KR20100018174A - Skin external composition containing taraxacum platycarpum h. dahlsi, cimicifuga heracleifolia, dioscorea opposita or angelica tenuissima nakai - Google Patents

Abstract

PURPOSE: A skin external composition containing medicinal herb extract is provided to promote regeneration of wound tissue and suppress skin injury and aging. CONSTITUTION: A skin external composition for anti-inflammation contains medicinal herbs including Taraxacum platycarpum H. Dahlsi, Cimicifuga heracleifolia, Dioscorea opposite, or Angelica tenuissima Nakai extract as an active ingredient. The skin external composition for wound healing and anti-aging contains the medicinal herbs. The medicinal herb extract is obtained using water or organic solvent. The organic solvent is ethanol, methanol, butanol, ether, ethylacetate, or chloroform.

Description

Skin external composition containing Tarexacum platycarpum H. Dahlsi, Cimicifuga heracleifolia, Dioscorea opposita or Angelica tenuissima Nakai}

The present invention has an anti-inflammatory effect of inhibiting inflammation-related factors by containing one or more herbal extracts of pogongyoung, horse riding, powder and herbal extracts as an active ingredient, and promotes re-epithelialization of wound tissues to promote progression from inflammatory phase to proliferation. The present invention relates to an external composition for skin having an anti-aging effect of rapidly healing wounds and increasing gene expression of enzymes involved in detoxification, thereby inhibiting skin damage caused by external stimuli such as UV and inhibiting skin aging.

The skin acts as a shield to protect the body from the outside. When a wound occurs on the skin, the blood fills the wound site by the natural healing action of the living body, the granule reduction of platelets and activation of the hageman factor are initiated, and the wound healing process proceeds. Blood coagulation is a temporary protective action that protects exposed wound tissues and provides a basis for cells to migrate during the healing process (Lee SH AS, Jung SG. Skin Barrier. Seoul: Ryo Moon Gak, 2004).

The wound healing process is divided into four stages: inflammatory, re-epithelial, proliferative and mature stages. During the inflammatory phase, immune cells appear in the wound site, which have migrated from blood vessels to the wound site. Subsequently, growth factors and signaling substances that induce granulation formation are secreted. In the absence of serious infections, the inflammatory phase usually progresses briefly (Care KRGf. Advances in wound Care. Seoul: Korea Medical Book Publisher, 2002). Inflammation is an essential step in the healing process.

The proliferative phase occurs similarly to re-epithelialization, which is characterized by the formation of granular tissues at the wound site (Kubo KKY.Spongy matrix of hyaluronic acid and collagen as a cultured dermal substitute: evaluation in an animal test. J Artif Organs 2003; 6 (1) ): 64-70). Granulation tissue consists of a combination of extracellular matrix components such as immaturity collagen, fibronectin and hyaluronic acid, together with fibroblasts and inflammatory cells. It is important to treat the wound quickly and to have a structured structure. The exfoliated wound surface is covered by a layer of keratinocytes, creating a new epidermis and reconstructing the epidermal layer. Cells rise through the wound at the edge of the wound or the dermal residue of the skin left and begin migration through the crust and above the living connective tissue.

When the re-epithelialization of the wound is completed, a series of processes are performed in which the wound area is reduced through the increase and reorganization of connective tissue. Then, during the maturation phase, the convoluted cells and capillaries of the convalescent tissue gradually disappear, causing scarring if these tissues are hyperplastic or do not degrade normally. This is a common wound healing process. In the wound healing process, it is important not only to treat the wound quickly but also to treat it without side effects or scars, so that the tissue cells are balanced and balanced by suppressing inflammation and regulating the expression of growth factors. Efforts to find this continue.

Reduction in inflammation can be measured by decreasing the density of inflammatory cell types such as mononuclear cells or astrocytes, neutrophils, mast cells, basophil cells. Inflammation reduction can also be measured by measuring neutrophil activity (Jones et al., 1994), and factors such as the frequency of mast cell degranulation or the measurement of histamine levels or the level of reactive oxygen species are also measured as a measure of inflammation reduction. Can be used. The level of inflammation can also be determined by PCR for certain genes such as interferon-α, -β and -γ, tumor necrosis factor-α, interleukin 1β, -2, -4, -5, -6, -8, -12,- Can be measured indirectly by checking the transcriptional levels of genes such as 18, -23, -27, CD4, CD28, CD80, CD86, MHCII, and iNOS. Determination of proinflammatory cytokines levels in tissues and / or patients' fluids, such as plasma, may be a measure of inflammation reduction.

