Application of pomegranate juice or cold-dried product thereof in preparation of medicine for treating allergic contact dermatitis and medicine for treating allergic contact dermatitis

Abstract

The invention provides application of pomegranate juice or a cold-dried product thereof in preparation of a medicine for treating allergic contact dermatitis and the medicine for treating the allergic contact dermatitis. The pomegranate juice freeze-dried product has the advantages of being natural, safe, free of hormone dependence and the like in the aspect of treating the allergic contact dermatitis, and compared with traditional medicine treatment, the pomegranate juice freeze-dried product is more suitable for long-term use and remarkable in comprehensive curative effect.

Description

Technical Field

[0001] The invention relates to use of pomegranate juice or a freeze-dried product thereof in preparing a medicine for treating allergic contact dermatitis, and a medicine for treating allergic contact dermatitis. Background Art

[0002] Pomegranate (Punica granatum L.) belongs to the genus Punica (Punica L.) of the family Punicaceae. Pomegranate fruit is rich in various nutrients such as protein, amino acids, vitamins, lipids, sugars, crude fiber, etc. In addition, pomegranate is also rich in active substances such as flavonoids, anthocyanins, steroids, stigmasterol, etc., and is a raw material for manufacturing medicines, cosmetics and functional foods. The peel accounts for about 40% of the fresh weight, and is rich in biologically active ingredients such as flavonoids, alkaloids, phenolic substances, etc., and has pharmacological functions such as antibacterial, antioxidant and hypolipidemic. At present, a variety of biologically active and nutritious ingredients such as polyphenol compounds, organic acids, alkaloids, steroid hormones, vitamins and minerals have been found in pomegranates. Its various components vary greatly with different varieties and origins, and the types and contents of chemical components contained in different parts are different. Pomegranate is one of the fruits rich in polyphenols, and the types and contents of polyphenol compounds in pomegranates from different origins are different. The types of polyphenols in pomegranate are mainly tannins, flavonoids, anthocyanidins, etc. Among them, the polyphenol content of pomegranate peel (2.45%-24.9%) is the highest, mainly tannins and flavonoids, and the polyphenol content of pomegranate juice is between 0.2%-1.0%, mainly anthocyanidins; the polyphenol content in pomegranate seed oil is very low, only 0.5mg / g-2.2mg / g, mainly flavonoids and proanthocyanidins. Mao Juanjuan, et al., Clinical observation of 30 cases of atopic dermatitis treated with pomegranate peel ointment, "Hunan Journal of Traditional Chinese Medicine" 2014 No. 4, disclosed the treatment of atopic dermatitis with pomegranate peel ointment.

[0003] Atopic dermatitis (also known as eczema) and allergic contact dermatitis ACD are two different types of skin inflammation. Atopic dermatitis: It is a chronic, recurrent skin disease that is often associated with genetic factors and immune system abnormalities. It is common in children, but can also persist into adulthood. It is characterized by dry skin, itching, redness, swelling and scaling. Allergic contact dermatitis: It is a local inflammatory reaction caused by skin contact with allergens. It usually appears within hours to days after contact with allergens, manifested as redness, itching, blisters, etc. Clinical manifestations: Atopic dermatitis: The skin is usually dry, common in the elbows, knees, face and other parts, accompanied by severe itching, and skin thickening and pigmentation may occur. Allergic contact dermatitis: Symptoms usually appear at the site of contact with allergens, may be accompanied by blisters, exudation and crusting, and the symptoms are relatively limited. In terms of treatment, atopic dermatitis: It is mainly treated with moisturizing, topical steroids, immunomodulators, etc. Allergic contact dermatitis: Avoid contact with the allergen and use topical corticosteroids and antihistamines to relieve symptoms.

[0004] There are currently no reports on the use of pomegranate in the treatment of allergic contact dermatitis. Summary of the invention

[0005] The invention provides a new use of pomegranate.

[0006] The invention provides use of pomegranate juice or a freeze-dried product thereof in preparing a medicine for treating allergic contact dermatitis.

[0007] Wherein, the pomegranate juice is juice squeezed from fresh pomegranate fruits.

