• 대한의생명과학회지
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• 제16권4호
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• pp.349-357
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• 2010

We have examined the histological changes of skin during hair follicle growth after depilation in C57BL/6N mice. We first studied on histological changes of number of mast cells and thickness of skin during hair follicle growth periods (telogen, 1 day, 3 day, 5 day, 10 day, 14 day, 17 day and 21 day after depilation) by toluidine blue, Giemsa and H&E staining methods. We second studied immunoreactive density of cytokines and Brdu labeled cells in skin during hair follicle growth periods after depilation in C57BL/6N mice by immunohistochemical methods. The histological changes on skin thickness was increased from telogen to 14 day. The number of mast cells was decreased in 3,5 and 10 day and increased in 14, 17 and 20 day after depilation. Immunoreactive density of cytokines [protein kinase C-${\alpha}$ (PKC-${\alpha}$), c-kit, and vascular endothelial growth factor (VEGF)] in 1, 3, 5, 10, and 14 day after depilation was mildly stained in bulge and cutaneous trunci m., but immunoreactive density of cytokines in 17 and 21 day was heavily stained in epidermis, bulge, outer root sheath (ORS), inner root sheath (IRS) and cutaneous trunci m.. Immunoreactive density of Brdu labeled cells in skin in 1 and 3 day was heavily stained in bulge, epidermis and connective tissue under the cutaneous trunci m.. In all periods, immunoreactive density of Brdu labeled cells in skin was heavily stained in bulge, subcutaneous tissue, cutaneous trunci m, ORS and IRS. These experiments suggest that histological changes related to hair follicle growth elevated mast cell counts, skin thickness and epidermis thickness and heavily stained immunoreactive density of cytokines and Brdu labeled cutaneous trunci m. and connective tissue under the cutaneous trunci m. after depilation in C57BL/6N mice.

• 생약학회지
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• 제52권3호
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• pp.134-142
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• 2021

This study was conducted to evaluate the effects of Dictyota coriacea extract and its active components such as 1,9-dihydroxycrenulide and epiloliolide on the hair growth. Treatment with D. coriacea extract and the hexane and EtOAc fractions of D. coriacea extract significantly increased the proliferation of dermal papilla cells (DPCs), a central regulator of the hair cycle. Especially, 1,9-dihydroxycrenulide and epiloliolide from D. coriacea extract, caused an increase in the DPC proliferation. When isolated rat vibrissa follicles were treated with 1,9-dihydroxycrenulide or epiloliolide for 21 d, the hair-fiber lengths for the vibrissa follicles increased. When examined the activity of 5α-reductase, which converts testosterone to dihydrotestosterone (DHT), a main cause of androgenetic alopecia, the several solvent fractions of D. coriacea extract significantly decreased the 5α-reductase activity while 1,9-dihydroxycrenulide and epiloliolide scarcely inhibited 5α-reductase activity. In addition, we found that the D. coriacea extract and several solvent fractions of D. coriacea extract could not act as a K_ATP channel opener, which could be a contributory factor in the effect on hair growth. These results suggest that D. coriacea extract and 1,9-dihydroxycrenulide and epiloliolide, principals of D. coriacea, have the potential to treat alopecia via the proliferation of DPCs.

• 동의생리병리학회지
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• 제23권5호
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• pp.1116-1124
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• 2009

