KR20100074738A - Herbal extract for prevention or treatment of inflammatory diseases and pharmaceutical composition and functional food containing the same - Google Patents

Abstract

PURPOSE: A composition containing herb extract is provided to prevent or treat various inflammatory diseases caused by IL-32. CONSTITUTION: A herb extract for preventing or treating inflammatory diseases contains extract of discoreae Rhizoma, liriopis Tuber, asparagus cochinchinensis, polygalae Radix, acori graminei Rhizoma, zizyphi spinosi Semen, longanae Arillus, scutellariae Radix, radish seed, thujae Semen and Chrysanthemi Flos. The herb extract contains quercetin. The herb extract suppresses IL-32 expression. A pharmaceutical composition for preventing or treating inflammatory diseases contains the herb extract as an active ingredient.

Description

Herbal extracts, pharmaceutical compositions and functional foods for the prevention or treatment of inflammatory diseases {HERBAL EXTRACT FOR PREVENTION OR TREATMENT OF INFLAMMATORY DISEASES AND PHARMACEUTICAL COMPOSITION AND FUNCTIONAL FOOD CONTAINING THE SAME}

The present invention relates to novel herbal extracts and pharmaceutical compositions and functional foods containing the same, which exhibit excellent prophylactic or therapeutic efficacy against various inflammatory diseases caused by the inflammatory cell activator IL-32.

Inflammatory responses are caused by organized reactions of inflammatory cells through complex chemical signals and are increased by reactions such as trauma, infection, hypoxia, toxicity, allergies and autoimmune damage. In chronic inflammatory diseases, inflammatory reactions cause and sustain tissue damage, and infiltration of monocytes secreting inflammatory mediators, including inflammatory cell activators, has been reported at the site of inflammatory reactions.

Expression of inflammatory cell activators is regulated by the Nuclear Factor-kB (NF-kB) activity, a transcription factor. NF-kB binds to a consensus sequence present at the promoter position of a target gene (mainly an inflammatory cell activator) and initiates transcription of the inflammatory cell activators TNF-α and IL-6. NF-kB generally binds to the cytoplasm with IkB, an inhibitory protein, and when activated, breaks down the IkB protein and transfers it into the nucleus. The migrated NF-kB binds to the DNA of the target gene and increases transcription of the target gene. Recently, cell experiments in which caspase-1 deficient macrophages and caspase-1 were inactivated resulted in the suppression of NF-kB activity by LPS. I found it via -1.

Caspase-1 is a type of cystein-aspartic acid protease. Caspase-1 is an enzyme that activates interleukin-1beta and interleukin-18, a cytoactive substance involved in the inflammatory response. Caspase-1 has an N-terminal caspase recruitment domain (CARD), which enhances caspase activity in apoptosis and inflammation. Receptor interacting protein-2 (RIP2) modulates caspase-1 activity through CARD-CARD binding. RIP2 induces NF-kB activity by binding to IkB kinase (IKK) -γ to form an IKK conjugate. Recently, caspase-1 has been reported to increase the expression of IL-32, an inflammatory cell activator.

IL-32, also known as NK cell transcrpt 4 (NK4), is a natural killer that is activated by activated lymphocytes, epithelial cells activated by interferon-gamma (IFN-γ), IL-12, IL-18 and IL-32. Produced by the cell. Human IL-32 includes IL-32α, IL-32β, IL-32γ, IL-32δ, IL-32ε and IL-32ζ. Recombinant IL-32 induces the production of IL-1β, IL-6, IL-8 and TNF-α by the activity of NF-kB and mitogen-activated protein kinase (p38 MAPK). It has been reported that the maturation of IL-1β through caspase-1 dependent mechanisms is also mediated by IL-32, and the production of IL-32 is inflammation including IL-18 activated through caspase-1 dependent mechanisms. It is reported to be caused by mechanism. IL-32 is expressed in the synovial membrane of patients with rheumatoid arthritis and is known as a factor that attenuates the disease along with other inflammatory cell activators. IL-32α expression is increased in mucosal membranes of enteritis and Crohn's patients. It has been reported to augment arthritis or colitis through feedback action with -α. Recently, blocking the expression of tumor necrosis factor receptors in rats suffering from colitis inhibited the synthesis of TNF-α by IL-32, improving the inflammatory response. These results indicate that IL-32 plays an important role in the inflammatory response.