[0008] Wherein, the pomegranate fruit comprises pomegranate peel, a separating membrane, pomegranate pulp (aril), and pomegranate seeds.

[0009] Wherein, the preparation method of the freeze-dried product comprises the following steps:

[0010] The pomegranates are taken for juicing, and then the obtained juice is placed in a freezer at -20°C for storage until it is completely frozen; then, the frozen pomegranate juice is freeze-dried in a freeze dryer at -80°C to finally obtain a pomegranate concentrate freeze-dried product.

[0011] Wherein, the preparation of the medicine is an oral preparation.

[0012] The invention also provides a medicine for treating allergic contact dermatitis, which contains pomegranate juice or its lyophilized product.

[0013] Wherein, the pomegranate juice is juice squeezed from fresh pomegranate fruits.

[0014] Wherein, the pomegranate fruit comprises pomegranate peel, a separating membrane, pomegranate pulp (aril), and pomegranate seeds.

[0015] Wherein, the preparation method of the freeze-dried product comprises the following steps:

[0016] The pomegranates are taken for juicing, and then the obtained juice is placed in a freezer at -20°C for storage until it is completely frozen; then, the frozen pomegranate juice is freeze-dried in a freeze dryer at -80°C to finally obtain a pomegranate concentrate freeze-dried product.

[0017] Wherein, the preparation of the medicine is an oral preparation.

[0018] The pomegranate juice freeze-dried product of the present invention has unique advantages in treating allergic contact dermatitis. Compared with other treatment methods, its advantages are mainly reflected in the following aspects:

[0019] 1. Natural ingredients, high safety: Pomegranate juice freeze-dried products and pomegranate polyphenols have natural anti-inflammatory and antioxidant properties, which can effectively reduce skin inflammation and oxidative stress. Compared with traditional chemically synthesized drugs, pomegranate juice freeze-dried products will hardly cause serious side effects during use.

[0020] 2. Anti-inflammatory effect of polyphenols: Pomegranate juice freeze-dried products are rich in polyphenols, such as ellagitannins, flavonoids, etc. These ingredients have significant anti-inflammatory and antioxidant abilities and can effectively inhibit inflammatory reactions. This patent study shows that pomegranate juice freeze-dried products have a good inhibitory effect on allergic contact dermatitis.

[0021] 3. Promote skin repair: The nutrients in freeze-dried pomegranate juice, such as vitamin C and polyphenols, can not only reduce inflammation, but also promote skin repair and regeneration. This is different from traditional medicines that mainly focus on suppressing inflammation. Pomegranate can also help the skin recover during treatment.

[0022] 4. No hormone dependence: Traditional glucocorticoid drugs (such as dexamethasone) may cause hormone-dependent dermatitis, skin atrophy and other side effects after long-term use. As a natural plant ingredient, pomegranate juice freeze-dried products do not have hormone dependence problems and are suitable for long-term use.

[0023] 5. Wide range of applications: Pomegranate juice freeze-dried products are not only suitable for mild and moderate allergic contact dermatitis, but can also be used to prevent and assist in the treatment of allergic contact dermatitis.

[0024] 6. No obvious contraindications: Pomegranate juice freeze-dried products have no obvious contraindications during use and are suitable for most people, including pregnant women and children.

[0025] In summary, freeze-dried pomegranate juice has the advantages of being natural, safe, and non-hormone-dependent in the treatment of allergic contact dermatitis. Compared with traditional drug treatment, it is more suitable for long-term use and has significant overall therapeutic effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 .. Flow chart of the DNFB-induced ACD mouse model experiment;

[0027] Figure 2 . Changes in mouse body weight;

[0028] Figure 3 . Mouse spleen index;

[0029] Figure 4 .Statistical analysis of mouse scratching behavior on day 14;

[0030] Figure 5 . Skin pictures of mice at the modeling site on day 14 (A) and statistical analysis of the average skin injury scores of mice from day 0 to day 14 (B);

[0031] Figure 6 . Picture of mouse ear tissue on day 14 (A), statistical analysis of thickness difference between both ears of mice (B), and statistical analysis of swelling degree of diseased ears of mice (C);

