm at 80 ~ 125 ° C under pressure or normal pressure. However, research on silk peptide is still mainly carried out in studies on silkworm related drugs, and research using silkworm silk containing silkworm protein is insufficient.

Liver is a very important organ in charge of metabolism, detoxification, degradation, synthesis and secretion in our body. First, the liver has the function of managing the energy metabolism, and metabolizes all the nutrients absorbed from food into a substance capable of producing energy, and supplies or stores the whole body. Second, the liver has a function of synthesizing, storing and distributing fat of about 2,000 kinds of enzymes, albumin, serum proteins of bile coagulation factors, bile acid, phospholipid and cholesterol. Third, the liver has a function to excrete various metabolites through the bile duct into the duodenum, and it plays an important role in maintaining our life because of its immune function. Finally, the liver has detoxification and decomposition functions to detoxify drugs, toxic substances and alcohol. However, the hepatocyte detoxification function of this liver is likely to damage the drug, toxic, non-alcoholic liver disease can cause such.

On the other hand, the inventors of the present invention have studied the various physiological activities of cooked noodles disclosed in Korean Patent No. 10-1388455 and Korean Patent No. 10-1546446, and found that the above-mentioned noodle powder has a therapeutic effect on non-alcoholic hepatitis The present invention has been completed.

The silkworm silkworm silkworm silkworm silkworm silk protein silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm Is useful as a therapeutic agent capable of treating hepatitis using a silkworm having a silkworm.

Korean Patent No. 10-1373940 (Title of the Invention: Process for producing silkworm fermentation enzyme hydrolyzate with increased amount of physiologically active substance, hydrolyzate obtained therefrom, and use of anti-inflammatory activity of such hydrolyzate, Applicant: Chang Hee Ko et al. In, registration day: 2014.03.06) Korean Patent Laid-Open No. 10-2008-0043062 (title of the invention: anti-inflammatory agent containing silkworm fluid, applicant: Hanson Biotech Co., Ltd., registered on May 16, 2008) Korean Patent No. 10-1388455 (Title of the invention: Processing method of silkworm silkworm protein and silkworm silkworm product, Applicant: Korea (Rural Development Administration) Korean Patent No. 10-1546446 (title of the invention: health functional food and cosmetic composition comprising snack protein-containing silkworm processed product, applicant: Republic of Korea (Rural Development Administration), registration date: 2015.08.17)

It is an object of the present invention to provide a composition for preventing or treating nonalcoholic hepatitis containing silkworm silkworm containing silkworm. More specifically, the present invention relates to a method for preventing or treating non-alcoholic hepatitis containing silkworm silkworm protein obtained by boiling a raw silkworm selected from a silkworm selected from a silkworm silkworm from a silkworm silkworm starting from the 3rd day after 5th day, To provide a composition.

The present invention relates to a composition for preventing or treating nonalcoholic hepatitis containing silkworm silk protein.

The hepatitis may be a disease selected from acute hepatitis or chronic hepatitis.

The silkworm-bearing silkworm is preferably selected from silkworms ranging from four silkworms to submerged silkworms after three to five days of age.

In addition, silkworms having silkworm proteins can be obtained by boiling silkworms in water or by boiling in steam.

Drying the silkworm having the silk protein protein, and drying the silkworm silkworm through a selected one of hot air drying and vacuum freeze drying.

The silkworm-bearing silkworms may be produced in the form of an extract. The extract may be prepared by extracting silkworms having silk protein with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

The present invention provides a health functional food for preventing or ameliorating hepatitis containing silkworm silk protein.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition for preventing or treating nonalcoholic hepatitis containing silkworm silk protein. The silkworm-bearing silkworm silkworm refers to all silkworm silkworm proteins including three silkworms, four silkworms, Namely, the silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silk silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm At this time, preferably, silkworms of 5 days and 3 days and silkworms of 5 days and 4 days are preferable, and silkworm silkworms are most preferably used. It is possible to use silkworms of 5th to 3rd days, but silkworms of silkworms are smaller than silkworms of 4th and 5th days.

In addition, silkworms having silkworm proteins can be obtained by boiling silkworms in water or by boiling in steam.

