사삼(Adenophorae Radix) 추출물의 발모효과 연구

Abstract

본 연구는 천연물로서 사삼추출물의 발모효과를 알아보고 탈모예방화장품으로서의 개발가능성에 대한 기초자료를 제공하고자 시도되었으며 이를 위해 마우스모델을 통한 발모효능실험과 인체피부도포 임상실험을 진행하였다. 마우스모델을 통한 발모효능실험은 수컷 6주령 C57BL/6 마우스를 실험동물로 이용하였으며, 사삼시료추출은 70%에탄올용액에 침지한 후 그 여과액을 농축, 동결건조시켜 얻은 분말을 사용하였다. 실험물질은 AR-1(사삼추출물 1%), AR-2(사삼추출물 5%), 에탄올, PBS를 이용하였다. 각각의 시료를 실험동물의 탈모부위에 도포하여 체중변화, 피부의 육안적 관찰, 모낭의 현미경적 관찰, 멜라닌색소 침착 정도, 모발두께와 길이 변화, 모발생성과 분포 등을 관찰하였다. 인체피부도포임상실험은 20대-40대의 남성 탈모자를 대상으로 하였으며 동물실험결과 대조군에 비해 발모효과가 유의적으로 높았던 AR-2(사삼추출물 5%)가 함유된 사삼헤어토닉을 제조하여 5주간 탈모부위에 도포하게 하였다. 연구방법으로는 실험전?후 자가기입형태의 설문지와 사삼헤어토닉 품평서를 작성하게 하였으며 이를 통해 사삼헤어토닉 사용전과 후의 모발과 두피의 변화를 비교하였다. 설문문항은 일반적인 특성, 생활습관 및 건강상태 , 스트레스정도, 영양섭식관리, 모발상태 및 모발두피관리습관 , 현재 두피 및 모발의 상태에 관련된 문항으로 구성하였으며 사삼헤어토닉품평서는 사용의 용이성, 피부자극성, 청량감, 세정성, 방향의 선호도, 구입의사여부 등 6개 항목 문항으로 구성하여 응답케 하였다. 1. 마우스모델을 통한 발모효능실험결과는 다음과 같다. 실험마우스의 체중변화(g)는 AR-1(사삼추출물 1%)은 23.52(±0.48), AR-2(사삼추출물 5%)는 24.12(±0.74), ET(에탄올)는 24.00(±0.45), PBS는 23.52(±0.54)로 4가지 사삼추출물용액 시료 간 마우스의 체중은 차이가 없었으며 체중증가량에도 유의한 차이가 없었다. 실험동물군 피부의 육안적인 관찰은 실험 종료후 AR-2(사삼추출물 5%)를 피부에 도포한 군에서는 100%의 발모현상이 나타났고. AR-2>AR-1>ET>PBS의 순으로 발모효과가 높았다. 실험동물군 모낭의 현미경적 소견은 AR-2를 도포한 군에서 가장 많은 수의 모낭이 관찰되었으며 또한 모낭 내의 hair shaft가 가장 많았다. 모낭의 수는 AR-2>AR-1>ET>PBS 순으로 많았다. 피부의 멜라닌 색소 침착 정도는 1일, 5일, 9일째 측정하였으며 9일째 측정한 값은 각각의 실험군에 따라 267.15(±15.03), 320.45(±75.53), 320.45(±41.94), 193.10(±22.61)이었다(p<0.05). C57BL/6는 melanocyte가 모낭에만 한정적으로 존재하고 있으며 melanin 합성이 모발성장주기와 잘 일치되어 성장기의 경우 진회색을 띠어 피부색으로 모발의 성장주기를 판정할 수 있다. 실험동물의 모발두께와 길이 변화 측정에서는 각각 실험군별로 두께(㎛)는 AR-1(사삼추출물 1%)은 31.22(±10.62), AR-2(사삼추출물 5%)는 34.82(±6.07) ET(에탄올)는 29.42(±4.94), PBS는 27.09(±5.89)(p<0.05)로 나타났다. 길이(mm)는 AR-1(사삼추출물 1%)은 4.24(±1.43), AR-2(사삼추출물 5%)는 5.11(±0.83), ET는 3.98(±0.60), PBS는 3.72(±0.40)로 유의적인 차이가 있었다(p<0.05). 모발생성 및 분포관찰은 VISIOSCAN V98을 이용하여 간접적으로 확인 하였는데 시간이 경과함에 따라 AR-2 실험군(사삼추출물5%)의 모발의 수가 늘고 분포도 조밀해짐을 확인하였다. 2. 인체피부에 적용한 피부도포임상실험 결과는 다음과 같다. 실험군의 일반사항으로 평균연령은 43.5세이며, 신장 172.3cm, 체중 70.4kg로 조사되었으며, BMI는 23.7로 보통체형으로 조사되었고, 조사대상자의 95%가 기혼자이며, 직업은 전문직 및 관리직이 80%로 가장 많았고, 월수입은 300만원 이상 65%, 교육정도는 전문대학졸이 40%로 가장 많았다. 연구대상자의 건강생활습관 정도는 평균 2.65(±1.18)로 “보통이다”인 3에 비해 다소 낮게 나타났으나 음주관련점수 3.25(±1.29)와 흡연관련점수는 3.60(±1.50)로 보통보다 좋은 것으로 조사되었다. 스트레스 정도는 전체 평균 점수가 2.88(±0.88)로 “보통이다”인 3점에 비해 비교적 스트레스 정도가 낮은 것으로 나타났으나 항목별 스트레스 점수 중 최근 3~6개월 동안의 스트레스는 3.55(±0.83)로 보통 수준보다 스트레스점수가 높게 나타났다. 연구대상자의 영양섭식관리 항목의 총 평균은 2.95(±1.08)로 “보통이다” 3점보다 다소 낮았으나 균형 잡힌 식사, 아침식사, 단 음식의 제한, 다양한 야채와 제철과일 섭취는 좋은 편으로 평가되었다. 모발상태와 관리습관에서는 탈모가 처음 발견된 시기는 30대 초반과 30대 후반이 각각 30.0%로 가장 많았으며 모발재생을 위한 방법으로는 발모제(40.0%)와 기능성 샴푸(35.0%)를 가장 선호하는 것으로 조사되었는데, 탈모의 증상이 심하지 않을수록 발모제에 대한 선호도가 큰 것으로 파악되었으나 유의적인 차이는 없었다. 실험군의 두피상태는 점수가 높을수록 두피상태가 좋지 않은 것으로 파악하였으며 총 평균이 2.78(±1.01)로 “보통이다” 3점에 비해 비교적 좋은 것으로 나타났다. 실험군의 모발상태는 3.08(±1.13)로 두피상태보다는 좋지 않은 것으로 나타났다. 두피의 상태가 나쁠수록 더 빠른 시기에 탈모가 발견된다는 유의한 결과를 나타냈다(p<0.05). 실험군의 사삼 헤어토닉 사용 전-후 모발 및 두피의 특성변화는 두피특성 중 비듬의 정도가 사전 평균이 2.60(±1.10)에서 사후 2.15(±0.99)로 감소되었고, 두피의 염증정도는 사전 2.35(±0.99)에서 사후 2.00(±0.97)으로 감소되었으나 통계적으로 유의하지 않았다(p＞0.05). 두피의 가려움 정도는 사전 평균 2.85(±0.81)에서 사후 2.20(±0.70)으로 감소되었고, 두피의 기름기도 사전 평균이 3.30(±0.92)에서 사후 2.70(±1.03)로 감소되어 사삼헤어토닉의 사용이 두피의 가려움과 기름기를 유의적으로 감소시키는 결과를 나타났다(p<0.05). 