[0032] Figure 7 . Pathological observation of mouse lesion ear skin by H&E staining (A, B), pathological observation of mouse lesion ear skin by TB staining (C), statistical analysis of mouse lesion ear epidermal thickness (D), and statistical analysis of mouse lesion ear mast cell count (D);

[0033] Figure 8 .The expression levels of serum immunoglobulin Ig-E (A), serum histamine (B), IL-4 (C), IL-13 (D), TNF-α (E), and IL-1β (F) in the affected ear tissue of mice. DETAILED DESCRIPTION

[0034] Example 1 Preparation of freeze-dried pomegranate juice of the present invention

[0035] Pomegranates produced in Huili, Sichuan Province were selected for juicing, and then the obtained juice was frozen and stored at -20°C until it was completely frozen. Subsequently, the frozen pomegranate juice was freeze-dried in a freeze dryer at -80°C to finally obtain pomegranate concentrate.

[0036] The beneficial effects of the present invention are demonstrated by the following efficacy test.

[0037] Test Example 1 Application of the freeze-dried pomegranate juice of the present invention to allergic contact dermatitis (ACD)

[0038] BALB / c female mice (SPF grade) aged 6-8 weeks, weighing 19.00±2.00 g, were purchased from Spelford Biotechnology Co., Ltd. (Beijing, China). They were used for the experiment after one week of adaptive feeding. The mice were kept under specific conditions throughout the whole process, with a temperature of 25±2°C, a relative humidity of 50±10%, a light-dark cycle of 12 h, and free access to standard rodent chow and water. All procedures followed international ethical guidelines and were approved by the Ethics Committee of Chengdu University.

[0039] 1. The ACD model is established as follows:

[0040] Four days before the experiment, the hair on the right dorsal abdomen of the mice was shaved in an area of ​​1.5 cm × 2 cm. The mouse skin was pre-sensitized and sensitized with 2,4-dinitrofluorobenzene (DNFB).

[0041] After shaving, the mice were randomly divided into 5 groups after adaptive feeding for 3 days, namely, blank group (CON), modeling group (DNFB), positive drug dexamethasone group (DEX), low-dose group of pomegranate concentrate prepared in Example 1 (PJL, 100 mg / kg) and high-dose group of pomegranate concentrate (PJH, 200 mg / kg). Except for the blank group (CON), the other 4 groups of mice were evenly smeared with 50 μL of 0.5% DNFB solution (dissolved in acetone: olive oil, 4:1, v / v) on the smooth skin after depilation for pre-sensitization treatment. The blank group mice were only smeared with the same volume of acetone: olive oil solvent (4:1, v / v) as a control. The operation was repeated twice and the treatment was stopped for 4 days. Except for the blank group, all mice showed hypersensitivity reactions of skin allergy, redness, severe itching and scratching.

[0042] Starting from the 6th day of sensitization, the right dorsal and abdominal modeling site and the inner side of the right auricle (lesioned ear) received 20 μL of 0.2% DNFB solution (dissolved in acetone: olive oil solvent, 4:1, v / v). The mice in the blank group were treated with an equal volume of acetone / olive oil (4:1, v / v) solvent alone. The sensitization process was repeated every 2 days until the 14th day of the experiment. During the sensitization phase from the 6th day to the 14th day, the mice in the CON group and the DNFB group received 0.2 mL of saline solution by gavage every day; the mice in the DEX group received 0.2 mL of 5 mg / kg dexamethasone saline solution by intramuscular injection every day; the mice in the PJL group and the PJG group received 0.2 mL of low-dose (100 mg / kg) and high-dose (200 mg / kg) concentrated pomegranate juice by gavage, respectively (see Figure 1 ).

[0043] The weight of mice was recorded every day during the experiment. After fasting for 12 hours, the mice were sacrificed under anesthesia after blood was collected from their orbits. The mice were immediately placed on ice and their ears were cut off along the base of the ear tissue with medical scissors. The ear tissues and spleen of each mouse were collected, weighed, and stored at -80°C for later use.

