Composition for preventing or treating atopic dermatitis, containing placenta extract

Abstract

When the human placenta extract of the present invention is applied to cells, animals and humans, atopic dermatitis can be prevented or treated, and more particularly, Th1 / Th2 imbalance caused by an allergic immune response considered as a cause of atopic dermatitis can be regulated by administering the placenta extract of the present invention, and the human placenta extract of the present invention can be applied to cells, animals and humans. The placenta extract according to the present invention is confirmed to inhibit the production of IgE, which is the biggest characteristic of atopic dermatitis, thereby being able to prevent or treat diseases.

Description

Technical Field

[0001] The present invention relates to a composition for preventing or treating atopic dermatitis containing human placenta extract (HP E or PE) as an active ingredient. More specifically, it relates to a composition having the effect of restoring the disrupted Th1 / Th2 balance in spleen cells of mice with Th2 overreaction, or a pharmaceutical composition having the effect of improving atopic dermatitis symptoms and inhibiting IgE production in an atopic dermatitis animal model. Background Art

[0002] Atopic dermatitis is a chronic, recurring inflammatory disease of the skin that precedes asthma and allergic diseases (Boguniewicz & Leung, 2010). While its etiology remains unclear, it is hypothesized to be caused by a variety of factors, including environmental and genetic factors, immune imbalances, and abnormalities in the skin barrier (Larsen & Hanifin, 2002; Searing & Leung, 2010). Furthermore, atopic dermatitis is characterized by primary skin lesions, such as severe itching, erythema, edema, exudation, dry skin, crusting, and scaling, and secondary physical symptoms, such as sleep disturbances, decreased concentration, and mood swings (Kader et al., 2021). The epidermis, the outermost layer of the skin, performs a barrier function that protects the body from various external stimuli. Its homeostasis is maintained through the repetitive processes of epidermal cell formation, differentiation, and shedding (Benedetto et al., 2012). Keratinocytes, the representative cells of the epidermis, form the skin barrier and, when exposed to stressful environments, participate in inflammatory and immune responses. However, when persistent skin damage chronically induces inflammation, it can lead to dry skin and skin diseases such as atopic dermatitis (Hudson, 2006; Natsuga, 2014). Atopic dermatitis involves multiple cells and factors, including cytokines expressed by T lymphocytes and Th1 / Th2 cells, and antigen-specific immunoglobulin E (IgE) expressed by dendritic cells (Kim et al., 2012). Normally, Th1 / Th2 cells maintain a balance and maintain immune responses. However, in atopic dermatitis, the conversion of T lymphocytes to Th2 cells is promoted, resulting in increased secretion of inflammatory cytokines by these cells and an increase in IgE in the blood, further amplifying the inflammatory response. Therefore, in atopic dermatitis, the immune response driven by Th2 cells predominates. It is well known that thymus and activation-regulated chemokine (TARC / CCL17) and macrophage-derived chemokine (MDC / CCL22) induce the migration of Th2 lymphocytes to inflammatory sites and are important chemokines for inflammatory skin diseases such as atopic dermatitis that are sensitive to Th2 cells (Rozyk et al., 2005).Furthermore, TARC and MDC belong to the C-C motif chemokine receptor 4, which is produced in various cells, including skin keratinocytes (Saeki & Tamaki, 2006). TARC and MDC are significantly elevated in the serum of patients with atopic dermatitis and have been reported to be associated with atopic dermatitis (Nakazato et al., 2008). Furthermore, in atopic dermatitis, the chemokine regulated upon activation (RANTES / CCL5) is involved in the migration and activity of eosinophils and T lymphocytes, and lymphocytes increase their adhesion to endothelial cells, inducing inflammation (Luster, 2002). Therefore, reducing the production of RANTES, TARC, and MDC in skin keratinocytes could have a therapeutic effect in inflammatory skin diseases such as atopic dermatitis. Existing research indicates that in skin keratinocytes, Th2 chemokine expression induced by tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) is downregulated by inhibiting the activity of mitogen-activated protein kinase (MAPK) / P38, the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway, and signal transducer and activator of transcription (STAT1) (Park et al., 2015). Furthermore, few studies have reported on the use of various traditional Chinese medicines or natural products to regulate atopic dermatitis (Cho, 2012; Song & Kim, 2009; Yun, 2012), and relevant research remains insufficient.