Accordingly, the present inventors inhibited the expression of IL-32 in peripheral blood mononuclear cells (PBMCs), inhibits caspase activity and blocks the migration of NF-kB into the nucleus, thereby inhibiting the inflammatory response by lipopolysaccharide (LPS). To develop a herbal extract that can inhibit the production of inflammatory cell activators IL-8 and TNF-α produced by IL-32 stimulation in THP-1 cells, and the inhibitory effect of IL-32 production The present invention has been completed by pharmacologically observing the inhibitory mechanism, confirming that the extract has an anti-inflammatory effect.

The present invention is to provide a novel herbal extract and a pharmaceutical composition and health functional food comprising the same, which shows excellent efficacy against various inflammatory diseases caused by the inflammatory cell activator IL-32.

The present invention for achieving the above object is an extract of a mixture containing lotus root, medicinal herb, macmundong, astronomical dong, Wonji, Seokchangpo, Sanjoin, Yongan, golden, nachojakja, white porcelain and gamguk, herbal extract for preventing or treating inflammatory diseases To provide.

In addition, the mixture is lotus root, acid, munmundong, astronomical dong, Wonji, Seokchangpo, Sanjoin, longan meat, golden, Nabokjaja, white porcelain and gamguk 1-10: 1-10: 1-7: 1-7: 1-7: 1 -7: 1-7: 1-7: 1-7: 1-7: 1-7: 1-3 to provide an herbal extract for preventing or treating inflammatory diseases, characterized in that mixed.

In addition, the mixture is lotus root, powder, makmundong, astronomical dong, Wonji, Seokchangpo, Sanjoin, longan meat, gold, nabokjaja, white porcelain and gamguk 6: 6: 3: 3: 3: 3: 3: 3: 3: 3 Provided is an herbal extract for preventing or treating inflammatory diseases, which is mixed at a weight ratio of 1: 1.

The present invention also provides a herbal extract for preventing or treating inflammatory diseases, which contains quercetin.

In addition, the present invention provides an herbal extract for preventing or treating inflammatory diseases, which is characterized by suppressing the expression of Interleukin-32 (IL-32).

The present invention also provides a pharmaceutical composition for preventing or treating inflammatory diseases, containing the herbal extract as an active ingredient.

The present invention also provides a dietary supplement for preventing and treating inflammatory diseases, which contains the herbal extract and includes a food supplement which is food acceptable.

The herbal extracts, pharmaceutical compositions, and dietary supplements according to the present invention inhibit IL-32 production and IL-32γ mRNA expression by LPS, inhibit inflammatory cell activator production by LPS, and increase by LPS. Reduced mRNA expression levels. It also effectively inhibits inflammatory responses by inhibiting NF-kB / DNA binding capacity, inhibiting the degradation of caspase-1 precursors, and inhibiting IL-8 and TNF-α production increased by IL-32 stimulation. can do.

Hereinafter, with reference to the accompanying drawings, an embodiment of the present invention for preventing or treating inflammatory diseases, herbal extracts, pharmaceutical compositions, and dietary supplements will be described in more detail.

First, the herbal extract according to the present invention will be described.

The herbal extracts of the present invention are water extracts or solvent extracts or fractions thereof of a mixture of lotus root, medicinal herb, macmundong, astronomical dong, Wonji, Seokchangpo, Sanjoin, longan meat, golden, nabokjak, white porcelain, and persimmon soup.

One of the mixture of the lotus root is known to have a pharmacological action, such as anti-thrombosis, anti-inflammatory, sedative action, and the acid medicine is known to have a hypoglycemic, immune boosting, intestinal motility, antioxidant activity.

In addition, the pulmonary dong is known to have a continuous hypoglycemic, anti-inflammatory action, astronomical dong is known to have antibacterial, anti-cancer, immune enhancing action.

In addition, the base is known to have a calming and anti-empty, expectorant, uterine contraction, lowering blood pressure, antibacterial effect, Seokchangpo inhibits the central nervous system, muscle relaxation and spasms, promotes the secretion of digestive fluids, inhibits abnormal gastric fermentation, antibacterial It is also known to reduce blood lipids.