[0044] 2. Experimental results

[0045] 2.1. Effect of the freeze-dried pomegranate juice of the present invention on the body weight of mice with ACD disease

[0046] The mouse body weight results were as follows Figure 2 As shown in the figure, at the starting point of the experiment (day 0), the weight levels of mice in each group were basically the same, with no significant difference. The weight of mice in the CON group continued to increase throughout the modeling period of 0-14 days. This result shows that the solvent applied to the CON group (acetone: olive oil, 4:1, v / v) has no significant effect on the growth and weight changes of mice.

[0047] On the 3rd day, the weight of the other four groups of mice, except the CON group, decreased significantly after modeling with 0.5% DNFB solution, and the downward trend remained consistent. On the 6th day, after two DNFB modeling treatments, the modeling process was stopped, and the mice were not treated for 4 days. At this time, the weight of the four groups of mice rebounded significantly, and the weight levels between the groups remained roughly the same. This phenomenon shows that DNFB modeling causes mice to lose weight, but after stopping the modeling, the mice can gradually recover their weight during this period.

[0048] Starting from the 6th day, except for the CON group, the other four groups of mice received 0.2% DNFB sensitization treatment every two days. During this period, the mice in the DNFB group were gavaged with normal saline, and their body weight showed a linear downward trend, and the initial decline was extremely rapid, and the subsequent decline rate gradually slowed down. Specifically, the average body weight of mice in the DNFB group decreased by 3.44±0.27g within 6-14 days. In contrast, the weight of mice in the DEX group only decreased slightly after receiving dexamethasone injection. Within 6-14 days, the average body weight of mice in the DEX group only decreased by 0.98±0.80g. This shows that dexamethasone can alleviate the weight loss induced by DNFB to a certain extent and basically maintain the weight level of mice.

[0049] During the observation period of 6-14 days, the body weight of mice in the PJL group and the PJH group showed a trend of first decreasing and then increasing. Specifically, the weight loss trend of the PJH group from 6 to 12 days was significantly gentler than that of the PJL group; and after the 12th day, the weight gain rate of the PJH group was also significantly faster than that of the PJL group. This result shows that the PJH group is superior to the PJL group in terms of body weight. It is particularly noteworthy that on the 14th day, the body weight of mice in the PJH group had exceeded that of the DEX group and was higher than that of the PJL group. This shows that when the dose of pomegranate concentrate freeze-dried product is high enough, the body weight of mice can exceed that of the DEX group, which may enable the body to recover to a certain extent, thereby effectively alleviating the discomfort caused by DNFB to the mice.

[0050] 2.2. Effect of the freeze-dried pomegranate juice of the present invention on the spleen index of ACD disease mice

[0051] The mouse spleen was weighed and the spleen organ index was calculated using the following formula:

[0052]

[0053] like Figure 3 As shown in the figure, compared with the CON group, the spleen index of mice in the DNFB group increased significantly (P<0.01). Under the induction of DNFB, the spleen will swell, resulting in an increase in spleen weight, which leads to an increase in spleen index. Compared with the DNFB group, the spleen index level in the DEX group decreased significantly (P<0.01). Compared with the DNFB group, both the PJL group and the PJH group could significantly reduce the swelling of the spleen (P<0.01), among which the effect of the PJH group was particularly prominent. The above results show that pomegranate concentrated freeze-dried product can significantly reduce the edema of the spleen and further reduce the spleen index, which has a certain inhibitory effect on the DNFB-induced spleen swelling.

[0054] 2.3. Effect of the freeze-dried pomegranate juice of the present invention on the scratching behavior of ACD disease mice

[0055] In order to explore the effect of freeze-dried pomegranate juice on the scratching behavior of ACD mice, mice were placed in a transparent observation cage for adaptation 30 minutes before sensitization on the 14th day of modeling. Then, DNFB modeling drugs or vehicle solvents (acetone: olive oil, 4:1, v / v) were dripped and smeared on the right dorsal and abdominal modeling sites and the inner side of the right auricle. After confirming that the mice were in good physiological condition, they were immediately placed in a box and recorded with a camera for 1 hour in a quiet environment. The scratching behavior of the mice was observed and the number of scratches in 1 hour was counted. The mouse lifted its right hind foot to scratch the abdomen or neck and put it down, which was counted as 1 time for statistical processing.