[0003] Currently, most allergy treatments, including atopic dermatitis, use steroids, which can cause serious side effects with long-term use. In particular, since atopic dermatitis patients are mostly children and adolescents, the development of safe, non-toxic therapeutic agents is crucial. While research is underway to overcome these limitations, the development of therapeutic agents remains insufficient.

[0004] DNCB, which is the most commonly used substance in atopic dermatitis animal experiments, is a representative substance that induces allergic contact dermatitis, causing an atopic cellular immune response and leading to an increase in mast cells and T lymphocytes.

[0005] Human placenta extract (HPE) contains a variety of growth factors, cytokines, and other physiologically active substances and is widely used for fatigue relief and antioxidant purposes (Lee KK, et al., Evid Based Complement Alternat. Med., vol. 2012, (2012) p. 130875). Furthermore, it is known that HPE has been shown to induce liver regeneration and improve liver function in animal studies, and to influence wound healing through the production of TGF-β and VEGF. However, despite the great interest in HPE, its functions remain largely unstudied. In particular, studies on inflammatory markers associated with atopic dermatitis have never been conducted. Summary of the Invention

[0006] Technical issues

[0007] The present invention attempts to develop a composition for the prevention or treatment of atopic dermatitis containing a human placenta extract as an active ingredient. More specifically, the present invention attempts to develop a composition that has the effect of regulating the Th1 / Th2 imbalance caused by Th2 excessive response or a pharmaceutical composition that has the effect of improving atopic dermatitis symptoms and inhibiting IgE production in an atopic dermatitis animal model.

[0008] Solutions to the Problem

[0009] 1. A composition for preventing or treating atopic dermatitis comprising a human placenta extract as an active ingredient.

[0010] 2. A composition for inhibiting the production of TARC or RANTES produced by an allergic immune response, comprising a human placenta extract as an active ingredient.

[0011] 3. A composition for increasing the production of TGF-β1, a Treg cytokine that is an immunomodulatory indicator, comprising a human placenta extract as an active ingredient.

[0012] 4. The composition according to item 1 above, wherein the composition inhibits the production of TARC or RANTES produced by an allergic immune response.

[0013] 5. The composition according to item 1 above, wherein the composition increases the production of TGF-β1, a Treg cytokine that is an immunoregulatory indicator.

[0014] 6. The composition according to item 1 above, wherein the composition increases the decreased Th1 cytokine / Th2 cytokine ratio.

[0015] 7. The composition according to item 1 above, characterized in that the composition inhibits the production of IgE in the blood.

[0016] 8. The composition according to item 1 above, characterized in that the composition reduces dermatitis scoring in atopic dermatitis symptoms.

[0017] 9. The composition according to item 1 above, characterized in that the extract is a human placenta extract prepared by defatting with acetone and then hydrolyzing with pepsin and hydrochloric acid.

[0018] 10. The composition according to 2 above, characterized in that it is a human placenta extract prepared by defatting with acetone and then hydrolyzing with pepsin and hydrochloric acid.

[0019] 11. The composition according to 3 above, characterized in that it is a human placenta extract prepared by defatting with acetone and then hydrolyzing with pepsin and hydrochloric acid.

[0020] 12. In the pharmaceutical composition of the above 1, it is characterized in that the dosage form of the above composition is one selected from the group consisting of injection, cream, gel, patch, spray, ointment, plaster, lotion, microneedle, roller and their combination.

[0021] 13. In the pharmaceutical composition of the above 2, it is characterized in that the dosage form of the above composition is one selected from the group consisting of injection, cream, gel, patch, spray, ointment, plaster, lotion, microneedle, roller and their combination.

[0022] 14. In the pharmaceutical composition of the above 3, it is characterized in that the dosage form of the above composition is one selected from the group consisting of injection, cream, gel, patch, spray, ointment, plaster, lotion, microneedle, roller and their combination.

[0023] 15. A food composition for improving atopic dermatitis comprising, as an active ingredient, a human placenta extract prepared by acetone defatting followed by hydrolysis with pepsin and hydrochloric acid.

[0024] 16. In the food composition of 15 above, it is characterized in that the food composition has a dosage form selected from the group consisting of functional food, nutritional supplements, health food, food additives, feed and combinations thereof.