Sanjoin is also known for sedation and hypnosis, antipain, analgesic and hypothermia, antiarrhythmia, myocardial ischemia, blood pressure and hemoglobin, immune function, antiplatelet aggregation. It is known to have an immune enhancing, antibacterial, anticancer and anti-aging effect.

In addition, the gold and silver antiviral and antiviral, anti-inflammatory and anti-hypersensitivity, sedation and blood pressure lowering, lipid lowering, interpolation, edema, antioxidant, anticoagulant and antithrombotic, promoting cell immunity and anti-tumor, diuresis, myrrh, cataract prevention action It is known that nabokjak has the effect of improving intestinal motility, Jinhae, expectoration, Pyeongcheon, antibacterial, anti-inflammatory, lowering blood pressure.

In addition, the white porcelain is known to have a memory improvement, sedation, and the country is known to have increased blood flow, lowering lipid lipids, antibacterial, central nervous system.

The Yeonjakgyeon, mountain medicine, makmundong, astronomical dong, Wonji, Seokchangpo, Sanjoin, longan meat, gold, Nabokjaja, white porcelain and gamguk 1-10: 1-10: 1-7: 1-7: 1-7: 1-7: 1 Mixing at a weight ratio of -7: 1-7: 1-7: 1-7: 1-7: 1-3 is preferred because excellent anti-inflammatory efficacy can be obtained. More preferably, it is mixed in a weight ratio of 6: 6: 3: 3: 3: 3: 3: 3: 3: 3: 3: 1. When mixed in the weight ratio, it is possible to obtain a better anti-inflammatory effect than mixed in other weight ratio.

The method of obtaining the extract according to the present invention by extracting the mixture as described above is not particularly limited, and may be extracted according to various methods using water or an organic solvent. For example, the herbal extract of the present invention can be obtained by extracting the herbal mixture mixed in the above weight ratio with water or an organic solvent and fractionating with ethyl acetate.

That is, the herbal extract may be prepared by immersing the pulverized product for each extraction region in distilled water or an organic solvent, filtering and drying the solvent. Alcohol, hexane, chloroform and the like may be used as the organic solvent. The alcohol solvent is preferably an alcohol having 1 to 5 carbon atoms, more preferably ethanol, methanol, propanol or butanol, and most preferably ethanol and methanol. The extraction part may be a flower, a leaf, a stem, a bark, a bud, a root, a fruit, a seed or a plant of the plant, and preferably, lotus root, sanjoin, or moth is a seed, and the medicine, macmundong, astronomical dong, Wonji, Seokchangpo and golden It is the root, Yongan flesh is abyssal, white porcelain is species, and gamguk is flower.

In addition, the extract is first, pulverized plants or dried products of plants, heat extraction and concentration by adding a solvent in a weight ratio of 1: 1 to 1:25 to the pulverized product, and then filtered or centrifuged to obtain a supernatant You can also get The heat extraction temperature is preferably 90 ~ 110 ℃, it is preferably made for 0.5 to 10 hours.

The supernatant can be fractionated with ethyl acetate (layer separation) to obtain a herbal extract. It is preferable to perform the said fraction 3 times or more.

The herbal extract may be used in liquid form, or may be used as a powder in which the solvent is removed by vacuum concentration or lyophilization.

According to the present invention, the herbal extract of the mixture of lotus root, medicinal herb, macmundong, astronomical dong, Wonji, Seokchangpo, sanjoin, longan meat, golden, nabokjak, white porcelain and gamguk is lipopolysaccharide (LPS; lipopolysaccharide) And inflammation caused by inflammatory cell activators. The herbal extract contains a quercetin (Quercetin) known as an anti-inflammatory substance as a component.

More specifically, the herbal extract according to the present invention inhibits IL-32 production and IL-32γ mRNA expression by LPS, inhibits inflammatory cell activator production by LPS, and increases the mRNA expression level increased by LPS. Can be reduced. In addition, by inhibiting NF-kB / DNA binding capacity, inhibiting the degradation of caspase-1 precursor, and inhibiting the production of IL-8 and TNF-α increased by IL-32 stimulation, it is possible to effectively suppress the inflammatory response. have.

The present invention also provides a pharmaceutical composition for the prevention and treatment of inflammatory diseases comprising the herbal extract as an active ingredient. The pharmaceutical composition may further include pharmaceutically acceptable ingredients that do not inhibit the anti-inflammatory activity of the herbal extract.