[0056] Immediately after induction, the scratching behavior of mice was recorded using a video camera, e.g. Figure 4The scratching frequency of mice in the DNFB group showed significant differences compared with the CON group, and the scratching frequency increased significantly (P<0.01). At the same time, the DEX group, 100 and 200 mg / kg doses of PJL and PJH groups can alleviate the spontaneous scratching behavior of ACD mice, and the inhibitory effect of pomegranate concentrated freeze-dried product on the scratching frequency of ACD mice was significantly different (P<0.01), showing a dose-dependent manner. The scratching frequency of the PJL and PJH groups was 123±5 and 83±9 times, respectively, and the high-dose PJH group had the most significant effect.

[0057] 2.4. Effect of the freeze-dried pomegranate juice of the present invention on the skin lesion score of ACD disease mice

[0058] The skin of the right dorsal and abdominal areas of the mice was scored according to the severity of erythema, scratches, bleeding and blood scabs, which were scored as 0 (none), 1 (mild), 2 (moderate) and 3 (severe). The sum of the above four scores was calculated, and the scores of each day were added up to a total score, which was the skin injury score.

[0059] The skin condition of mice on day 14 is as follows Figure 5 There was no obvious skin damage in the CON group mice; on the contrary, the DNFB group mice showed obvious ACD symptoms, with erythema, ulceration, and scabs on the skin of the modeling site. The ulcers were more severe with scabs, hardening, and epidermal shedding. In addition, the skin injury score of the DNFB group mice was significantly increased, which was significantly different from that of the CON group (P<0.01), indicating that the DNFB group had more severe skin damage.

[0060] The skin injury scores of mice in the DEX group were significantly lower than those in the DNFB group (P<0.01), and the damage was significantly alleviated. The scratches and erythema areas of mice in the DEX group were smaller, and only a small part of the skin was bleeding, ulcerated, and scabbed. This phenomenon shows that dexamethasone has a significant positive effect on DNFB-induced dermatitis damage, can effectively reduce skin inflammation and damage, and accelerate the skin recovery process.

[0061] The skin injury scores of the PJL and PJH groups were significantly lower than those of the DNFB group (P<0.01), indicating that pomegranate concentrate freeze-dried products have a significant inhibitory effect on skin injury. In addition, the skin injury of the PJ high-dose group (PJH) was better than that of the low-dose group (PJL), as shown by more complete scab shedding and smaller wounds. The skin injury score also showed that the PJH group was significantly lower than the PJL group. These results show that pomegranate concentrate freeze-dried products can effectively treat DNFB-induced allergic contact dermatitis (ACD) skin injuries, and the therapeutic effect is positively correlated with the dose.

[0062] 2.5. Effect of the freeze-dried pomegranate juice of the present invention on the thickness and weight of ear tissue in mice with ACD disease

[0063] On the 14th day of the experiment, 2 hours after the allergen was applied, the thickness of the left and right ear tissues of the mice was measured using a digital micrometer with a sensitivity of 0.001 mm. Each measurement was made at the same position (3 mm from the middle of both ears to the ear edge), perpendicular to the cartilage, and repeated three times to ensure the accuracy of the data. The formula is as follows:

[0064] Thickness difference between both ears = thickness of the right ear of the diseased ear (mm) - thickness of the left ear of the normal ear (mm)

[0065]

[0066] Compared with the CON group, the difference in thickness between the left and right ears of mice in the DNFB group increased significantly (P<0.01), and the degree of swelling of the affected ear also increased significantly (P<0.01). Specifically, the thickness of the affected right ear in the DNFB group was about 208±16μm higher than that of the left ear on average, while the difference in thickness between the left and right ears in the control group was only 10μm, which was almost negligible. This result indicates that the solvent used in the experimental CON group (acetone: olive oil, 4:1, v / v) has no significant effect on ear tissue, further confirming the successful establishment of the ear swelling effect in the DNFB-induced ACD model.

[0067] Compared with the DNFB group, the difference in bilateral ear thickness of mice in the DEX group was significantly reduced (P<0.01), the swelling of the diseased ears was also significantly reduced (P<0.01), and the difference in bilateral ear thickness and weight was narrowed.