[0025] Effects of the Invention

[0026] When the human placental extract of the present invention is applied, atopic dermatitis can be prevented or treated. More specifically, the imbalance of Th1 / Th2 caused by excessive Th2 reaction can be adjusted. In addition, by confirming that the placental extract of the present invention inhibits the production of IgE, which is the most characteristic feature of atopic dermatitis, the disease can be prevented or treated. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This graph shows the results of confirming the cell viability by CCK-8 analysis when the placenta extract (PE) of the present invention was administered to skin keratinocytes.

[0028] FIG. 2 is a graph showing the results of confirming the therapeutic efficacy of the placenta extract (PE) of the present invention on atopic dermatitis by qRT-PCR after administration to skin keratinocytes.

[0029] Figure 3 This figure shows the results of an ex-vivo experiment confirming the therapeutic efficacy of the present invention on atopic dermatitis by treating mouse spleen cells biased toward Th2 cell response with the placenta extract (PE) of the present invention.

[0030] FIG4 is a graph showing the results of visual observation of the degree of improvement of atopic dermatitis after administration of the placenta extracts (PE200 and PE400) of the present invention to an atopic dermatitis-induced mouse model.

[0031] Figure 5 The graph shows the results of confirming the degree of improvement of atopic dermatitis using spleen numerical values ​​after administration of the placenta extracts (PE200 and PE400) of the present invention to an atopic dermatitis-induced mouse model.

[0032] Figure 6 This figure shows the results of confirming the degree of side effects of the samples by the amount of muscle mass loss after administration of the placenta extracts (PE200 and PE400) of the present invention to an atopic dermatitis-induced mouse model.

[0033] Figure 7 The graph shows the results of confirming the degree of improvement of atopic dermatitis by serum IgE levels after administration of the placenta extracts (PE200 and PE400) of the present invention to an atopic dermatitis-induced mouse model. DETAILED DESCRIPTION

[0034] The composition of the present invention can be topically administered to the site of atopic dermatitis or to the site of skin inflammation for which treatment is desired. Specifically, the composition of the present invention can be delivered by any method that selectively delivers an effective amount of the composition to the skin site in need of treatment. For example, the administration can be achieved by direct topical administration (e.g., applying the composition of the present invention to the skin of the treatment site in the form of a lotion or gel, or spraying the composition of the present invention onto the treatment site in the form of a liquid) to the treatment site, or by injection (e.g., subcutaneous or intradermal injection) into the treatment site comprising the skin after formulation using an injectable formulation.

[0035] According to one embodiment of the present invention, the treatment can be performed by directly topically administering a composition containing other atopic dermatitis therapeutic agents to the treatment site and injecting a composition containing placenta extract to the treatment site.

[0036] Therefore, the present invention provides a pharmaceutical preparation comprising the above composition.

[0037] The pharmaceutical preparation according to the present invention may be in the form of an injection or a skin external preparation, for example, may be in a form selected from the group consisting of an injection, cream, gel, patch, spray, ointment, plaster, lotion, microneedle, roller and a combination thereof.

[0038] According to one embodiment of the present invention, the composition of the present invention can be formulated in the form of an injection for subcutaneous injection into the treatment area. In this case, it may further comprise an excipient suitable for injection. Preferably, the composition comprising human placenta extract can be formulated in the form of an injection. The above-mentioned suitable excipients may include saline, bacteriostatic saline, sterile water, etc., but are not limited thereto. The preparation method of this injection can be easily prepared according to methods well known in the art.

[0039] Furthermore, the composition of the present invention can be formulated in the form of a skin external preparation suitable for direct administration to the treatment site. In this case, it may further comprise a physiologically acceptable carrier or diluent suitable for topical administration to the skin. Preferably, a composition comprising other atopic dermatitis therapeutic agents can be formulated in the form of a skin external preparation. The above-mentioned physiologically acceptable carrier or diluent may include water, physiological saline, creams, lotions, various forms of gels, and short-chain alcohols and glycols (such as ethanol and propylene glycol), but is not limited thereto.

[0040] When the preparation according to the present invention is in the form of an external preparation for skin, it may further contain an ingredient selected from the group consisting of a sunscreen, a skin conditioner, a skin protectant, an emollient, a moisturizer, and a mixture thereof.

[0041] In the present invention, the placenta extract is administered in an amount of 0.5 to 3 bottles (2 ml / vial) over 1 to 2 weeks, preferably 1 to 3 bottles at a time. However, when actually applicable to the patient, the amount is not limited and can be appropriately increased or decreased for administration.

