[논문]하수오 추출물(Polygonum multiflorum)이 아토피 피부염 유발 NC/Nga 생쥐의 알레르기 반응에 미치는 영향

문지선; 최은영

doi:10.12925/jkocs.2020.37.1.66

하수오 추출물(Polygonum multiflorum)이 아토피 피부염 유발 NC/Nga 생쥐의 알레르기 반응에 미치는 영향
Effect of Polygonum multipolarum extract on the allergic reaction of NC/Nga mice causing atopic dermatitis 원문보기

Journal of the Korean Applied Science and Technology = 한국응용과학기술학회지, v.37 no.1, 2020년, pp.66 - 75

문지선 (중원대학교 의료뷰티케어학과) , 최은영 (알엑스에이피코리아)

Abstract ▼ AI-Helper

Atopic dermatitis (AD) usually develops in patients with an individual or family history of allergic diseases, and is characterized by chronic relapsing inflammation seen specially in childhood, association with IgE hyperproduction and precipitation by environmental factors. and wished to examine closely effect that Polygonum multiflorum isolated PM-E and PM-70M orally adminstration used to atopy dermatitis disease patient get in atopy eruption control experimentally. Atopic dermatitis is a chronically relapsing inflammatory skin disease. Animal models induced by relevant allergens play a very important role in the elucidation of the disease. This study was investigated the anti-allergic effect of PM-E and PM-70M on BMAC induced atopic dermatitis in NC/Nga mice. We summerized as the follow. PM-E and PM-70M significantly reduced the skin number of total cell number, CD4+ and CD11b+/Gr-1 cell compared with positive control and decreased the invasion of CD4+ cell in dorsal skin tissue compared with positive control group by using immunohistochemical staining and chemokine such as eotaxin and CCR3 compared with positive control group. PM-E and PM-70M markedly suppressed invasion and edema of leukocytes and mast cell in dorsal skin. Taken together, these findings suggested that PM-E and PM-70M has an anti-allergic activity and this might be useful for the clinical application to treat allergic diseases such as atopic dermatitis.

주제어

표/그림 (6)

그림 Fig. 1 Experimental Design for the Induction of Atopic Dermatitis like NC/Nga Mouse.
표 Table 1. Primer Sequence
그림 Fig. 2. Total cell number of dorsal skin in atopic dermatitis skin lesions in NC/Nga mouse.
그림 Fig. 3. Effects of PM-E & PM-70% MFL treatment on changes of absolute numbers in dorsal skin cells in NC/Nga mouse(A). Immunohistochemical staining of the skin stained with CD4⁺ Th cells of dorsal skin in atopic dermatitis skin lesions in NC/Nga mouse(B).
그림 Fig. 4. WBC differential counting in atopic dermatitis skin lesions in NC/Nga mouse.
그림 Fig. 5. Effects of PM-E & PM-70%MFL treatment on Eotaxin2, and CCR3, mRNA expression in NC/Nga mouse.

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가설 설정

1. The PM-70% MFL administration group showed a decrease in skin total cell compared to the control group.

제안 방법

Total cell number of dorsal skin in atopic dermatitis skin lesions in NC/Nga mouse. Atopic dermatitis NC/Nga mouse was induced by BMAC treatment in the dorsal skin, non treated BMAC (SPF-normal, Nr), BMAC treatment for 3 weeks (control), BMAC treatment for 3 weeks with CsA (10 mg/kg, i.p), BMAC treatment for 3 weeks with PM-E (200 mg/kg) orally administration, and BMAC treatment for 3 weeks with PM-70%MFL (75 mg/kg) orally administration for 3 weeks.
BMAC was injected with 4% SDS solution twice a week for a total of three weeks (10 to 13 weeks of age), and 200 μl each was applied after 2 hours.
After that, the mice were anesthetized with chloral hydrate (10%), anesthetized, and the dorsal neck area was depilated cleanly. Biostir Mite antigen cream (hereinafter referred to as BMAC: Dermatophagoides farinae crude extract) provided by the central laboratory animal was evenly applied to the back and neck area twice a week for 3 weeks (10 to 13 weeks of age), and 4% SDS 2 hours before application. The mice were used as a control by destroying the skin layer to induce dermatitis by spraying the solution (Fig.
BMAC was injected with 4% SDS solution twice a week for a total of three weeks (10 to 13 weeks of age), and 200 μl each was applied after 2 hours. Dermatitis rash was confirmed 2 weeks after the start of BMAC application (12 weeks of age), and once every 3 weeks from 12 weeks to 15 weeks of age, CsA (10 mg / kg), PM-E (200 mg / kg) and PM, respectively. -70% MFL (75 mg / kg) was administered.
Using the Hasuo sample (PM-E, PM) as a sample, the author investigated the effects of changes in immune cell number, chemokine expression, and inflammation of skin tissues using atopic dermatitis animal model induced by BMAC, and came to the same conclusion.