[0068] In addition, from Figure 6 It can be observed that the PJL group and the PJH group can effectively reduce the swelling of the diseased ear, effectively reduce the thickness and weight of the diseased ear, and thus significantly reduce the difference in thickness between the two ears (P<0.01), and the swelling of the diseased ear is also significantly reduced (P<0.01). Further analysis found that the difference in thickness between the two ears in the low-dose PJ group (PJL group) and the high-dose PJ group (PJH group) was 140±51μm and 104±28μm, respectively. This result shows that the higher the dose of pomegranate concentrated freeze-dried product, the smaller the difference in thickness between the two ears, and the milder the swelling of the ear tissue, showing a significant dose-related effect.

[0069] 2.6. Observation on the effect of freeze-dried pomegranate juice on ear pathology in mice with ACD

[0070] The middle part of the diseased ear tissue of each group of mice was taken, with a width of about 1.5 mm, and fixed with 4% paraformaldehyde. After fixation, it was dehydrated and embedded into wax blocks, cut into 5 μm slices by a slicer, and then stained with hematoxylin-eosin (HE) and toluidine blue (TB), respectively. The slices were observed under an optical microscope and the scanned images were recorded. ImageJ was used to measure the epidermal thickness in H&E stained slices and statistically analyzed. Iviewer was used to analyze the number of mast cells in toluidine blue stained slices and statistically analyzed.

[0071] H&E staining results showed that the structure of each layer of the ear tissue skin of the CON group mice was clear, with no obvious thickening, clear edges, and no obvious pathological changes. The ear tissue lesions of the DNFB group mice showed more severe congestion and swelling compared with the CON group, unclear boundaries between the epidermis and dermis, hyperkeratosis of the skin epidermis, lichenoid infiltration, obvious thickening of the epidermis (P<0.01), and obvious inflammatory infiltration. There were a large number of monocytes and lymphocytes infiltrating between the epidermis and dermis, indicating that DNFB had significant damage to the mouse skin. After dexamethasone treatment, the pathological damage in the DEX group was significantly improved compared with the DNFB group, and the thickness of the epidermis was significantly reduced (P<0.01). After oral feeding with PJ, the pathological damage in the PJL group and the PJH group was reduced compared with the DNFB group, and the results showed that the thickness of the epidermis of the skin was significantly reduced (P<0.01). In particular, the boundaries of the skin layers of the PJH group were clearer, the infiltration of inflammatory cells was reduced, and no severe congestion and swelling occurred.

[0072] Mast cell staining results showed that the number of mast cell infiltration in the dermis of the ear tissue in the DNFB group was significantly higher than that in the CON group (P<0.01). The mast cells were densely distributed, round or oval, with small nuclei, and the cytoplasm was filled with blue-purple particles evenly distributed around the nucleus. The epidermis and dermis of the skin of the mice in the DEX group did not show obvious thickening compared with the DNFB group, and the number of mast cell infiltration was significantly reduced (P<0.01). Compared with the DNFB group, the PJL group and the PJH group could significantly improve the mast cell infiltration in the skin of ACD mice (P<0.01).

[0073] The results of H&E staining and mast cell staining showed that compared with the DNFB group, the treatment with pomegranate concentrated freeze-dried products not only reduced the increased epidermal thickness and inflammatory cell infiltration caused by skin damage in ACD mice, but also effectively improved mast cell infiltration. Finally, the skin damage of mouse ear tissue was significantly reduced, and the recovery effect of the high-dose PJH group was more obvious. This result suggests that pomegranate freeze-dried products can effectively improve the skin pathological state of contact allergic dermatitis induced by DNFB, and its improvement effect is dose-related (see Figure 7 ).

[0074] 2.7. Effects of the freeze-dried pomegranate juice of the present invention on immunoglobulins and cytokines in mice with ACD disease

[0075] After the whole blood sample was allowed to stand at room temperature for 1 hour, it was centrifuged at 1000× for 20 minutes to obtain the upper serum. Immunoglobulin (IgE) and histamine levels were determined according to the instructions of the enzyme-linked immunosorbent assay (ELISA) kit.