사삼헤어 토닉 사용 후 모발상태 특성은 모발의 두께가 사전 3.50(±0.95)에서 사후 3.05(±1.05)으로, 백발의 양 변화는 사전 2.80(±1.11)에서 사후 2.40(±0.94)으로, 머리숱의 정도 변화는 사전 3.75(±0.97)에서 사후 3.55(±0.83)로, 곱슬거림 정도는 사전 2.85(±1.27)에서 사후 2.50(±1.36)으로 모든 항목에서 평균적으로 감소되었으나 통계적으로 유의한 차이는 없었다. 결론적으로 사삼헤어토닉의 사용은 두피의 가려움과 기름기를 줄이는데 유의한 효과가 있는 것으로 나타났으며, 통계적으로 유의미하지는 않았으나 전반적으로 두피와 모발에 좋은 영향을 주는 것으로 조사되었다. 사삼헤어토닉 품평서 평가는 사용의 용의성은 3.90(±0.64), 두피자극성 3.40(±0.99), 청량감 4.00(±0.65), 세정성 3.15(±0.49), 향 3.75(±0.55), 구입의사는 3.65(±0.59)로 대체로 좋은 평가를 받은 것으로 나타났다.|This research has been conducted to explore the effect of Adenophora Radix, as a natural substance, on hair growth and to provide basic data on the development potential of Adenophora Radix as a cosmetic product for preventing hair loss. Accordingly a hair growth efficiency test and a clinical test on application to human skin were carried out through a mouse model. In relation to the hair growth efficiency test which was carried out through the mouse model, 6 weeks old C57BL/6 male mice were used as the test animal and a powder obtained from enrichment and lyophilization of a filtrate of adenophora triphylla var. japonica soaked in 70% ethanol liquid was used for the Adenophora Radix sample. As test substances, AR-1(1% Adenophora Radix extract), AR-2(5% Adenophora Radix extract), ethanol and PBS were used. Observations were carried out on weight variation, skin examination with naked eyes, fractography of hair follicle, degree of melanin pigmentation, change in hair thickness and length and regeneration and distribution of hair by applying each sample to the hair loss area of the test animal. The clinical test on application to human skin was carried out on males in their 20s to 40s suffering from hair loss. Adenophora Radix hair tonic containing AR-2(5% Adenophora Radix extract), which had a statistically significant hair growth effect compared to the control group in the animal test, was manufactured and applied to the hair loss area for 5 weeks. As a research method, respondents were asked to fill in a survey and an Adenophora Radix hair tonic product quality assessment form by themselves before and after the test and the changes in hair and scalp before and after using the Adenophora Radix hair tonic was analyzed using the collected surveys and forms. The survey was composed of questions on general features, life style and health status, extent of stress, nutrition and diet management, hair condition and scalp management habit and current scalp and hair condition. The Adenophora Radix hair tonic product quality assessment form was composed of 6 type of questions, namely ease of use, skin irritation, refreshing feeling, cleaning ability, scent preference and opinion on purchasing the product and respondents were asked to answer these questions. 