[0042] Preparation Example

[0043] The placenta extract of the present invention can be prepared by the methods exemplified below, but is not limited to the methods described herein.

[0044] The placental extract of the present invention can be prepared by first defatting the placenta (human placenta) by acetone treatment, and then fully hydrolyzing it with pepsin and hydrochloric acid to prevent the formation of incomplete hydrolyzates. In order to apply the placental extract of the present invention to experimental cells and animals (including humans), it is prepared as an injectable dosage form and diluted in a minimal culture medium at a v / v% ratio. In this case, the culture medium is Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 1% penicillin.

[0045] Example

[0046] Hereinafter, the present invention will be described in more detail by way of the following examples. However, the following examples are merely illustrative of the present invention, and the scope of the present invention is not limited to these examples.

[0047] 1. Culture of Human Skin Keratinocytes

[0048] HaCaT (human keratinocyte cell line), a type of human skin keratinocyte, was cultured in DMEM (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin in an incubator at 37°C and 5% CO2. Subculture was performed when the HaCaT cells occupied 70-80% of the culture dish.

[0049] 2. Survival Assay for Evaluating Cytotoxicity

[0050] In order to confirm the cytotoxicity of the placental extract (PE) as a sample of the present invention on HaCaT cells, a cell viability experiment was performed using the CCK-8 assay. The number of HaCaT cells was counted and aliquoted into a 96-well plate to form 5×10 4cells / well, cultured in an incubator at 37°C and 5% CO2 for 24 hours. Afterwards, for each attached cell, PE was selected to be 0.625%, 1.25%, 2.5%, 5%, 10%, 20%, and 40% for treatment, and, as a positive control group, dexamethasone (dexamethason e), the most commonly used steroid drug in atopic dermatitis, was selected, and dexamethasone 0.1μM and 1μM were treated for 24 hours respectively. After the above treatment, in order to determine the survival rate of the cells, the supernatant was removed, and then 10% CCK-8 solution was mixed with culture medium without FBS added, and 100μl was treated per well. After reacting at room temperature for 30 minutes, the absorbance was measured at 450nm using a microplate reader (Molecular Devices, USA). The cell survival rate (%) was calculated according to the following formula.

[0051]

[0052] AC: absorbance of the control group

[0053] AS: absorbance of the sample

[0054] like Figure 1 As shown, it was confirmed that PE of the present invention did not cause cytotoxicity to HaCaT cells at all concentrations. Furthermore, the positive control dexamethasone showed no cytotoxicity at 0.1 μM and 1 μM, so 1 μM was used in all subsequent experiments.

[0055] 3. Evaluation of anti-atopic dermatitis efficacy in human skin keratinocytes

[0056] In order to evaluate the efficacy of improving atopic dermatitis, the degree of inhibition of the production of TARC and RANTES, which are allergic immune indicators, and the amount of TGF-β1 produced by Tre g cytokine, which is an immune regulation indicator, were confirmed by PE of the present invention. HaCaT cells (1×10 5cells / well) were dispensed into a 24-well plate and cultured in an incubator at 37°C and 5% CO2 for 24 hours. Afterwards, a culture medium containing TNF-α+IFN-γ (10 ng / mL each) and PE (5%, 10%, 20%, and 40%, respectively) were simultaneously applied to each attached cell and cultured in an incubator at 37°C and 5% CO2 for 24 hours. The cells were washed twice with PBS, and RNA was isolated using an RNeasy mini kit (Qiagen) according to the manufacturer's protocol. After quantification using the isolated RNA, cDNA was synthesized using a High capacity RNA to cDNA kit (Thermo Fisher Scientific) according to the manufacturer's protocol. Afterwards, 0.5 μl of each of cDNA (10 ng), SYBR Green 10 μL, and 10 μM forward / reverse primers were mixed with RNase-free water (7 μl) to prepare the PCR sample. The reaction was then performed in a qPCR device (Quant Studio 3; Thermo Fisher Scientific) at 95°C for 15 seconds, 55°C for 30 seconds, and 72°C for 30 seconds for 40 cycles. The primer sequences used are shown in Table 1 below. Dexamethasone 1 μM, the most commonly prescribed steroid drug for atopic dermatitis as an injection or topical agent, was used as a positive control.