On the other hand, a silkworm silkworm means silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silk silkworm The silkworm silkworm is most suitable for use as a silkworm silkworm silkworm because silkworm larvae are mostly released. On the other hand, young silkworms before 5 days of age 3 are processed as medicinal materials mainly containing silkworm, but they have a blood-sugar lowering effect and are used as diabetic drugs.

The live silkworm may be stored in a refrigerator at 5 to 10 ° C. The time to learn the silkworm is not particularly limited, but it is preferably from 55 to 125 ° C for 30 minutes to 24 hours, preferably from 80 to 104 ° C for 90 minutes to 15 hours have. The process of learning the silkworm may be repeated one to three times.

The silkworm silk protein silkworm can be dried by a conventional method after being kneaded, and further dried and pulverized by any one of natural drying, hot air drying and vacuum freeze drying. The vacuum freeze drying method is most suitable. The silkworm-bearing silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silk silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm silkworm

The silkworm-bearing silkworm silkworm can be produced in the form of an extract. In this case, the silk protein can be produced in the form of an extract, in which the raw silkworm is woven in water or in the form of steam, or the raw silkworm before being ripened can be produced in the form of an extract. It is more preferable to extract the useful component of the silkworm silkworm protein by preferably kneading the raw silkworm in water or by boiling it with steam.

In addition, the extract may be further processed into a dry powder, gel or liquid form.

The extract may be prepared by extracting a silkworm having a silk protein with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The lower alcohol having 1 to 4 carbon atoms may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol. The water, the lower alcohol having 1 to 4 carbon atoms, or the mixed solution thereof may be used at a volume of 1 to 40 times (1 to 40 l based on 1 kg), preferably 5 to 40 times the volume of the silkworm silkworm- Can be used. The extraction conditions of the extract may be from 20 to 100 ° C for 1 minute to 48 hours. The above process can be repeated 1 to 4 times. In addition, as a conventional method in the art, water of silkworm having a silk protein, a lower alcohol having 1 to 4 carbon atoms or a mixed solution extract thereof is dissolved in water, and then n-hexane, methylene chloride, acetone, chloroform, ethyl acetate and n -Butanol. The fraction may be further fractionated by using at least one solvent selected from the group consisting of ethanol, ethanol, ethanol, and butanol.

As another method, the silkworm silkworm is suspended in water, the lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof and the resultant suspension is diluted with water, 1,000-fold, more preferably 1 to 500-fold, most preferably 1 to 50-fold of water, and then adding a solvent selected from the group consisting of hexane, chloroform, ethyl acetate, and butanol to the suspension And the like.

The production temperature of the extract or its fractions may be 20 to 100 캜, but is not limited thereto. The extraction or fractionation time is not particularly limited, but it is preferable to perform extraction within 10 minutes to 2 days. As the extraction apparatus, a conventional extraction apparatus, an ultrasonic pulverization extractor, or a fractionator may be used. The extract or fraction thus prepared can be removed by hot air drying, vacuum drying or freeze drying. The extract or fraction may be purified by using column chromatography. The chromatography can be carried out using silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

In addition, the present invention can provide a pharmaceutical composition for preventing or treating nonalcoholic hepatitis containing silkworm silk protein. The pharmaceutical composition for preventing or treating nonalcoholic hepatitis containing silkworm silk protein may further comprise an appropriate carrier, excipient or diluent according to a conventional method. Examples of carriers, excipients and diluents that can be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, such as starch, calcium carbonate, sucrose, Lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used. The silkworm silk protein may be included in the composition of the present invention in an amount of 0.1 to 100% by weight.

The dosage of the composition of the present invention will vary depending on the age, sex, body weight of the subject to be treated, the particular disease or condition to be treated, the severity of the disease or condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The composition of the present invention can be administered to mammals such as rats, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. Since the composition of the present invention has little toxicity and side effects, it can be safely used for prolonged use for preventive purposes.

The present invention also provides a health functional food for preventing or ameliorating hepatitis, comprising the composition, preferably a silkworm silk protein and a pharmaceutically acceptable food additive. The silkworm silk protein may be added to the health functional food of the present invention in an amount of 0.001 to 100% by weight. The health functional food of the present invention includes forms such as tablets, capsules, pills, and liquids. Examples of the foods to which the extract of the present invention can be added include various kinds of drinks, meat, sausage, bread, Snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gums, tea and vitamin complexes.