[0076] The remaining mouse lesion ear tissues were taken and added with pre-cooled PBS (0.01M, pH=7.4) at a weight (g): volume (mL) ratio of 1:9, and a 10% tissue homogenate was prepared using a freezing grinder, and the supernatant was obtained by centrifugation. According to the instructions of the enzyme-linked immunosorbent assay (ELISA) kit, the levels of cytokines interleukin-4 (IL-4), IL-13, tumor necrosis factor-alpha (TNF-α), and interleukin-1beta (IL-1β) in the ear tissues of mice in each group were determined.

[0077] The results are as follows Figure 8 As shown in the figure, compared with the CON group, the levels of immunoglobulin IgE and histamine in the serum of mice in the DNFB group and the levels of cytokines IL-4, IL-13, TNF-α and IL-1β in the lesion ear tissue were significantly increased (P<0.01), indicating that DNFB successfully established the ACD disease model in mice. Compared with the DNFB group, the levels of immunoglobulin IgE and histamine in the serum of mice and the levels of cytokines IL-4, IL-13, TNF-α and IL-1β in the lesion ear tissue were significantly decreased in the DEX group, PJL group and PJH group (P<0.01).

[0078] The present invention further utilizes the DNFB-induced allergic contact dermatitis mouse model to explore the therapeutic effects of the pomegranate juice freeze-dried product PJL group (low-dose group 100 mg / kg) and PJH group (high-dose group 200 mg / kg) on ​​allergic contact dermatitis. The results show that compared with the DNFB group modeling mice, the weight of the mice can be increased; the spleen index of the mice can be reduced to avoid excessive swelling of the spleen; the itching can be reduced, thereby reducing the scratching behavior of the mice, avoiding further damage to the skin, and reducing the skin damage score; the thickness and weight of the mouse diseased ear tissue can be reduced, and the swelling of the diseased ear can be reduced. At the same time, the ear tissue pathology results also prove that the pomegranate juice freeze-dried product can reduce the epidermal thickness and inflammatory cell infiltration of the diseased ear tissue due to skin damage, and can also effectively improve the mast cell infiltration phenomenon. Freeze-dried pomegranate juice can significantly reduce the levels of immunoglobulin IgE and histamine in mouse serum and cytokines IL-4, IL-13, TNF-α and IL-1β in diseased ear tissue. This patent shows that freeze-dried pomegranate juice has a significant inhibitory effect on allergic contact dermatitis (ACD).

Claims

1. Use of pomegranate juice or its freeze-dried product in preparing medicine for treating allergic contact dermatitis.

2. The use according to claim 1, characterized in that: The pomegranate juice is juice squeezed from fresh pomegranate fruits.

3. The use according to claim 2, characterized in that: The pomegranate fruit comprises pomegranate peel, a separation membrane, pomegranate pulp (aril) and pomegranate seeds.

4. The use according to claim 1, characterized in that: The preparation method of the freeze-dried product comprises the following steps: The pomegranates are taken for juicing, and then the obtained juice is placed in a freezer at -20°C for storage until it is completely frozen; then, the frozen pomegranate juice is freeze-dried in a freeze dryer at -80°C to finally obtain a pomegranate concentrate freeze-dried product.

5. The use according to any one of claims 1 to 4, characterized in that: The preparation of the medicine is an oral preparation.

6. A drug for treating allergic contact dermatitis, characterized in that: It contains pomegranate juice or its frozen dried form.

7. The drug for treating allergic contact dermatitis according to claim 6, characterized in that: The pomegranate juice is juice squeezed from fresh pomegranate fruits.

8. The drug for treating allergic contact dermatitis according to claim 7, characterized in that: The pomegranate fruit comprises pomegranate peel, a separation membrane, pomegranate pulp (aril) and pomegranate seeds.

9. The drug for treating allergic contact dermatitis according to claim 6, characterized in that: The preparation method of the freeze-dried product comprises the following steps: The pomegranates are taken for juicing, and then the obtained juice is placed in a freezer at -20°C for storage until it is completely frozen; then, the frozen pomegranate juice is freeze-dried in a freeze dryer at -80°C to finally obtain a pomegranate concentrate freeze-dried product.

10. The drug for treating allergic contact dermatitis according to any one of claims 6 to 9, characterized in that: The preparation of the medicine is an oral preparation.

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