Recently, the correlation between the intrinsic inflammatory mediators of skin and aging has been reported. In particular, inflammatory mediators, tumor necrosis factor-α and interleukin 1β, are matrix metalloproteinases (MMPs) including collagenase-1 (MMP-1) and collagenase-3 (MMP-3). It has been reported to increase the production of and to decrease collagen and fibronectin synthesis (Wan et al., Transmodulation of epidermal growth factor receptor mediates IL-1 beta-induced MMP-1 expression in cultured human keratinocytes.Int J Mol 2001 Mar; 7 (3): 329-34; Tanaka et al., Prevention of the ultraviolet B-mediated skin photoaging by a nuclear factor kappaB inhibitor, parthenolide.J Pharmacol Exp Ther. 2005 Nov; 315 (2): Epub 2005 Jul 18; and Kong et al., Cyclophilin C-associated protein is a mediator for fibronectin fragment-induced matrix metalloproteinase-13 expression.J Biol Chem. 2004 Dec 31; 279 (53): 55334-40 Epub 2004 Oct 26).

Tumor necrosis factor-α and interleukin 1β have been reported to promote the degradation of intercellular matrix by inducing the production of MMPs in cells through the regulation of NF-κB, a transcription factor that regulates the synthesis of inflammatory signaling factors. Therefore, it may be suggested that inhibition of MMPs production and matrix protein production by tumor necrosis factor-α, interleukin 1β production and NF-κB activity is associated with collagen deficiency in aged skin. Drugs that inhibit the production of MMP-1 through reduced synthesis of inflammatory cytokines and reduced transcription factor NF-κB activity can be used as anti-aging agents.

Pogong-young (蒲公英, Taraxacum platycarpum H. Dahlsi) has been reported, such as dandelion, sulfide specified, or specified as referred pogong seconds or the like, makes a Reading function sakhineun the boil, anti-inflammatory, between protective efficacy and cadmium detoxification. In addition, jaundice, swelling, measles healing effect and the effect of blackening the hair is said to be. Korean Laid-Open Patent Publication No. 2002-44266 discloses a cosmetic composition comprising a complex herbal extract including pogongyoung, preventing skin aging and wrinkles, and having excellent whitening effect.

Horseback riding (升麻, Cimicifuga heracleifolia) has a function to rise upwards, it is called 'seung (rise) horse.' In the main river, horseback riding is hard. It clears the stomach, loosens the poison and goes up to make the pain of teeth go away. ' Its light and floating nature has the effect of raising the amount of yang up, the effect of releasing the table outside the body of the body and the effect of lowering and detoxifying heat. It is mainly used for headaches and toothaches caused by wind fever, and when the mouth is sore, when the throat hurts, it is used for the initial margin, marginal discharge of the mucous membranes or anterior layers of the anus or rectum. In addition, the strong detoxification action, such as gypsum and large blue leaves to suppress symptoms such as toothache or luminescence caused by stomach fever.

Sanos (山藥, Dioscorea opposita) has been widely used in medicine for a long time, such as dizziness, headache, sedation, stamina reinforcement, and phlegm removal, which has more than 10 known effects. It has a special effect on nourishing tonic and is effective in indigestion, gastrointestinal disorders, diabetes, cough and lung diseases. In particular, the kidney function is strong, strong people with weakened energy is good to take longer. They agree that they strengthen the muscles, stabilize the mind and strengthen the will.

Ancient book (Angel 本, Angelica tenuissima Nakai) relieves headaches caused by wind and avoids fog and dew poison. It is written in the agreement that it can revive flesh, improve face color, remove freckles, visa and acne, and make bath medicine and oil on face. In addition, Korean Laid-Open Patent Publication No. 2006-93164 discloses an anti-wrinkle cosmetic composition having excellent anti-wrinkle effect by promoting cell proliferation and collagen synthesis by including extracts of medicinal herbs of mung bean, white paper, white powder, cheonhwa powder, tomb and flake as active ingredients. It is.

Thus, the present inventors have measured wound healing in a cell culture state ( in By performing in vitro wound healing assay, pogongyoung, horseback riding, powder or herbal extract shows anti-inflammatory effect by inhibiting inflammation-related factors, and accelerates the progression from inflammatory phase to proliferation by promoting division and migration of keratinocytes. The present invention has been shown to have an effect of inhibiting skin aging by inhibiting skin damage caused by external stimuli such as ultraviolet rays by increasing gene expression of enzymes involved in detoxification with anti-inflammatory effects, and completing the present invention. Was done.