The present invention relates to a composition for preventing or treating nonalcoholic hepatitis containing silkworm protein comprising silk protein, wherein the silkworm-containing silkworm-containing pharmaceutical composition or the health functional food is produced by the production of an inflammatory substance And thus can be easily used as a composition for preventing or treating nonalcoholic hepatitis or as a health functional food containing the same. Although a variety of health functional foods using bamboo shoots, silkworms, or silk peptides have been widely used, it is clear that the components of silk peptides differ from those of silkworms. It is confirmed that the content of each component is different when it is frozen. Bacillus subtilis is also different in growth period of silkworms and hyphae cultured, so that compared to the silkworm silkworms with different components of pharmacological activity can be said to be different. Therefore, the present invention is significant in that silkworms having silk protein, which have not been used for a long time except for breeding to make cocoon silkworm production, can be used as a composition for preventing or treating nonalcoholic hepatitis .

In the photographs and graphs in the drawings of the present invention, numerical values of 0.1, 1, and 10 indicate that silkworm powder having silk protein prepared in Example 1 was administered, and DEN indicates that diethylnitrosamine was administered.
FIG. 1 is a graph showing the total weight change, the change in liver weight change, and the total liver weight obtained after 18 hours of diethylnitrosamine (DEN) administration in SD rats fed silkworm powder of silk protein of Example 1 for 14 days Photograph, weight, and liver weight (performed in Example 3).
Fig. 2 is a photograph showing hepatoxylin & eosin staining of liver tissue obtained in Example 4. Fig.
FIG. 3 shows the results of ALP (alkaline phosphatase), ALT (alanine transaminase) and AST (aspartate transaminase) concentrations in blood of SD rats euthanized in Example 4. FIG.
FIG. 4 shows Western blot results of protein levels of COX-2 and iNOS in liver tissue of euthanized SD rats.
FIG. 5 shows the results of confirming the expression levels of F4 / 80 and CD31 in liver tissue of euthanized SD rats.
FIG. 6 shows the results of confirming the expression level of T cell activation factor in liver tissue of euthanized SD rats.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art.

Example 1: Preparation of silkworm powder (cooked silkworm powder) having silk protein>

Silkworm silkworms (White-jade, Golden-silk, Light-green 3 varieties) were prepared as silkworms with silk protein. For this purpose, the silkworms which reached the submergence around 5th and 7th day of age were boiled for 10 hours with 100 ℃ water vapor, then they were cooled and vacuum lyophilized. The dried chopped silk was crushed with a hammer mill and a roller mill into a powder having a diameter of 0.1 mm or less to prepare a final silkworm powder. At this time, as a comparative group for compositional analysis, quick-freeze-freeze-dried freeze-dried powder was prepared.

≪ Example 2 &

The main components of the experiment were as follows (Table 1 to Table 4).

division Take a nap before learning Snooze moisture 4.15 2.85 Crude protein 62.43 68.66 Crude fat 11.66 10.99 Views min 3.63 3.26 Crude fiber 1.74 1.77

division Take a nap before learning Snooze Cysteine (CYS) 0.463 0.427 Methionine (MET) 0.756 0.644 Aspartic acid (ASP) 4.850 4.679 Threonine (THR) 2.265 2.185 Serine (SER) 6.194 6.757 Glutamic acid (GLU) 4.071 3.824 Glycine (GLY) 9.265 12.335 Alanine (ALA) 7.460 9.778 Valine (VAL) 2.292 2.399 Isoleucine (IIe) 1.181 1.214 Leu 2.114 1.983 Tyrosine (Tyr) 3.901 4.560 Phenylalanine (Phe) 1.740 1.746 Lysine (Lys) 2.396 2.193 Histidine (His) 1.160 1.083 Arginine (Arg) 2.123 1.958 Proline (Pro) 1.217 1.148 Tryptophan (Trp) 0.437 0.425