Suitable formulations of the pharmaceutical compositions include, but are not limited to, powders, solutions, pills, tablets, dragees, hard or soft capsules, solutions, suspensions or emulsions, injections, suppositories, and the like. The pharmaceutical composition may be prepared in a suitable formulation using a pharmaceutically inert organic or inorganic carrier with a pharmaceutically acceptable salt. That is, when the formulation is a tablet, coated tablets, dragees and hard capsules, lactose, corn starch or derivatives thereof, talc, stearic acid or salts thereof can be used. Also, when the formulation is a soft capsule, polyols of vegetable oils, waxes, fats, semisolids and liquids can be used. In the case of solution or syrup form, water, polyols, glycerol and vegetable oils and the like can be used. As suppository carriers, natural or hardened oils, waxes, fats, liquid polyols and the like can be used.

The present invention also provides cosmetics, pharmaceuticals and the like processed with the pharmaceutical composition of the present invention. The pharmaceutical product of the present invention is not particularly limited in its formulation, and may be formulated into conventional oral, parenteral, and topical administration according to the purpose. In the preparation for oral administration of powders, granules, tablets, capsules and the like, binders, suspending agents, disintegrating agents, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, flavoring agents and the like can be used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents, water and liquid paraffin. In the case of injections, buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers and the like can be used in combination. In addition, it may be formulated into an ointment, a spray, a lotion, a patch, a pad, a cream.

The present invention also provides a dietary supplement for the prevention and improvement of inflammatory diseases, including the herbal extract and a food supplement acceptable food supplement. The herbal extract according to the present invention can be used as a food grade, so that it can be safely ingested without a separate purification process for removing toxicity, so it has an anti-inflammatory activity, lotus root, medicinal herb, mackmundong, astronomical dong, Wonji, Seokchangpo, Sanjoin, Yongan Ingestion of the health functional food of the present invention, including the herbal extract of golden, moth, white porcelain, gamgu mixture, can obtain anti-inflammatory effect.

The health functional food may contain the herbal extract in the form of a pulverized product, liquid extract or powder extract, and may be used in the form of beverages, gums, teas, vitamin complexes, health supplements, tablets, capsules, etc. It may also be prepared in the form of powders, granules, liquids, pills and the like.

The present invention is described in more detail based on the following examples or experimental examples, but the present invention is not limited thereto.

Example 1 Preparation of Herbal Extracts 1

Prepare 60g each of dried lotus root and mountain roots, 30g each of Macmundong root, Cheonmundong root, native root, Seokchangpo root, Sanjoin seed, Longan meat seed skin, golden root, moth seed and white porridge. Mixed.

400 g of the mixture was poured into 1 L of water, and the mixture was shaken for 3 hours, and then reheated under the same conditions. The concentrated solution was filtered through a 0.45 μm filter and then lyophilized to obtain the result.

Example 2 Preparation of Herbal Extracts 2

Dried lotus root, powder, macmundong, astronomical dong, Wonji, Seokchangpo, sanjoin, longan meat, golden, nabokja, white porcelain, and Korean soup 6: 6: 3: 3: 3: 3: 3: 3: 3: 3: 1 After mixing at a weight ratio of 4 kg of the crude drug, it was repeated three times for 4 hours in 20% methanol to extract the reflux cooling hot water. After filtering the hot water extract, the solvent was removed by a vacuum condenser to obtain an extract 765g. After adding 10% methanol to the methanol extract and suspending it, the fractions were systematically fractionated using CH 2 Cl 2 (methylene chloride), EtOAc (ethyl acetate), n-BuOH (n-butanol), and H ₂ O (water). 34 g of CH ₂ Cl ₂ fraction, 34 g of EtOAc fraction, 53 g of n-BuOH fraction, and 352 g of H ₂ O fraction were obtained.

The ethyl acetate fraction (30 g) was chromatographed on silica gel (560 g) column (6.3 × 79 cm), and the mixture of CH ₂ Cl ₂ and CH ₂ Cl ₂ -methanol-water as an eluting solvent (5: 1: 1, 25: 8: 5, 7: 3: 1, 65,35,10, lower layer), eluting with increasing polarity and dividing into 10 subfractions according to the TLC pattern. The resultant was obtained from the small fraction.