Accordingly, an object of the present invention has anti-inflammatory effects by inhibiting inflammation-related factors, and by promoting the division and migration of the keratinocytes of the skin to facilitate the progression from the inflammatory phase to the proliferative phase has a wound healing effect, external stimulation such as ultraviolet rays It is to provide a skin external preparation composition having the effect of inhibiting skin damage by inhibiting skin aging.

In order to achieve the above object, the present invention provides an anti-inflammatory composition containing as an active ingredient one or more herbal extracts selected from the group consisting of pogongyoung, horse riding, powdered and herbal extracts.

In another aspect, the present invention provides a wound healing composition containing as an active ingredient one or more herbal extracts selected from the group consisting of pogongyoung, horse riding, mountain powder and gobon extract.

In another aspect, the present invention provides a composition for anti-aging containing one or more herbal extracts selected from the group consisting of pogongyoung, horse riding, powdered herbs and gobon extract as an active ingredient.

The composition containing one or more herbal extracts selected from the group consisting of pogongyoung, horse riding, powder and herbal extracts according to the present invention healed scratch wounds, iNOS, IL-1β and TNF which are inflammation-related factors in LPS-induced cell line model mRNA levels of -α were significantly reduced and anti-inflammatory or wound healing effects were shown by increasing expression of keratinocyte growth factor receptor genes. In addition, the composition according to the present invention inhibits the increase of NF-κB activity and the increase of MMP-1 production by UV, and also inhibits the increase of MMP-1 production with aging and anti-aging effect by increasing the expression of detoxifying genes Indicated. Therefore, pogongyoung, horse riding, powder and extract extract according to the present invention can provide excellent anti-inflammatory, wound healing and anti-aging effect by containing as an active ingredient in the cosmetic composition or pharmaceutical composition.

The present invention provides an anti-inflammatory composition containing 0.0001 to 15% by weight of one or more herbal extracts selected from the group consisting of pogongyoung, horse riding, mountain powder and gobon extract based on the total weight of the composition.

In another aspect, the present invention provides a composition for wound healing containing 0.0001 to 15% by weight relative to the total weight of one or more herbal extracts selected from the group consisting of pogongyoung, horse riding, powdered and kobon extract.

In another aspect, the present invention provides an anti-aging composition containing 0.0001 to 15% by weight relative to the total weight of the composition of one or more herbal extracts selected from the group consisting of pogongyoung, horse riding, powder and gobon extract.

Hereinafter, the present invention will be described in detail.

The pogongyoung, horse riding, powder and herbal extracts used in the present invention can be obtained by extracting each herbal medicine with water or an organic solvent. The organic solvent used in the present invention may be one or more selected from the group consisting of ethanol, methanol, butanol, ether, ethyl acetate and chloroform, or a mixed solvent of these organic solvents and water may be used. Preferably 80% ethanol can be used.

The external preparation composition for skin according to the present invention comprises at least one medicinal herb extract selected from the group consisting of pogongyoung, horseback riding, medicinal herb and medicinal extract extracted by the above method as an active ingredient in an amount of 0.0001 to 15% by weight based on the total weight of the composition. It contains. When the content of the extract is less than 0.0001% by weight, anti-inflammatory, wound healing, anti-aging effects, etc. may not be obtained by the extract. When the content of the extract is more than 15% by weight, the increase in effect is not large compared to the increase in content. The composition according to the present invention may be used alone or in combination of two or more of the pogongyoung, horse riding, acid and extract extract, the effect was more excellent when used in combination of two or more than the extract alone. Preferably, the pogongyoung, horse riding, powder and extract extract is used in a weight ratio of 1: 1: 1: 1, but can be used by mixing in an appropriate ratio in the formulation without particular limitation.

As used herein, the terms "skin damage" or "wound" can be used interchangeably and include, for example, scratches, cuts, torn wounds, pressed wounds, compression wounds, stretch damage, bite wounds, abrasions, Bullet wounds, explosion damage, body piercings, puncture wounds, burns, wind wounds, sun burns, chemical burns, surgical wounds, surgical interventions, medical interventions, host rejection after transplantation of cells, tissues or organs, drug effects, Pharmaceutical side effects, bedsores, radiation damage, skin wounds caused by cosmetics, developmental processes, maturation processes (such as acne), genetic abnormalities, developmental abnormalities and environmental damage.

The anti-inflammatory, wound healing or anti-aging composition according to the present invention may be prepared in the form of a pharmaceutical composition comprising an effective amount of pogongyoung, horse riding, powder and extract extracts, one or more commonly used in the art The above non-toxic, pharmaceutically acceptable carrier, adjuvant or diluent or other active ingredient may be included.