division Take a nap before learning Snooze Myristicacid (C14: 0) 0.15 0.17 Palmiticacid (C16: 0) 23.98 24.49 Palmitoleicacid (C16: 1? 7) 0.82 0.86 Stearicacid (C18: 0) 8.37 8.55 Oleicacid (C18: 1? 9) 31.13 31.22 Vaccenicacid (C18: 1? 7) 0.00 0.00 Linoleicacid (C18: 2? 6) 6.42 6.50 γ-Linoleicacid (C18: 3 Ω6) 0.09 0.13 Linolenicacid (C18: 3? 3) 28.75 27.86 Eicosenoic acid (C20: 1? 9) 0.28 0.23 Arachidonic acid (C20: 4? 6) 0.00 0.00 Eicosapentaenoic acid (EPA) (C20: 5? 3) 0.00 0.00 Docosatetraenoicacid (C22: 4? 6) 0.00 0.00 Docosahexaenoicacid (DHA) (C22: 6? 3) 0.00 0.00 Fatty acids (total) 100.00 100.00 Saturated fatty acid 32.51 33.21 Unsaturated fatty acid 67.49 66.79 - monovalent fatty acids 32.23 32.31 - polyvalent fatty acids 35.27 34.49

division Take a nap before learning Snooze Ca (calcium) 197.83 198.90 Cu (copper) 1.04 1.20 Fe (iron) 8.49 3.17 K (potassium) 1176.37 1045.44 Mg (magnesium) 211.94 199.25 Mn (manganese) 1.70 1.60 Na (sodium) 11.03 14.93 P (in) 749.79 688.27 Zn (zinc) 5.99 5.76 S (sulfur) 289.61 327.00

From the above results, it can be seen that the content of various constituents in the awakening before being cooked has been variously changed as the contents of the various ingredients have become familiar. In particular, the content of crude protein in Table 1 and the content of serine, glycine and alanine in Table 2 are remarkably increased, and it can be confirmed that the pharmacological effect of the above-described snooping will be changed due to the change in the composition of the cooked snack .

≪ Example 3: Preparation of experimental animals &

Seven-week-old SD rats were used for the non-alcohol-induced hepatitis test. Six to ten rats per group were prepared. For the SD rats, rodent diet AIN-76A was used. To this AIN-76A, a silkworm powder having the silk protein of Example 1 was mixed with 0.1 to 10 g / kg of SD rat per day. Diethylnitrosamine (DEN) (100 mg / kg) was administered 14 days later, and the change in body weight was checked again after 18 hours. Silkworm powder with silkworm protein was prepared from three white-jade, golden-silk and light-green varieties.

≪ Example 4: Confirmation of liver tissue status >

14 days was silkworm powder is treated with a Diethylnitrosamine the administration SD rat 100 mg per kg body weight of SD rat with a silk proteins, after 18 hours were collected the blood liver to CO ₂ euthanasia and necropsy, observed gross finding among the groups Respectively.

1, it was confirmed that weight and liver weight were slightly decreased after administration of diethylnitrosamine (DEN) (right side of FIG. 1A, FIG. 1B, FIG. 1C), and hepatocyte necrosis, inflammation, However, it was observed that these pathological symptoms were reduced in the silkworm powder administration group with silk protein (left side in FIG. 1B), and the white silk had the best liver protective effect in particular.

Example 5. Analysis of pathological score by H & E staining.

In each group of euthanized SD rats in Example 4, liver tissues were extracted and fixed in 10% formalin, fixed tissues were cut to a certain thickness, and paraffin embedded tissues were prepared. Hematoxylin and Eosin Histopathological findings of fatty liver were observed by blind test. The results are shown in FIG. 2.

Hematoxylin & Eosin staining is done by using hematoxylin and eosin, which are dark blue and red. The nuclei are stained in blue and most cytoplasmic components are stained pink or red. By using this staining method, the nucleus and cytoplasm of a cell can be observed. Referring to the result of Hematoxylin & Eosin staining shown in FIG. 2, it was confirmed that the group administered with silkworm powder containing diethylnitrosamine (DEN) and the silk protein recovered to a liver tissue state similar to the control (Normal) state.

Example 6: Identification of blood liver damage factor

Blood was collected from each group of euthanized SD rats in Example 4 and ALP (Alkaline Phosphatase), ALT (ALT) or Serum Glutamate Pyruvate Transaminase (AST) and ASST (AminoTransferase) or Serum Glutamic Oxaloacetic Transaminase And these are shown in Figs. 3A to 3C. For this purpose, serum was separated from the blood, and liver damage markers such as ALP, ALT and AST were measured using an Olympus Diagnostics AU480 Chemistry Analyzer.