[0057] Table 1

[0058]

[0059]

[0060] like Figure 2a 、 Figure 2b and Figure 2cAs shown, it was confirmed that when HaCaT cells, which are skin keratinocytes, were treated with TNF-α+INF-γ (10 ng / mL each), which are atopic dermatitis inducing substances (referred to as the control group), the expression levels of TARC and RANTES, which are allergic immune indicators, in the control group were significantly increased, and the expression level of TGF-β1, which is an immunomodulatory indicator, was significantly decreased (p<0.001) compared to the normal group that was not treated with anything. It was confirmed that when the PE of the present invention was treated at different concentrations (5-40%) with cells inducing atopic dermatitis, the expression levels of mRNA in the case of TARC and RANTES were statistically significantly reduced in a concentration-dependent manner (p<0.001). Figure 2a and Figure 2b ), in the presence of TGF-β1, the concentration-dependent increase ( Figure 2c ). It was confirmed that the mRNA expression of TARC was also reduced in the positive control group of dexamethasone 1 μM, while the expression of TGF-β1 was increased. Therefore, this experiment confirmed that the PE of the present invention is effective in improving the symptoms of atopic dermatitis by reducing the production of allergic immune factors (TARC or RANTES) and increasing the expression of the immune regulatory factor TGF-β1.

[0061] 4. Animal husbandry and experimental group classification

[0062] 4-1) Ex-vivo

[0063] Six-week-old female Balb / c mice were purchased and acclimated for one week before use. During the experiment, feed and water were freely available. The temperature in the housing room was maintained at 22 ± 2°C, the humidity at 50 ± 20%, and the light-dark cycle was maintained at 12 h (12 h light / 12 h dark).

[0064] 4-2) In-vivo

[0065] Six-week-old male Balb / c mice were purchased as experimental animals and, after a one-week acclimation period, were categorized as shown in Table 2 below. During the experiment, feed and water were freely available. The temperature in the housing room was maintained at 22±2°C, the humidity at 50±20%, and the light-dark cycle was maintained at 12 hours (12 hours light / 12 hours dark). The animals' food and water intake was measured every other week, as was their body weight change, and their general health was observed.

[0066] Table 2

[0067]

[0068] The composition of Table 2 corresponds to the animal experiments.

[0069] 5. Immunization of Experimental Animals and Isolation of Spleen Cells

[0070] In order to confirm the degree of immunomodulatory effect of PE as a sample of the present invention on splenocytes other than RBCs in spleen tissue (splenocyte), an ex vivo test was performed. After mixing 1.5 mL (13 mg / mL) of aluminum hydroxide (Sigma) solution, 10 mg / mL of ovalbumin, and 0.46 mL of PBS, the mixture was allowed to react at room temperature for 20 minutes. Afterwards, 0.2 mL (2 mg aluminum hydroxide + 1 mg ovalbumin) was injected intraperitoneally into mice that had undergone the environmental acclimatization process. On the 6th day, 0.2 mL was injected intraperitoneally again for boosting. On the 13th day, the mice were sacrificed, the spleen was removed, and 2×10 splenocytes were obtained from the spleen. 6 Cells were added to a 24-well plate along with 250 μl of ovalbumin (4 mg / mL) and PE (5% or 10%). The cells were cultured for 7 days in RPMI medium containing 10% FBS in a 5% CO2 incubator. After 7 days of culture, the supernatant was collected and used in the experiment. As a positive control, dexamethasone (1 μM) was used, as in the previous experiment.

[0071] 6. Evaluation of the Th1 cytokine / Th2 cytokine ratio in mouse spleen cells

[0072] IL-12 and IL-4 cytokines were evaluated using R&D Systems ELISA kits. Following the manufacturer's protocol, the capture antibody was diluted in coating buffer and 100 μl was added to each well. The cells were incubated at room temperature overnight. After the wash step, 300 μl of reagent diluent was added to each well and the cells were incubated at room temperature for 1 hour for blocking. Subsequently, 100 μl of the standard and the test substance of the present invention, diluted starting from the highest concentration, were added to each well and incubated at room temperature for 2 hours. Next, 100 μl of the detection antibody was added to each well and incubated at room temperature for 2 hours. Then, 100 μl of streptavidin-HRP B was added to each well and incubated at room temperature for 20 minutes, protected from light. Add 100 μl of substrate solution to each well and incubate at room temperature in the dark for 20 minutes. Finally, add 50 μl of stop solution to terminate the reaction. Measure absorbance at 450 nm and 540 nm. The result is calculated by subtracting the 540 nm absorbance from the 450 nm absorbance and applying the standard trendline formula.