대상 데이터

Reagents used in this experiment were diethyl pyrocarbonate (DEPC), chloroform, trichloroacetic acid, isopropanol, Tris-HCl, KCl, MgCl2, ACK lysis solution, DMEM culture, dulbecco's phosphate buffered saline (D-PBS), Sulforhodamin B (SRB), 2-isopropanol, Sodium dodecyl sulfate (SDS), PMA, Ionomycin, FK506, and antibiotics were manufactured by Sigma (USA).
The equipment used in this experiment was a boiling water extractor (Daewoong, Korea), rotary vaccum evaporator (Büchi B-480, Switzerland), freeze dryer (EYELA FDU-540, Japan), CO2 incubator (Forma scientific Co., USA), clean bench (Vision scientific Co., Korea), autoclave (Sanyo, Japan), micro-pipet (Gilson, France), water bath (Vision scientific Co., Korea), vortex mixer (Vision scientific Co., Korea), spectrophotometer (Shimazue, Japan), centrifuge (Sigma, USA), deep-freezer (Sanyo, Japan), Quantitative Real-Time RT-PCR (Applied Biosystems, USA), ice-maker (Vision scientific Co., Korea), homogenizer (OMNI, USA), plate shaker (Lab-Line, USA), Vario MACS (Bergisch Gladbach, Germany), FACS calibur (BD, USA) and ELISA leader (Molecular Devices, USA).
The experiment was conducted in the NC/Nga-Nr group (Nr) and BMAC-coated control group in which 7-week-old NC/Nga mice were kept in SPF conditions up to 15 weeks of age. BMAC was applied and divided into experimental groups administered PM-E and PM-70% MFL.

데이터처리

Results obtained from the experiments were recorded as mean ± standard error, and significance test was performed using Student's t-test analysis method[9].

이론/모형

After the final clinical visual evaluation, the blood was collected and Biotoxtec measured the number of neutrophils, eosinophils, basophils and lymphocytes, and monocytes. The measurement was performed with an automatic hemocytometer (MS9-5, MELET SCHLOESING, France) using Minos-ST according to the Fonio method[7].

성능/효과

2. The effect of PM-70% MFL on the immune cells through fluorescence flow cytometry in the back skin tissue showed that the number of CD4+, CD11b+/GR-r1+ cells in the skin was significantly reduced compared to the control group.
3. In the PM-70% MFL group, the cell number was significantly decreased due to the decrease of neutrophil, eosinophils, monocytes and basophils even in the effect of leukocyte.
As a result of measuring the absolute cell number of CD4+ cells in the dorsal skin tissue, the control group increased significantly compared to NC/Nga-Nr group, and the CsA-administered group significantly decreased more than 6 times compared to the control group (p<0.001).
As a result, PM-70% MFL showed significant clinical symptom improvement effect through skin inflammation inhibitory effect in BMAC-induced atopic dermatitis animal model, and it was recognized as efficacy for the treatment of atopic dermatitis.
At the end of the experiment (15 weeks), WBC neutrophil and eosinophils ratios were significantly higher in the control group than in the NC/Nga-Nr group. The CsA group was significantly decreased (p<0.
The CsA group was significantly decreased (p<0.01, p<0.001) compared to the NC/Nga-Nr group compared to the control group, and the neutrophil and eosinophils ratios in the PM-E group and the PM-70% MFL group were decreased compared to the control group.

후속연구

Th2-cell responses were obtained by administering Rumex japonicus Houtt herb containing Emodin, chrysophanol, and physcion to NC/Nga mice, atopy-induced mice and skin rashes were reported to be suppressed[5]. Based on these basic data, the author would like to continue to study on the atopic dermatitis.

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