3, the levels of ALP, ALT and AST were elevated in the blood of diethylnitrosamine (DEN) administration group. However, in the group pretreated with the silk protein-containing silk protein of Example 1, ALP, ALT, and AST levels were statistically significantly reduced, indicating that the silkworm powder having silk protein of Example 1 has excellent liver protecting effect.

≪ Example 6: Confirmation of inflammation-inhibiting effect &

Example 6-1. COX-2 and iNOS expression levels

Protein levels of COX-2 and iNOS in the liver of SD rats euthanized in Example 4 were measured by Western blot and the results are shown in FIG.

For this purpose, liver tissue obtained after autopsy was transferred to a mortar in the presence of liquid nitrogen, and cell lysis buffer and proteinase inhibitor were added to separate the proteins. Separated proteins were separated by protein size using SDS gel after quantitative determination of protein samples per group. The proteins in the gel were transferred to a PVDF membrane and reacted with an antibody of the desired protein to confirm presence or expression of the protein. Each experiment was statistically performed by performing at least 3 samples per group.

4, the expression of COX-2 and iNOS was increased in SD rats alone treated with diethylnitrosamine (DEN). However, in the SD rats pretreated with the silkworm protein silk protein of Example 1, their expression was remarkable And the production of inflammatory substances was suppressed.

Example 6-2. F4 / 80 and CD31 expression levels

In order to examine the expression level of the cells mediating the inflammatory reaction, macrophages were stained with F4 / 80 antibody in the liver tissues of euthanized experimental animals in Example 4, T cells were stained using CD31 antibody, Is shown in Fig. After the autopsy, liver tissue was removed and fixed in 10% formalin, fixed tissue was cut to a certain thickness, and paraffin embedding was performed to prepare tissue section slides. Each slide was immersed in xylene to remove paraffin, hydrated, and then subjected to antigen retrieval using an appropriate buffer. Next, the primary antibody of the protein to be expressed may be reacted, the secondary antibody capable of binding to the primary antibody may be reacted, and the protein may be stained after treatment with the coloring reagent binding to the secondary antibody. Each of the dyed samples was observed by a skilled experimenter using a microscope and a score for each staining was prepared. 0, 1, 10, 2, 10, 50, and 50%, respectively, for 0%, 10%, and 10% of the stained inflammatory mediators.

5, significant increases in macrophages and T cells were observed in the diethylnitrosamine (DEN) administration group, but it was confirmed that the administration amount of silkworm silkworm protein significantly decreased the expression level thereof.

Example 6-3. Identifying T cell activation factors

MRNA levels of the T cell activation-related factors were measured and the results are shown in FIG. ** T cell activation factors: interleukin-1beta (IL-1β), interferon gamma (IFN-γ), CXC motif chemokine 11 (CXCL11), chemokine (CC motif) ligand 17 (CCL17), CC chemokine receptor type 2 or CD192).

For this purpose, liver tissue obtained after autopsy was transferred to a pellet bowl in the presence of liquid nitrogen, and RNA was isolated using Trizol and isopropanol. The isolated RNA was synthesized as cDNA after confirmation of purity and the degree of expression of various genes was confirmed by PCR. Each experiment was statistically performed by performing at least 3 samples per group.

6, significant increases in T cell activation factors were observed in the diethylnitrosamine (DEN) administration group, but it was confirmed that the administration amount of silkworm silkworm protein significantly decreased the expression level thereof . The effect of diethylnitrosamine (DEN) on various factors that activate T cell to induce inflammation reaction and hepatotoxicity in the liver is suppressed by pretreatment of silkworm silkworm powder with immunity I can do it.

≪ Example 7: Comparison of effects of various silkworm-related powders &

The silkworm powder having the silk protein of Example 1, the white silk powder (the dried silkworm died from dead powdery mildew) (purchased from the Chinese medicine hospital), the silkworm silkworm cow silkworm powder obtained by the method of Example 1, The pupa was pulverized by the method of Example 1, and the silk protein powder in the circulation was fed for 14 days. Diethylnitrosamine (DEN) was administered by the method of Example 3 and euthanized 18 hours later. TNF-α expression was measured (using the kit provided by R & D systems).