Example 3 Preparation of Herbal Extract 3

Dried lotus root, powder, macmundong, astronomical dong, Wonji, Seokchangpo, sanjoin, longan meat, golden, nabokjak, white porcelain, and Korean soup 10: 10: 5: 5: 5: 5: 5: 5: 5: 5: 5: 1 The mixture was mixed in a weight ratio of 4 kg, and heating extraction was repeated three times with 20% methanol. The extract was concentrated under reduced pressure to obtain 1200 g of extract. The methanol extract was systematically fractionated with CHCl ₃ (trichloromethane), EtOAc (ethyl acetate), n-BuOH (n-butanol), n-Hexane (n-hexane).

120 g of the ethyl acetate fraction was subjected to silica gel column chromatography (hexane → hexane: ethyl acetate → ethyl acetate: methanol → methanol) to obtain seven small fractions (A1-7). A5 was subjected to silica gel column chromatography (trichloromethane: methanol: water = 25: 4: 1) to obtain 8 fractions (B1-8), and B8 was subjected to Sephadex LH-20 column chromatography (methanol). The resultant was repeated six times, followed by silica gel column chromatography (trichloromethane: methanol: water = 25: 4: 1).

Example 4 Preparation of Herbal Extracts 4

Dried lotus roots, potions, macmundong, astronomical dong, Wonji, Seokchangpo, sanjoin, longan meat, golden, nabokja, white porcelain, and gamguk 10: 10: 3: 3: 3: 3: 3: 3: 3: 3: 1 The mixture was mixed at a weight ratio of 4 g, and these 4 g of crude medicines were ground and powdered, and then extracted twice with 30 mL of water at 60 ° C. for 30 minutes, followed by filtration. The extract was concentrated under reduced pressure at 45 ° C. to obtain a brown extract. The brown extract was resuspended with 50 mL of water and partitioned twice with 50 mL of ethyl acetate. 5 mL of acetic acid was added to the H ₂ O-soluble fraction, 1-butanol was added to the acidic water fraction, and the fractions were divided. The 1-butanol fraction was concentrated under reduced pressure at 40 ° C. to obtain 1.0 g. This was recrystallized using water / 2-propanol / acetic acid (4/3/1) to give the result.

The resulting products (medicinal extracts) obtained according to Examples 1 to 4 were named HS2008, and HS2008 obtained according to Example 1 was used as a sample in the following Experimental Example. For reference, using the HS2008 obtained according to any one of Examples 1 to 4 shows similar results to the results according to the following experimental example.

Experimental Example 1. The effect of the expression of inflammatory cell activator increased by LPS

Experimental Example 1-1. IL-32 Production and Expression Regulation Effects

First, the cells to be used for the experiment were stabilized for 3 hours by dispensing 2 × 10 ⁵ cells / ml in a 24-well cell culture plate, and the stabilized cells were quercetin which is a component of HS2008 and HS2008 of the present invention for 2 hours. Were pretreated, and LPS was reacted for 24 hours before centrifugation. The supernatant obtained by centrifugation was quantified using an IL-32 antibody and the absorbance was measured by an ELISA analyzer at a wavelength of 405 nm.

1A is a graph showing the effect of HS2008 and quercetin on the production of IL-32 by LPS in PBMCs. As shown in Figure 1a, it can be seen that HS2008 and quercetin inhibit the inflammatory response by inhibiting IL-32 production by LPS.

Next, IL-32 mRNA expression levels were analyzed using the RT-PCR method. First, the cells to be used in the experiment was stabilized for 3 hours by dispensing 2 × 10 ⁶ cells / ㎖ in the cell culture plate, the stabilized cells were pretreated with HS2008 of the present invention for 2 hours and LPS for 8 hours. After total RNA was isolated from the cells to synthesize cDNA, polymerase chain reaction (PCR) on inflammatory cell activators was performed to analyze the mRNA expression level of IL-32 by HS2008.

Figure 1b is a photograph showing the effect of HS2008 and quercetin in the expression of IL-32γ mRNA by LPS, Figure 1c is a graph quantifying the data of Figure 1b using an image analyzer. As shown in FIG. 1B and FIG. 1C, it can be seen that HS2008 effectively inhibits IL-32γ mRNA expression by LPS.