To determine whether topical application of trimix extracts of Mylabris phalerata Pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata lead to affects on the hair growth activity in C57BL/6N mice. To examine the hair growth activity of the extracts of Mylabris phalerata Pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata gross, microscopic, and immunohistochemical method were performed. In order to examine the mRNA expression of hair growth related substance, RT-PCR method was performed. Experimental group I on day 14, The most extensive hair growth activity was observed in whole skin area of all the mice whose hair had been clipped. Brdu immunoreactive cells of all the experimental groups were more heavily stained in epidermis, bulge, outer root sheath, inner root sheath, subcutaneous tissue, hair bulb and cutaneous trunci muscles than that of control group on day 12 of hair growing cycle in C57BL/6N mice. VEGF immunoreactive density of all the experimental groups was more heavily stained in epidermis, bulge and cutaneous trunci muscles than that of control group on day 12. FGF and c-kit immunoreactive cells of all the experimental groups were heavily stained in epidermis, outer root sheath, inner root sheath and cutaneous trunci muscles on day 12. PKC-$\alpha$ immunoreactive density of all the experimental groups was mildly stained in epidermis and cutaneous trunci muscles than that of control group on day 12. On day 12, the expression of bFGF (138%, 119%, 120%), VEGF (146%, 144%, 133%), IGF-1 (165%, 141%, 119%) and PLI (121%, 116%, 123&) in each experimental groups was more increased than that of control group. On day 16, The expression of IGF-1 (126%, 149%, 151%) in all the experimental group was more increased than that of control group (100%). The expression of bFGF (92%, 94%) and VEGF (101%, 97%), PL1 (102%, 109%) in all the experimental group was more decreased than that of experimental group I, II on day 12. But the expression of bFGF (109%) and VEGF (127%), and PL1 (105%) in each experimental group III was more increased than that of control group (100%). These experiments suggest that trimix extracts of Mylabris phalerata pall., Arisaematis Rhizoma and Pinelliae Rhizoma Ternata may stimulate the topical hair growth activity and its experimental group I can be useful for treatment of alopecia areata.

• 대한한의학회지
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• 제31권1호
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• pp.138-152
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• 2010

Objective: Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) is one of the representative prescriptions for invigoration of vitality and nourishing of the blood. This study was carried out to investigate the effects of Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) on hair regrowth and cytokine changes in a shaving model of C57BL6 mice. Method: Five-week-old mice were acclimated for 1 week at a temperature between $21-23^{\circ}C{\acute{E}}$, 40-60% relative humidity, and 12h of a light/dark cycle before beginning of the experiment. There were two groups including normal saline (control) and a positive control of oral intake of Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) extract (sample) in 18 female mice. The test compounds were topically treated once a day over 14 days. The hair regrowth was photographically and histologically determined during the experimental period of 14 days. Revelation of TGF-${\beta}1$ and EGF were also determined using immunohistochemistry. In addition to that, IFN-$\gamma$, IL-4 and IL-10 were determined in serum. Results: Hair regrowth in the sample group was promoted earlier and faster than the control group, as shown by concentrations of hairs and thick-hair ratio in the sample group. TGF-${\beta}1$ was not revealed in either control or sample group. EGF was strongly positive in out root sheath of some thick hair of the sample group. Serum IFN-$\gamma$ was significantly decreased in the sample group compared with the control group at 7 experimental days. However, it was not significantly decreased at 14 experimental days. Serum IL-4 was significantly increased in the sample group compared with the control group at 7 experimental days. However, it was not significantly decreased at 14 experimental days. Serum IL-10 was decreased in the sample group compared with the control group, but with no real statistical significance. Conclusion: These results suggest that Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) has hair growth promoting activity and it can be used for treatment of alopecia. Also, these effects relate to EGF revelation of hair roots, a decrease in serum IFN-$\gamma$, and an increase of serum IL-4.

• 대한화장품학회지
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• 제43권4호
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• pp.281-287
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• 2017