The compositions according to the invention can also be formulated in the form of external preparations for the skin by known methods using pharmaceutically acceptable carriers and excipients.

In addition, the composition according to the present invention may be in the form of a solution, a suspension or an emulsion in an oil or an aqueous medium, or in the form of a dry powder that is dissolved in sterile, pyrogen-free water before use, and may be formulated into an external skin formulation. have. Water-in-oil emulsions are derived from vegetable oils such as olive oil or mineral oils such as liquid paraffin, and are derived from natural phospholipids such as soy bean lecithin and esters of anhydrous hecitol or fatty acids such as sorbitan monooleate. The active ingredient is emulsified using compounds obtained by condensation of ethylene oxide with partial esters derived from anhydrous hexitol anhydride and fatty acids, such as lyoxyethylene sorbitol monooleate.

When the external composition for skin of the present invention is formulated as a cosmetic, it is not particularly limited in the formulation, it is a softening longevity, astringent longevity, nourishing longevity, eye cream, nutrition cream, massage cream, cleansing cream, cleansing foam, cleansing water, It may have a formulation such as powder, essence or pack.

Hereinafter, the present invention will be described in more detail based on the following Examples and Test Examples. However, these examples and test examples are only for helping the understanding of the present invention, the scope of the present invention is not limited to these examples.

Example 1 Preparation of Herbal Medicine Extract

5 L of 80% ethanol aqueous solution was added to 1 kg of dry pogongyoung, horseback riding, powdered acid and high bone, and refluxed three times, followed by immersion at 15 ° C for 1 day. Thereafter, the residue and the filtrate were separated through filter cloth filtration and centrifugation. The separated filtrate was concentrated under reduced pressure to obtain pogongyoung, horse riding, powdered acid and kobon extract, respectively.

Test Example 1 Wound Healing Activity through Visual Form

HaCaT cells, which are human keratinocytes, were distributed and used by Korean Cell Line Bank (Seoul, Korea). HaCaT cells were injected into DMEM medium containing 10% (v / v) FBS, 100 U / ml penicillin and 100 μg / ml of streptomycin and cultured in an animal cell incubator with 37 ° C., 5% CO ₂ feed conditions. HaCaT cells prepared at a concentration of 1.5 × 10 ⁶ per well were incubated for 24 hours to form cell monolayers, and then HaCaT cell monolayers were “scratched-damaged” with a p200 pipette tip. In the culture medium in which the "scratch damaged" cell layer was treated at 10 ppm concentrations of pogongyoung, horse riding, powdered acid and medicated extract prepared in Example 1, respectively. After culturing for 24 hours, the degree of recovery of the scratches was confirmed, and the results are shown in FIG. 1. As a negative control, the same amount of DMSO used to dissolve the herbal extracts was treated.

In addition, in order to confirm the change by the herbal medicine composite formulation, poongyoung, horseback riding, powdered powder and kobon extract prepared in Example 1 was prepared in a composite extract mixed in a weight ratio of 1: 1: 1: 1. In the same manner as above, after the incubation for 24 hours in a culture medium treated with a complex extract of the "scratch damaged" cell layer 10ppm concentration was confirmed the extent of the scratches recovered, the results are shown in FIG.

In the results of FIG. 1, the "scratch damage" of the cells treated with the negative control remained relatively untreated after 24 hours, whereas in the cells treated with the herbal extracts prepared in Example 1 The movement healed the “scratch-damage” and reduced the scratched area after 24 hours. In particular, the healing effect in the cells treated with the composite extract was better than when using the extract alone.

Test Example 2 Anti-inflammatory and Keratin Growth Factor Receptor and Detoxification Enzyme Induction Effect

HaCaT cells, which are human keratinocytes, were distributed and used by Korean Cell Line Bank (Seoul, Korea). HaCaT cells were injected into DMEM medium containing 10% (v / v) FBS, 100 U / ml penicillin and 100 μg / ml of streptomycin and cultured in an animal cell incubator with 37 ° C., 5% CO ₂ feed conditions. HaCaT cells prepared at a concentration of 1.5 × 10 ⁶ per well were adapted for 3 hours with medium without FBS. Thereafter, the pogongyoung, horse riding, powder and kobon extracts prepared in Example 1 and the complex extracts mixed at a weight ratio of 1: 1: 1: 1 were pretreated at 10 ppm for 2 hours and LPS at 1 μg / ml. Addition was further incubated for 8 hours. Total RNA was extracted using TRIzol ^™ (GIBCO BRL, MD, USA) and stored at −80 ° C.