Experimental Example 1-2. Inflammatory cell activator production regulation effect

First, the cells to be used in the experiment were stabilized for 3 hours by dispensing 2 × 10 ⁵ cells / ml in a 24-well cell culture plate, and the stabilized cells were subjected to HS2008 and constituent quercetin of the present invention for 2 hours by concentration. Pretreatment, LPS was reacted for 24 hours and centrifuged. The supernatant obtained by the centrifugation was quantitated inflammatory cytokines IL-1β, IL-6, IL-8 and TNF-α using an antibody and the absorbance was measured by ELISA analyzer at a wavelength of 405nm. The absorbance measured was expressed by inhibition rate.

Table 1 below shows data for quantifying inflammatory cell activators increased by LPS.

Table 1: Effect of HS2008 on the production of inflammatory cell activators by LPS stimulation

HS2008 (mg / ml) IL-1β (ng / ml) IL-6 (ng / ml) TNF-α (ng / ml) Blank 0.95 ± 0.17 2.13 ± 0.24 0.39 ± 0.04 LPS 1.37 ± 0.06 14.33 ± 4.55 0.74 ± 0.19 LPS + HS2008 (0.01) 1.30 ± 0.19 13.80 ± 1.3 0.61 ± 0.21 LPS + HS2008 (0.1) 1.15 ± 0.26 8.68 ± 2.8 0.57 ± 0.18 LPS + HS2008 (1) 0.60 ± 0.23 4.28 ± 0.86 0.49 ± 0.04

As shown in Table 1, it can be seen that the production of inflammatory cell activators by LPS stimulation in PBMCs is inhibited when HS2008 is added, especially when the HS2008 is added in an amount of 1 mg / ml.

2 is a graph showing the inhibitory effect of HS2008 on the production of inflammatory cell activators by LPS in PBMCs, Figure 4 is a graph showing the inhibitory effect of quercetin on the production of inflammatory cell activators by LPS in PBMCs. . 2 and 4 it can be seen that HS2008 inhibits the inflammatory response by inhibiting the production of inflammatory cell activators.

Experiment 1-3. Inflammatory cell activator expression control effect

Inflammatory cell activator mRNA expression levels were analyzed using the RT-PCR method. First, the cells to be used for the experiment was stabilized for 3 hours by dispensing 2 × 10 ⁶ cells / ㎖ in a cell culture plate, the pretreated HS2008 of the present invention to the stabilized cells for 2 hours and then treated with LPS for 8 hours. After cDNA was synthesized by separating total RNA from the cells, mRNA expression levels of inflammatory cell activators by HS2008 were analyzed by polymerase chain reaction (PCR).

Figure 3 is a photograph showing the effect of HS2008 on the mRNA expression of inflammatory cell activators by LPS in PBMCs. As shown in FIG. 3, HS2008 decreased the mRNA expression level increased by LPS.

Experimental Example 2. Effect of LPS on the mechanism of inflammatory mechanism

Experimental Example 2-1. NF-kB activity inhibitory effect

Since NF-kB activity is related to the inflammatory response, the effect of HS2008 on the NF-kB activity was observed. Western blot analysis data of Figure 5a and Figure 5c shows that HS2008 inhibits the migration of NF-kB into the nucleus, it can be seen that also inhibits the degradation of the NF-kB inhibitor IkBα. In addition, NF-kB / DNA binding capacity was measured using a transcription factor-enzyme linked assay (TF-EIA) method to confirm the effect of HS2008 on NF-kB activity. Oligonucleotide salts with NF-kB binding sites were used labeled with biotin. As a result, it was found that HS2008 also inhibited NF-kB / DNA binding ability (FIG. 5B).

Experimental Example 2-2. Caspase-1 Inhibitory Activity

Caspase-1 induces NF-kB activity and is activated by LPS. Therefore, the effects of HS2008 and quercetin on caspase-1 activity were examined. Caspase-1 quantitative kit was used to measure caspase-1 activity of extracts obtained from PBMCs. As a result, it was found that caspase-1 was activated by LPS. It can be seen that it is inhibited by quercetin (FIG. 6A). In addition, expression of caspase-1 precursor was examined using Western blot analysis. 6b shows that HS2008 and quercetin inhibit the degradation of caspase-1 precursor.