Referring to the results shown in Table 5, it is confirmed that the TNF-a reduction effect of the powder of Example 1 of the present invention is the most superior to that of other powders.

Condition TNF-a (pg / ml) Untreated group 6.4 Diethylnitrosamine (DEN) alone group 16.2 Diethylnitrosamine (DEN) administration after powder administration of Example 1 8.7 Diethylnitrosamine (DEN) administration after saccharide powder administration 15.3 Diethylnitrosamine (DEN) administration after 5th day of silkworm powder administration 13.2 Diethylnitrosamine (DEN) treatment after pupa powder 16.0 Diethylnitrosamine (DEN) after silk protein powder administration 14.1

≪ Formulation Example 1 >

200 g of silkworm silkworm silkworm protein of Example 1 (cooked silkworm powder) was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. To this mixture was added a 10% gelatin solution, which was pulverized and passed through a 14-mesh sieve. This was dried, and a mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into tablets.

<Formulation Example 2: Food Preparation>

Formulation Example 2-1. Manufacture of cooking seasonings

A silkworm having the silk protein of Example 1 (a safflower powder) was added to the cooking sauce at 1% by weight to prepare a cooking sauce for health promotion.

Formulation Example 2-2. Manufacture of flour food products

A food for health promotion was prepared by adding breadcrumbs, cakes, cookies, crackers and noodles using the mixture of silkworm silkworm silkworm protein of Example 1 (wheat silkworm powder) added to flour at 0.1 wt%.

Preparation Example 2-3. Manufacture of soups and gravies

Health enhancing soup and juice were prepared by adding silkworm silkworm protein of Example 1 (cooked silkworm powder) to soup and juice at 0.1 wt%.

Formulation Example 2-4. Manufacture of dairy products

Silkworms (silkworm powder) having the silk protein of Example 1 were added to milk in an amount of 0.1 wt%, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation Example 2-5. Vegetable juice manufacturing

0.5 g of silkworm silkworm protein silkworm (the silkworm powder obtained in Example 1) was added to 1,000 ml of tomato juice or carrot juice to prepare health promotion vegetable juice.

Formulation Example 2-6. Manufacture of fruit juice

Healthy fruit juice was prepared by adding 0.1 g of silkworm silk protein (silkworm powder) of Example 1 to 1,000 ml of apple juice or grape juice.

Claims (12)

Four silkworms from 4th day after 4th day of silkworm to silkworm silkworm selected from silkworm silkworms are incubated in water for 30 to 24 hours at 55 ~ 125 ℃, Or a pharmaceutically acceptable salt thereof, for the prophylaxis or treatment of nonalcoholic hepatitis. The method according to claim 1,
The composition for preventing or treating non-alcoholic hepatitis is characterized in that the hepatitis is acute hepatitis or chronic hepatitis. The method according to claim 1,
A composition for preventing or treating nonalcoholic hepatitis characterized in that the silkworm having silk protein is a silkworm silkworm. delete The method according to claim 1,
The method for preventing or treating non-alcoholic hepatitis according to any one of claims 1 to 3, further comprising drying the silkworm having the silk protein protein, followed by drying through hot air drying or vacuum freeze drying. The method according to claim 1,
Wherein the picked silkworm is extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof to prepare an extract as a non-alcoholic hepatitis. Four silkworms from 4th day after 4th day of silkworm to silkworm silkworm selected from silkworm silkworms are incubated in water for 30 to 24 hours at 55 ~ 125 ℃, Or a pharmaceutically acceptable salt thereof, for the prevention or amelioration of nonalcoholic hepatitis. 8. The method of claim 7,
Wherein the hepatitis is acute hepatitis or chronic hepatitis. The health functional food for preventing or improving non-alcoholic hepatitis. 8. The method of claim 7,
Wherein the live silkworm having the silk protein is a silkworm of the silkworm. delete 8. The method of claim 7,
The method of any one of claims 1 to 3, further comprising a step of drying the raw silkworm having the silkworm protein, followed by drying through hot air drying or vacuum freeze drying, thereby pulverizing the raw silkworm. 8. The method of claim 7,
Characterized in that the picked silkworm is extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof to prepare an extract form.

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