1 μg total RNA was placed in 25 μl of reverse transcription reaction buffer containing 50 mM Tris-HCl, pH 8.3, 75 mM KCl, 3 mM MgCl ₂ , 0.1 M DTT, 10 mM dNTP and 40 units / μl RNase inhibitor, and 1 μg / μl oligo (dT) ₁₆ primer and 200 units of SuperScript II (GiboBRL) reverse transcriptase were added and reacted at 42 ° C. for 1 hour, followed by 2.5 μl of reverse transcription reaction solution. (AmpliTaq) DNA polymerase [0.04U, Perkin Elmer, Shelton, CT], 50 mM Tris, pH 8.3, 0.25 mg / ml bovine serum albumin, 3 mM MgCl ₂ , 0.25 mM dNTPs, SYBR Mix 50 L of PCR reaction buffer containing 1 / 50,000 dilution of Green I (Molecular Probes, Eugene, OR), add 10 μM primer, denature at 94 ° C for 30 seconds, at 53 ° C. Thirty cycles of annealing for 30 seconds and expansion for 1 minute at 72 ° C. were performed.

Relative mRNA levels were analyzed by measuring SYBR Green I fluorescence changes using ICycler software. Each primer was prepared on the basis of published SCI papers by analyzing materials having wound healing efficacy, inflammation inhibitory function, and detoxification activity, and each base sequence is shown in Table 1 below. GAPDH (glyceraldehyde 3-phosphate dehydrogenase) was used as an internal standard to correct quantitative expression levels of genes.

Primer sequence used for PCR Gene / NCBI Gene ID Primer sequence Sequence number Amplification Product Size (bp) PCR reaction temperature (℃) denaturalization Annealing expansion GAPDH / 2597 5'-ATC CCA TCA CCA TCT TCC AG-3 ' One 579 94 58 72 5'-CCT GCT TCA CCA CCT TCT TG -3 ' 2 iNOS / 4843 5'-ATG TCC GAA GCA AAC ATC AC -3 ' 3 401 94 58 72 5'-TAA TGT CCA GGA AGT AGG TG -3 ' 4 IL-1β / 3553 5'-TGC AGA GTT CCC CAA CTG GTA CAT C -3 ' 5 387 94 58 72 5'-GTG CTG CCT AAT GTC CCC TTG AAT C -3 ' 6 TNF-α / 7124 5'-CCT GTA GCC CAC GTC GTA GC -3 ' 7 374 94 58 72 5'-TTG ACC TCA GCG CTG AGT TG -3 ' 8 FGFR2-IIIb / 2263 5'-ACT CGG GGA TAA ATA GTT CCA A-3 ' 9 357 94 60 70 5'-CCT TAC ATA TAT ATT CCC CAG CAT-3 ' 10 NQO1 / 1728 5'-TGA AGG ACC CTG CGA ACT TTC-3 ' 11 185 95 61 70 5'-GAA CAC TCG CTC AAA CCA GC-3 ' 12 GSTP1 / 2950 5'-AGG ACC TCC GCT GCA AAT AC-3 ' 13 105 95 60 70 5'-GGG TCT CAA AAG GCT TCA GTT G-3 ' 14 PRDX1 / 5052 5'-CGG AGA TCA TTG CTT TCA GTG A-3 ' 15 113 95 60 70 5'-AGG TGT ATT GAC CCA TGC TAG AT-3 ' 16

In the present invention, RT-PCR analysis was performed to investigate the effects of herbal extracts on the expression of iNOS, a factor associated with proinflammatory mediator NO, and the expression of IL-1β and TNF-α, another inflammation-related factor. The results are shown in Figures 2a to 2c. Each PCR product was quantified for GAPDH. One-way ANOVA followed by Tukey HSD post hoc test showed significant levels of LPS + compared to ** P <0.01 and * P <0.05. iNOS, IL-1β and TNF-α are inflammation-related factors that play an important role in the inflammatory phase during wound healing and are frequently used to confirm the efficacy of inflammation in many experimental papers. iNOS, IL-1β and TNF-α are genes that are expressed in various ways in the inflammatory response induced by lipopolysaccharide (LPS).

