[논문]갈근 에탄올 추출물의 피부 보습 효과에 대한 연구

홍철희; 이성진

doi:10.6114/jkood.2017.30.1.106

갈근 에탄올 추출물의 피부 보습 효과에 대한 연구
Study on the Moisturizing Effects of Puerariae Radix Ethanol Extract 원문보기

韓方眼耳鼻咽喉皮膚科學會誌 = The journal of Korean Medicine Ophthalmology & Otolaryngology & Dermatology, v.30 no.1, 2017년, pp.106 - 117

홍철희 (상지대학교 한의과대학 한방안이비인후피부과학교실) , 이성진 (상지대학교 한의과대학 한방안이비인후피부과학교실)

초록
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Objectives : 갈근의 피부 보습 효과에 대해 알아보기 위해 본 연구를 진행하였다. Methods : HaCaT 세포주에 갈근 에탄올 추출물을 처리하였고 Retinoic Acid $1{\mu}M$을 양성 대조군으로 사용하였다. MTT assay, RT-PCR, Hyaluronidase enzyme assay, HA-ELISA kit를 통해 Hyaluronic Acid의 합성량, Hyaluronidase 저해율, 농도별 Hyaluronic Acid 생성량을 측정하였다. 더불어 실제로 피부에 도포 시 보습 능력이 있는지 등에 대하여 확인하였다. Results : 갈근 에탄올 추출물은 뚜렷한 세포독성을 나타내지는 않았고, 모든 농도에서 Hyaluronic Acid 합성과 관련된 hyaluronic acid synthase 2 유전자가 발현되었고, 실제로 Hyaluronic Acid가 생성되는 것을 확인하였다. 임상 효능 평가에서는 유의성 있는 결과를 얻을 수는 없었다. Conclusions : 갈근을 한방 외용제로 사용할 때 유의성 있는 효과를 나타낼 수 있을 것으로 기대할 수 있으며, 실험 결과를 토대로 갈근의 최적 사용 농도를 결정하기 위한 추가적 연구와 복합 처방으로 사용했을 경우에 대한 연구가 추가적으로 진행된다면 한방 외용제로서 잠재적인 가치가 있을 것으로 사료된다.

주제어

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문제 정의

Although many studies have reported the antioxidant properties^37,38) of Puerariae radix, few studies have focused on its moisturizing effects. Therefore, the purpose of this study was to evaluate the efficacy of Puerariae radix as a moisturizing agent.

가설 설정

1. In the MTT assay, no apparent cytotoxicity was observed.
3. In the hyaluronidase enzyme assay, PREE inhibited HYAL activity.

제안 방법

A pilot study was performed to measure the skin moisturizing effect of Puerariae radix on human skin. Approval for the experiment was obtained from the Institutional Review Board (IRB No.
HAS2 gene expression for the synthesis of HA was determined via Reverse transcription polymerase chain reaction (RT-PCR) while HYAL inhibition rate was determined via hyaluronidase enzyme assay and the amount of HA was measured via an HA ELISA kit. In addition, a pilot study was performed to confirm the moisturizing effects of PREE when applied on normal human skin. If PREE was found to be non-cytotoxic and the amount of HA produced significant, Puerariae radix could be a good raw material for external herbal applications.
In order to evaluate the moisturizing effect of Puerariae radix, HA production was measured and a pilot study were performed. HA content is regulated by continuous synthesis by hyaluronic acid synthase 2 (HAS2) and degradation by hyaluronidase (HYAL)¹⁷⁾.
In this study, to evaluate the moisturizing effect of Puerariae radix, a 70% (v/v) ethanol solution of the same was prepared, and MTT assay, RT-PCR, hyaluronidase enzyme assay, HA ELISA and a pilot study were conducted.
The HA ELISA kit (Echelon, USA) was used according to the manufacturer's protocol. The cells were treated with 50, 100, and 200 ㎍/㎖ PREE for 24 hours. After 30 minutes using a HA ELISA kit, the amount of HA produced was measured in each sample.

대상 데이터

HaCaT cell lines used in this study were purchased from American Type Culture Collection (ATCC, VA, USA). HaCaT cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco BRL, NY, USA) containing 10% fetal bovine serum (FBS, Gibco BRL, NY, USA) and 1% penicillin-streptomycin (PS, Gibco BRL, NY, USA) in 5% CO₂ atmosphere at 37℃.
Puerariae radix was purchased from Pure Mind Co. (Yongchoen, Korea) through Korean Medicine Hospital, Semyeong University and samples were stored at the Joint Laboratory, School of Pharmaceutical Science and Engineering, Semyeong University. For the experiment, 100 g Puerariae radix was immersed in 70% (v / v) ethanol solution and repeated three times during 72 hours.
SMCTC-48-16-01) of the Chemical Trial Center for Bio-industry at Semyeong University. The study was conducted from September 22 to september 23, 2016. The subjects were 12 adult women with normal skin aged 20 to 55 years.
The study was conducted on 12 normal women subjects. Before and 10 minutes after application, the application site and the non-application site were measured three times each and the values averaged.
The study was conducted from September 22 to september 23, 2016. The subjects were 12 adult women with normal skin aged 20 to 55 years. The skin moisture of the forearm was measured.

데이터처리

Statistical analysis was performed using the ANOVA and t-test. Significance was set as p-value < 0.

이론/모형

Hyaluronidase enzyme assay was performed to measure HYAL inhibition rate, using the Morgan-Elson method²¹⁾. In brief, 50 ㎕ of bovine hyaluronidase (8 ㎎/㎖, 0.

성능/효과

2. By RT-PCR, HAS2 gene expression was confirmed at all concentrations.
4. Evaluation of HA synthesis by HA ELISA kit showed that 50 ㎍/㎖ treatment group was the highest concentration and its average value was 1165.54 ㎍/㎖.

후속연구

As the concentration increases, the expression of HA decreases. Therefore further studies are needed to determine the optimal concentration.
There was almost no difference between before application and 10 minutes after application at either site. Thus, further studies with detailed experimental design would be required.

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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