Experimental Example 3. Inflammatory cell activator expression regulation effect increased by IL-32 stimulation

First, the cells to be used in the experiment, human monocyte cell line THP-1 cells were dispensed by 2 × 10 ⁵ cells / ml in a 24-well cell culture plate and stabilized for 3 hours. The supernatant obtained by pretreatment with quercetin, bicalane and hyperside for 2 hours by concentration for 2 hours and reacting with IL-32 for 72 hours was used for quantification. Inflammatory cell activators IL-8 and TNF-α were quantified using the antibody, and the absorbance was measured by an ELISA analyzer at a wavelength of 405 nm. In Figure 7, it can be seen that HS2008, quercetin, bikallein and hypersides inhibit the inflammatory response by inhibiting increased IL-8 and TNF-α production by IL-32 stimulation.

From the above results, the herbal extract according to the present invention inhibits IL-32 production and IL-32γ mRNA expression by LPS, inhibits inflammatory cell activator production by LPS, and decreases the mRNA expression level increased by LPS. It can be seen that. It has also been shown to effectively inhibit inflammatory responses by inhibiting NF-kB / DNA binding capacity, inhibiting caspase-1 precursor degradation, and inhibiting IL-8 and TNF-α production increased by IL-32 stimulation. Can be.

Figure 1a is a graph showing the effect of HS2008 and quercetin on LPS-induced IL-32 production in PBMCs, Figure 1b is a photograph showing the effect of HS2008 and quercetin on IL32γ mRNA expression, Figure 1c Is a graph digitizing the photograph of FIG.

2 is a graph showing the inhibitory effect of HS2008 on the production of inflammatory cell activators by LPS in PBMCs.

3 is a diagram showing the effect of HS2008 on the mRNA expression of inflammatory cell activators by LPS in PBMCs.

4 is a graph showing the inhibitory effect of quercetin on the production of inflammatory cell activators by LPS in PBMCs.

Figure 5a is a diagram showing the effect of HS2008 on the nuclear and cytoplasmic NF-kB expression of PBMCs using Western blot analysis method, Figure 5b is HS2008 for DNA binding to NF-kB using TF-EIA method 5C is a graph showing the degree of cytoplasmic degradation of IkBα, an inhibitor of NF-kB, using Western blot analysis.

Figure 6a is a graph showing the quantification of caspase-1 activity by LPS in PBMCs, Figure 6b is a diagram showing the caspase-1 activity using Western blot analysis method.

Figure 7 shows the quercetin, bikallein and high components of HS2008 and HS2008 constituent herbal medicines for the production of inflammatory cell activators IL-8 and TNF-α by IL-32 stimulation in THP-1 cells, human monocytes It is a graph analyzing the effect of ferroside.

Claims (7)

An extract of a mixture containing lotus root, medicinal herb, macmundong, astronomical dong, Wonji, Seokchangpo, sanjoin, longan meat, gold, nabokja, white porcelain, and gamguk, herbal extract for preventing or treating inflammatory diseases. The method of claim 1, wherein the mixture is lotus root, medicinal herb, macmundong, astronomical dong, Wonji, Seokchangpo, Sanjoin, longan meat, golden, nabokjak, white porcelain and gamguk 1-10: 1-10: 1-7: 1-7: 1 -7: 1-7: 1-7: 1-7: 1-7: 1-7: 1-7: 1-3 The herb extract for preventing or treating inflammatory diseases, characterized in that mixed by weight ratio. The method of claim 1, wherein the mixture is lotus root, medicinal herbs, macmundong, astronomical dong, Wonji, Seokchangpo, Sanjoin, longan meat, golden, nabokjaja, white porcelain and gamguk 6: 6: 3: 3: 3: 3: 3: 3: 3 Herbal medicine extract for preventing or treating inflammatory diseases, characterized in that mixed in a weight ratio of 3: 3: 3. According to claim 1, Quercetin (Quercetin) containing a herbal extract for the prevention or treatment of inflammatory diseases. The herbal extract for preventing or treating inflammatory diseases according to claim 1, wherein the extract inhibits the expression of Interleukin-32 (IL-32). A pharmaceutical composition for preventing or treating inflammatory diseases, comprising the herbal extract of any one of claims 1 to 5 as an active ingredient. A health functional food for preventing or treating an inflammatory disease, comprising the herbal extract of any one of claims 1 to 5 and comprising a food supplement acceptable food supplement.

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