Referring to the results of FIGS. 2A to 2C, LPS-treated cells were overexpressed in respective genes, compared to cells not treated with lipopolysaccharide (LPS), and the treatment of the herbal extracts was iNOS, IL-1β, and TNF-α. Gene expression was strongly inhibited. In addition, the herbal herbal extracts showed better efficacy than when used alone.

go Z, Kopasz N, Szeg C, Pivarcsi A, Koreck A, Dobozy A, Kem

y L, Sz

ll M. The expression of keratinocyte growth factor receptor (FGFR2-Ⅲb) correlates with the high proliferative rate of HaCaT keratinocytes. Exp Dermatol. 2006 Aug;15(8):596-605). 실시예 1에서 제조한 포공영, 승마, 산약 및 고본 단독 추출물과 이들을 1:1:1:1의 중량비로 혼합한 복합 추출물에 의한 각질형성세포 성장인자 수용체(FGFR2-Ⅲb) 유전자의 발현 유도 효과를 측정한 결과를 도 3에 나타내었다. 도 3의 결과에서, 실시예 1에서 제조한 포공영, 승마, 산약 및 고본 추출물 및 이들의 복합 추출물이 모두 각질형성세포 성장인자 수용체 유전자의 발현을 유도하였으며, 특히 단독 추출물보다는 이들의 혼합한 복합 추출물에 의한 각질형성세포 성장인자 수용체(FGFR2-Ⅲb) 유전자의 발현 유도 효과가 더 높았다.In addition, expression of keratinocyte growth factor receptor (FGFR2-IIIb) genes has been reported to correlate with cellular proliferative effects (Nagy N, Bata-Cs).

go Z, Kopasz N, Szeg C, Pivarcsi A, Koreck A, Dobozy A, Kem

y L, Sz

ll M. The expression of keratinocyte growth factor receptor (FGFR2-IIIb) correlates with the high proliferative rate of HaCaT keratinocytes. Exp Dermatol. 2006 Aug; 15 (8): 596-605) . Induction effect of the keratinocyte growth factor receptor (FGFR2-IIIb) gene by the extract of pogongyoung, horse riding, powder and kobon alone extract prepared in Example 1 and a complex extract mixed with a weight ratio of 1: 1: 1: 1 The measured result is shown in FIG. In the results of Figure 3, pogongyoung, horse riding, powder and extract extracts and their complex extracts prepared in Example 1 all induced the expression of keratinocyte growth factor receptor genes, in particular the complex extracts thereof mixed rather than the extract alone Expression of keratinocyte growth factor receptor (FGFR2-IIIb) gene was higher.

It has also been reported that expression of detoxification genes such as GSTs, NQO1 and PRDX1 induces cellular protective action from oxidative damage (Xie C, Lovell MA, Xiong S, Kindy MS, Guo J, Xie J, Amaranth V, Montine TJ , Markesbery WR.Expression of glutathione-S-transferase isozyme in the SY5Y neuroblastoma cell line increases resistance to oxidative stress.Free Radic Biol Med. 2001 Jul; 31 (1): 73-81; Liu Y, Kern JT, Walker JR, Johnson JA, Schultz PG, Luesch H. A genomic screen for activators of the antioxidant response element.Proc Natl Acad Sci US A. 2007 Mar; 104 (12): 5205-10; Carcinogenesis. 2000 May; 21 (5): 1013 -6.Induction of murine intestinal and hepatic peroxiredoxin MSP23 by dietary butylated hydroxyanisole.Ishii T, Itoh K, Akasaka J, Yanagawa T, Takahashi S, Yoshida H, Bannai S, Yamamoto M). 4a to 4c show the effect of inducing the expression of GSTs, NQO1 and PRDX1, which are the detoxification genes of the pogongyoung, horse riding, powder and kobon extracts prepared in Example 1 and the complex extracts mixed at a weight ratio of 1: 1: 1: 1 Is the result of measurement. The pogongyoung, horse riding, powder and extract extract and the combination extract thereof prepared in Example 1 all induced the expression of the detoxification genes GSTs, NQO1 and PRDX1, in particular GSTs, NQO1 and The expression inducing effect of PRDX1 was higher.

Test Example 3 Inhibitory Effect of UV on NF-κB Activity Increase

HaCaT cells, a human keratinocyte cell line, were cultured in 100 mm cell culture dishes at 10 ⁶ densities, and UVB was irradiated using FS40 lamps (Westinghouse, Pittsburgh, P., USA). After irradiating at 10mJ / cm 2, pogongyoung, horseback riding, powdering and gobon extract prepared in Example 1 was replaced with a medium containing 10ppm each.