1. The hair growth efficiency test result obtained through a mouse model is as follows. Mice used in the test showed the following change in weight (g): in R-1(1% Adenophora Radix extract) 23.52(±0.48), in AR-2(5% Adenophora Radix extract) 24.12(±0.74), in ET(ethanol) 24.00(±0.45), and in PBS 23.52(±0.54). There was no difference in the weight of test mice between different test materials, namely the 4 types of Adenophora Radix extract liquid and there was also no statistically significant difference in amount of weight increase. On skin observation with naked eyes, after the test was completed, a 100% hair growth was shown in a group which AR-2(5% Adenophora Radix extract) was applied and a higher hair growth effect was shown in the following order AR-2>AR-1>ET>PBS. As a result of microscopic examination on test animal groups, the highest number of follicles and hair shaft in the follicles were shown in the group which AR-2 was applied. Higher number of follicles were shown in the following order AR-2>AR-1>ET>PBS. In relation to the degree of melanin pigmentation of skin, an observation was carried out on the 1st, 5th and 9th day and the value obtained on the 9th day was 267.15(±15.03), 320.45(±75.53), 320.45(±41.94), 193.10(±22.61) respectively according to each test group(p<0.05). C57BL/6 shows that melanocyte only exists restrictively in follicles and allows to determine the growth period of hair by skin colour since melanin synthesis confirms well with the hair growth period and shows a dark grey color during the growth period. As a result of measuring the change in hair thickness and length, in AR-1(1% Adenophora Radix extract) 31.22(±10.62), in AR-2(5% Adenophora Radix extract) 34.82(±6.07) in ET(ethanol) 29.42(±4.94) and in PBS 27.09(±5.89)(p<0.05) thickness(㎛) were shown according to each test group and a statistically significant difference was shown in length(mm), namely in AR-1(1% Adenophora Radix extract) 4.24(±1.43), in AR-2(5% Adenophora Radix extract) 5.11(±0.83), in ET 3.98(±0.60), and in PBS 3.72(±0.40) (p<0.05). The observation on hair regeneration and distribution was carried out indirectly using a VISIOSCAN V98 and was found that as time passed, the amount of hair increased and became more densely distributed in the AR-2 test group (5% Adenophora Radix extract). 2. The clinical test result on application to human skin is as follows. General features of the test group were that they had an average age of 43.5%, height of 172.3cm and weight of 70.4kg. In addition, BMI was observed to be 23.7 namely average built and 95% of the respondents were married. Respondents were mostly involved in professional or managerial occupation amounting to 80% of the respondents and 65% of the respondents had a monthly income of 3 million Korean Won or more. For educational background, college graduates were most common being 40%. In relation to the degree of healthy life habit, respondents were shown to be slightly below 3 "normal" the average being 2.65(±1.18) but the grades related to drinking and smoking were above normal being 3.25(±1.29) and 3.60(±1.50) respectively. The extent of stress was shown to be relatively less than 3 "normal" having the overall average of 2.88(±0.88) but amongst the stress items, the extent of stress within the recent 3~6 months were shown to be higher than the normal level being 