[0073] It has been observed that in immune allergic reactions, due to abnormal regulation of T cell function, Th2 cell response is dominant over Th1 cell response, thus leading to an imbalance in the secretion of cytokines in Th1 cells and Th2 cells. Figure 3 As shown, after ovalbumin was administered and the spleen cells of mice biased by Th2 response were treated with the PE of the present invention, the expression levels of IL-12 (Th1 cytokine) and IL-4 (Th2 cytokine) were confirmed, and the IL-12 / IL-4 cytokine ratio was measured. In the control group without any treatment, the IL-12 / IL-4 ratio was less than 1, which is a very low value. On the contrary, when the PE of the present invention was treated, the IL-12 / IL-4 ratio was statistically significantly increased. In particular, it was confirmed that the increase was the largest in the 10% PE treatment group of the present invention ( Figure 3). Furthermore, a significant increase in the IL-12 / IL-4 ratio was also observed in the positive control group treated with 1 μM dexamethasone. Therefore, this experiment confirmed that the PE of the present invention has the effect of restoring the balance of Th1 / Th2 cell responses in mouse spleen cells that have been biased toward Th2 cell responses.

[0074] 7. Induce atopic dermatitis

[0075] On the 6th day of acclimation, all mice used in the experiment were depilated using animal clippers and depilatory cream. After depilation, the normal group and the induced group were separated by body weight and TEWL (Transepidermal water loss) measurement results. 24 hours after depilation, sensitization was performed twice every two weeks using 200 μL of 1% dinitrochlorobenzene (DNC B). DNCB solution was prepared by dissolving DNCB in a solution of acetone: olive oil = 3:1. After one week of inducing atopic dermatitis using 1% DNCB, the treated group was divided into two groups based on the IgE concentration in the blood and visual observation. Afterwards, sensitization was performed three times every three weeks using 200 μL of 0.5% DNCB starting from the second week.

[0076] 8. Visual observation and dermatitis scoring

[0077] Visual evaluation measures the severity of atopic dermatitis symptoms by summing the scores for three of six evaluation items. The evaluation items are erythema, dryness, and scarring. Each item is scored as no symptoms (0 points), mild (1 point), moderate (2 points), or severe (3 points). The total score ranges from 0 to 9.

[0078] In order to confirm the therapeutic efficacy of the placental extract (PE) of the present invention in a mouse model of atopic dermatitis induced by application of DNCB, visual observation of skin lesions and dermatitis scoring were performed after the final administration of the test substance. Figure 4a As shown in the figure, compared with the normal group (N), severe atopic dermatitis skin lesions appeared in the induced group (C), and the degree of atopic dermatitis was alleviated in the PE200 and PE400 of the group administered with the test substance of the present invention and the DEX-administered group as the positive control group. Based on this result, the Dermatitis score was measured and quantified, as shown in the figure. Figure 4bAs shown, compared with the induction group, the symptoms were reduced in the PE200 to PE400 administration groups and significantly recovered in the PE400 and DEX administration groups.

[0079] 9. Analysis of Organ Changes

[0080] 9-1) Spleen index

[0081] After the experiment, Balb / c mice were sacrificed, the spleens were removed, and their weights were measured. The spleen value was calculated by dividing the spleen weight (mg) by the body weight (g).

[0082] 9-2) Muscle mass

[0083] After the experiment, Balb / c mice were sacrificed, and the gastrocnemius and soleus muscles, serving as calf muscles, were removed and weighed. Muscle mass was calculated by dividing the calf muscle weight (mg) by the body weight (g).

[0084] It is well known that when atopic dermatitis occurs, allergic immunity leads to excessive differentiation of Th2 cells, an increase in T lymphocytes, and an enlargement of the spleen. Figure 5 As shown, compared with the normal group, the spleen values ​​in the induced group increased significantly by about 2 times, and decreased by about 13% and 19% in the PE200 and PE400 administration groups, respectively.