After 6-24 hours of culture, the nuclear extract was extracted using a TransFactor extraction kit (BD Bioscience Clontech, San Jose, C.A., USA (BD Bioscience Clontech, San Jose, CA, USA)). NF-κB activity was measured using a transfactor kit (BD Bioscience Clontech, San Jose, CA, USA), and the results are shown in FIG. 5.

In the results of FIG. 5, the NF-κB activity was greatly increased by ultraviolet irradiation at first, but the increase of NF-κB activity by ultraviolet rays was suppressed after the herbal extract of Example 1 was treated.

Test Example 4 Effect of Inhibiting the Increase of MMP-1 Production by UV

HaCaT cells, a human keratinocyte cell line, were cultured in a 100 mm cell culture dish at a density of 10 ⁶ , and pretreated with poppongyoung, horseback riding, powder and gobon extracts prepared in Example 1 by 10 ppm, respectively, and then FS40 lamps (Westinghouse, Pittsburgh, Ultraviolet B was irradiated at 10 mJ / cm 2 using Pennsylvania, USA. After replacing the pogongyoung, horse riding, powdered acid and kobon extract with a medium containing 10ppm each incubation for 48 hours, the amount of MMP-1 produced was measured by ELISA kit [amersharm pharmacia, cat. #; RPN2610]. Each measured amount of MMP-1 was corrected relative to the total protein, and the results are shown in FIG. 6.

In the results of FIG. 6, MMP-1 production was initially increased by UV irradiation, but the increase in MMP-1 production by UV treatment was significantly suppressed by treating pogongyoung, horse riding, powdered powder and kobon extract prepared in Example 1. .

Test Example 5 Inhibition Effect of MMP-1 Production by Aging

Human dermal fibroblasts were cultured in DMEM medium containing 10% fetal calf serum with or without 10 ppm of pogongyoung, horse riding, powder and kobon extracts prepared in Example 1 (untreated group). When reaching a density of 80 to 90%, the cells were passaged at 1/2 to obtain cells at Passage 5, Passage 10, Passage 20 and Passage 30, and the amount of MMP-1 produced was measured. Indicated.

In the results of Figure 7, pogongyoung, horse riding, powder and extract extract according to the present invention inhibited the increase of MMP-1 production with aging in the proliferating aging cell model.

As described above, pogongyoung, horse riding, powder and herbal extracts used as an active ingredient in the present invention to suppress the inflammation-related factors and promote re-epithelialization to help the progression from the inflammatory phase to the proliferative phase during wound healing and during the growth phase It helped the maturation of connective tissue cells by helping to secrete factors that stimulate cell proliferation and help in wound healing. In addition, by increasing the gene expression of enzymes involved in the detoxification action with an anti-inflammatory effect, it has the effect of inhibiting skin aging by inhibiting skin damage caused by external stimuli such as UV. The combined regimen had a better effect than using these components alone.

Therefore, pogongyoung, horse riding, powder and extract extract according to the present invention shows an effect of shortening the treatment time by little by little as a whole help to the inflammatory and mature phase of the initial process during the overall wound treatment process, the pogongyoung, horse riding, It was possible to provide an anti-inflammatory or wound healing agent containing a powder and an extract as an active ingredient. In addition, the present invention may provide an anti-aging agent containing the pogongyoung, horse riding, powder and extract extract as an active ingredient, and inhibits skin aging by external stimulation such as UV through anti-inflammatory or detoxification action have.

Formulation Example 1 Nutritional Cosmetics

Nutritional longevity was prepared according to the composition described in Table 2 below in a conventional manner.

ingredient Content (% by weight) Purified water Remaining amount glycerin 8.0 Butylene glycol 4.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Carbomer 0.1 Pogongyoung, horse riding, pesticide or herbal extract 0.05 Caprylic / Capric Triglycerides 8.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Cetearyl Alcohol 1.0 antiseptic Quantity incense Quantity Pigment Quantity Triethanolamine 0.1

Formulation Example 2 Nutrition Cream

Nutritional cream was prepared in a conventional manner according to the composition shown in Table 3.

ingredient Content (% by weight) Purified water Remaining amount glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Pogongyoung, horse riding, pesticide or herbal extract 3.0 Caprylic / Capric Triglycerides 3.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 antiseptic Quantity incense Quantity Pigment Quantity Triethanolamine 0.1

Formulation Example 3 Massage Cream

To prepare a massage cream in a conventional manner according to the composition described in Table 4.

ingredient Content (% by weight) Purified water Remaining amount glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 Pogongyoung, horse riding, pesticide or herbal extract 1.0 Caprylic / Capric Triglycerides 3.0 Beeswax 4.0 Cetearyl Glucoside 1.5 Sesqui oleic acid sorbitan 0.9 Vaseline 3.0 antiseptic Quantity incense Quantity Pigment Quantity paraffin 1.5