3.55(±0.83). The total average of nutrition and diet management items was slightly below 3 "normal" being 2.95(±1.08) but balanced diet, breakfast, food limitation and consumption of various vegetables and seasonal fruits were all evaluated to be good. In relation to hair condition and management habit, most respondents had their first hair loss either in early 30s or late 30s both of them amounting to 30% of the respondents. Respondents preferred using hair growth products (40.0%) and functional shampoos (35.0%) most as a method of regenerating hair. It was also shown that people with less serious hair loss tend to prefer hair growth products more but no statistically significant difference was shown. On the scalp condition of the test group, it was showed that respondents with higher grades had bad scalp condition but was relatively better than 3 "normal" having the general average of 2.78(±1.01). The hair condition of the test group was shown to be 3.08(±1.13) worse than the scalp condition. A statistically significant result that, the worse the scalp condition, the earlier the hair loss is found, was observed (p<0.05). In relation to the change in features of hair and scalp before and after using the Adenophora Radix hair tonic, it was found that amongst scalp features, the degree of dandruff was reduced from 2.60(±1.10) (average before use) to 2.15(±0.99) (average after use), the degree of inflammation was reduced from 2.35(±0.99) (average before use) to 2.00(±0.97) (average after use) but did not amount to a statistically significant difference(p＞0.05). The degree of itching of the scalp was reduced from 2.85(±0.81) (average before use) to 2.20(±0.70) (average after use) and the greasiness of scalp was also reduced from an average before use of 3.30(±0.92) to an average after use of 2.70(±1.03) proving that Adenophora Radix hair tonic has a statistically significant result of reducing scalp itching and greasiness (p<0.05). In relation to the features of hair condition, after using the Adenophora Radix hair tonic, hair thickness increased from 3.50(±0.95) (before use) to 3.05(±1.05) (after use), amount of white hair changed from 2.80(±1.11) (before use) to 2.40(±0.94) (after use), the amount of hair changed from 3.75(±0.97) (before use) to 3.55(±0.83) (after use), frizziness changed from 2.85(±1.27) (before use) to 2.50(±1.36) (after use), showing that although a reduction was shown in all items in average, there was no statistically significant difference. In conclusion, it was found that use of Adenophora Radix hair tonic has a statistically significant effect in reducing itchiness and greasiness and has a good effect in general to hair and scalp although did not amount to a statistically significant effect. The Adenophora Radix hair tonic product quality assessment form received a good feedback in general, having the following results: 3.90(±0.64) for ease of use, 3.40(±0.99) for skin irritation, 4.00(±0.65) for refreshing feeling, 3.15(±0.49) for cleaning ability, 3.75(±0.55) for scent preference and 3.65(±0.59) for opinion on purchasing the product.