[0085] In this experiment, dexamethasone, a type of adrenocortical steroid drug used as a positive control group, is used as a therapeutic drug for various allergic diseases. However, dexamethasone causes multiple side effects, one of which is increasing the rate of protein breakdown in muscles and inducing skeletal muscle loss. Therefore, in order to confirm whether the side effects of steroid therapeutic agents are also induced in this animal model, the soleus muscle, which is a slow-twitch fiber, and the gastrocnemius muscle, which is a fast-twitch fiber, were removed from the calf muscles to measure muscle mass. Figure 6 As shown, a significant decrease in muscle mass was observed in the DEX-administered group compared to the induced group, but no decrease in muscle mass was observed in the PE-administered group.

[0086] Therefore, it was confirmed that PE can suppress spleen enlargement by inhibiting T lymphocyte activity, and unlike steroid drugs, the risk of side effects can be minimized when used for a long time.

[0087] 10. Blood analysis

[0088] After anesthetizing mice, collect blood from the eye using an orbital blood collection capillary tube or from the inferior vena cava. After standing at room temperature for 1 hour, separate the serum from the blood by centrifugation (1000g, 10 minutes, 4°C). Then, perform the assay according to the ELISA kit protocol and measure the OD value of each well using a spectrophotometer.

[0089] It is well known that overexpression of immunoglobulin E (IgE) produced in B cells is the most representative sign of atopic dermatitis and is proportional to the clinical severity of the patient. Figure 7 As shown, the expression of IgE in the induced group was significantly increased compared to the normal group, and the PE400-administered group showed a significant decrease of approximately 20% compared to the induced group, indicating that PE has the effect of improving atopic dermatitis symptoms.

[0090] 11. Statistical Analysis

[0091] All results are expressed as mean ± standard error, and the significance between the mean values ​​of each group was tested using t-test. When the p value was < 0.05, it was considered statistically significant.

Claims

1. A composition for preventing or treating atopic dermatitis, characterized in that Contains human placenta extract as the active ingredient.

2. A composition for inhibiting the production of TARC or RANTES, characterized in that: Contains human placenta extract as an active ingredient, produced through an allergic immune response.

3. A composition for increasing the production of Treg cytokine TGF-β1, characterized in that: Contains human placenta extract as an active ingredient, which is an immunomodulatory indicator.

4. The composition according to claim 1, characterized in that The above composition inhibits the production of TARC or RANTES produced by an allergic immune response.

5. The composition according to claim 1, characterized in that The above composition increases the production of TGF-β1, a Treg cytokine that is an indicator of immunoregulation.

6. The composition according to claim 1, characterized in that The above composition increases the decreased Th1 cytokine / Th2 cytokine ratio.

7. The composition according to claim 1, characterized in that The above composition inhibits the production of IgE in the blood.

8. The composition according to claim 1, characterized in that The above composition reduces dermatitis scores in atopic dermatitis symptoms.

9. The composition according to claim 1, characterized in that It is a human placenta extract prepared by acetone defatting and then hydrolyzing with pepsin and hydrochloric acid.

10. The composition according to claim 2, characterized in that It is a human placenta extract prepared by acetone defatting and then hydrolyzing with pepsin and hydrochloric acid.

11. The composition according to claim 3, characterized in that It is a human placenta extract prepared by acetone defatting and then hydrolyzing with pepsin and hydrochloric acid.

12. The pharmaceutical composition according to claim 1, characterized in that The dosage form of the above composition is one selected from the group consisting of injection, cream, gel, patch, spray, ointment, plaster, lotion, microneedle, roller and a combination thereof.

13. The pharmaceutical composition according to claim 2, characterized in that The dosage form of the above composition is one selected from the group consisting of injection, cream, gel, patch, spray, ointment, plaster, lotion, microneedle, roller and a combination thereof.

14. The pharmaceutical composition according to claim 3, characterized in that The dosage form of the above composition is one selected from the group consisting of injection, cream, gel, patch, spray, ointment, plaster, lotion, microneedle, roller and a combination thereof.

15. A food composition for improving atopic dermatitis, characterized in that: Contains human placenta extract as an active ingredient, which is prepared by defatting with acetone and then hydrolyzing with pepsin and hydrochloric acid.

16. The food composition according to claim 15, characterized in that The food composition has a dosage form selected from the group consisting of functional food, nutritional supplement, health food, food additive, feed and a combination thereof.

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