[Formulation Example 4] Pack

To prepare a pack in a conventional manner according to the composition described in Table 5.

ingredient Content (% by weight) Purified water Remaining amount glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 Pogongyoung, horse riding, pesticide or herbal extract 0.5 Nonyl Phenyl Ether 0.4 Polysorbate 60 1.2 antiseptic Quantity incense Quantity Pigment Quantity ethanol 6.0

Formulation Example 5 Ointment in External Skin Preparation

The ointment was prepared in a conventional manner according to the composition described in Table 6.

ingredient Content (% by weight) Purified water Remaining amount glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 Pogongyoung, horse riding, pesticide or herbal extract 1.0 Caprylic / Capric Triglycerides 3.0 Squalane 1.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Cetearyl Alcohol 1.0 antiseptic Quantity incense Quantity Pigment Quantity Beeswax 4.0

1 is a result of confirming the wound healing effect of the pogongyoung, horse riding, powder and herbal extracts in accordance with the in vitro wound healing assay (in vitro wound healing assay) and the wound healing effect according to the complex prescription.

Figure 2a to 2c is a graph showing the results of measuring the expression level of iNOS, IL-1β and TNF-α mRNAs in HaCaT cells induced inflammation with LPS.

Figure 3 is a graph showing the expression inducing effect of the keratinocyte growth factor receptor (FGFR2-IIIb) gene by pogongyoung, horse riding, powder and extract extract.

Figures 4a to 4c is a result of measuring the change in the expression of GSTP1, NQO1 and PRDX1 gene by pogongyoung, horse riding, powder and kobon extract

Figure 5 is the result of measuring the change in NF-κB activity by pogongyoung, horse riding, powder and gobon extract.

6 is a result confirming the inhibitory effect of the increase in MMP-1 expression by irradiation of pogongyoung, horse riding, powder and gobon extract.

7 is a result confirming the inhibitory effect of increased MMP-1 expression during cell aging according to repeated passage cultures of pogongyoung, horse riding, powder and gobon extract.

<110> AMOREPACIFIC CORPORATION <120> Skin external composition containing Taraxacum platycarpum H.          Dahlsi, Cimicifuga heracleifolia, Dioscorea opposita or Angelica          tenuissima Nakai <160> 16 <170> KopatentIn 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for GAPDH <400> 1 atcccatcac catcttccag 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for GAPDH <400> 2 cctgcttcac caccttcttg 20 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for iNOS <400> 3 atgtccgaag caaacatcac 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for iNOS <400> 4 taatgtccag gaagtaggtg 20 <210> 5 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> IL-1 beta <400> 5 tgcagagttc cccaactggt acatc 25 <210> 6 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer for IL-1 beta <400> 6 gtgctgccta atgtcccctt gaatc 25 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for TNF-alpha <400> 7 cctgtagccc acgtcgtagc 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for TNF-alpha <400> 8 ttgacctcag cgctgagttg 20 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer for FGFR2-IIIb <400> 9 actcggggat aaatagttcc aa 22 <210> 10 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer for FGFR2-IIIb <400> 10 ccttacatat atattcccca gcat 24 <210> 11 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer for NQO1 <400> 11 tgaaggaccc tgcgaacttt c 21 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for NQO1 <400> 12 gaacactcgc tcaaaccagc 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer for GSTP1 <400> 13 aggacctccg ctgcaaatac 20 <210> 14 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer for GSTP1 <400> 14 gggtctcaaa aggcttcagt tg 22 <210> 15 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer for PRDX1 <400> 15 cggagatcat tgctttcagt ga 22 <210> 16 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> primer for PRDX1 <400> 16 aggtgtattg acccatgcta gat 23  

Claims (4)

Pogongyoung, horse riding, powdered herbal and anti-inflammatory skin external composition comprising as an active ingredient one or more herbal extracts selected from the group consisting of extracts. Pogongyoung, horse riding, powdered medicine and herbal extracts for wound healing skin composition containing as an active ingredient at least one herbal extract selected from the group consisting of extract. Anti-aging skin composition for external aging containing at least one herbal extract selected from the group consisting of pogongyoung, horse riding, powdered acid and kobon extract as an active ingredient. The composition for external application for skin according to any one of claims 1 to 3, wherein the herbal extract is contained in an amount of 0.0001 to 15% by weight based on the total weight of the composition.

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