피부에 가해지는 스트레스는 헤어조절 및 사이클에 직 간접적으로 중요한 영향을 미친다고 알려져 있다. 특히, 모근세포는 스트레스에 의한 부신피질관련호르몬과 세포손상 및 사멸과 밀접한 관련이 있다고 보고되고 있지만, 현재까지 실험적으로 입증된 사실은 매우 제한되어 있다. 보고에 의하면, 부신피질자극호르몬방출인자가 증가되면 모근세포의 마이토콘드리아 활성을 저해하여 초기단계의 세포사멸을 가져올 수 있다고 임상학적으로 보고된바가 있다. 특히 아토피 피부염으로 인한 스트레스는 부신피질자극호르몬방출인자와 부신피질관련 호르몬의 양을 증가시키며, 이는 모발의 outer epithelial sheath에 영향을 준다고 알려져 있으며, 이러한 스트레스의 변화는 마이토콘드리아 손상을 초래하여 초기단계세포손상을 준다고 한다. 따라서 본 연구는 아토피피부염스트레스가 피부의 모근세포에 주는 영향에 대하여 연구를 하였는데, 이에 대한 연구는 현재까지 전무한 실정이다. 우리는 NC/Nga 마우스에 2,4-dinitrochlorobenzene (DNCB)로 아토피피부염을 유발 후, 피부 스트레스 생성에 의한 초기단계 세포손상을 스트레스관련 인자, 부신피질자극호르몬방출인자 및 그 관련 인자, annexin V 및 마이토콘드리아 반응을 이용하여 연구하였다. 그 결과, 아토피피부염에 의한 스트레스는 체내의 부신피질 자극호르몬방출인자 및 관련인자의 활성을 증가시킬 뿐 아니라, 모근세포에 영향을 주어 초기단계세포사멸을 초래하는 것으로 나타났다. 이는 아토피피부염관련 헤어손상을 일으킨다는 중요한 연구결과를 보고하는 바이며, 부신피질자극호르몬 조절관련 의약품 및 화장품 등과 같은 보조적 요법이 필요함을 제안한다.

• 생명과학회지
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• 제26권11호
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• pp.1345-1354
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• 2016

원형탈모증은 흔하게 발병하고 두피와 전신에 모발의 탈락을 일으키는 모낭조직 특이 자가면역질환이다. 모낭은 자체적으로 면역체계와 내분비 환경을 가지고 각 모발주기 단계에 따라 다른 면역 상태를 나타내는 특이한 기관이다. 성장기 모낭의 면역특권의 파괴는 모낭상피 MHC class I 발현과 자가반응성 CD8+T세포에 대한 자가항원 발현을 유도하는 자가면역의 공격을 일으키고 원형탈모증을 유발한다. 임상적 실험적 연구에 의하면, 심리적 스트레스도 모낭 면역/호르몬 체계에 영향을 미쳐 원형탈모증의 유도에 관여할 수 있다고 지적한다. 원형탈모증의 핵심적인 병리기전은 면역특권 수호자(ACTH, ${\alpha}-MSH$와 $TGF-{\beta}$ 등), 자연살해세포그룹 2D-양성(NKG2D+) 세포(NK 세포와 CD8+T 세포 등)와 스트레스 호르몬(CRH와 substance P)과 관련되어 있다. 효과적인 치료법은 여전히 요구되고 있다. 앞으로 치료목표 중의 하나는 스트레스를 포함한 모낭 면역특권을 개선하는 것일 것이다. 최근 연구는 건선, 아토피피부염, 류마티스 관절염 같은 다른 자가면역질환에서 사용되는 JAK억제제와 면역조절제, Tregs, 혈소판풍부혈장요법, 스타틴과 프로스타글란딘 유사제가 원형탈모증에 효과적이라고 보고하였다. 본 논문은 모낭주위 내분비/면역과 관련된 발병기전에 대한 새로운 이해와 원형탈모증의 새로운 치료법에 대해 고찰하였다.

• Biomolecules & Therapeutics
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• 제29권2호
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• pp.211-219
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• 2021

Alopecia is a distressing condition caused by the dysregulation of anagen, catagen, and telogen in the hair cycle. Dermal papilla cells (DPCs) regulate the hair cycle and play important roles in hair growth and regeneration. Myristoleic acid (MA) increases Wnt reporter activity in DPCs. However, the action mechanisms of MA on the stimulation of anagen signaling in DPCs is not known. In this study, we evaluated the effects of MA on anagen-activating signaling pathways in DPCs. MA significantly increased DPC proliferation and stimulated the G2/M phase, accompanied by increasing cyclin A, Cdc2, and cyclin B1. To elucidate the mechanism by which MA promotes DPC proliferation, we evaluated the effect of MA on autophagy and intracellular pathways. MA induced autophagosome formation by decreasing the levels of the phospho-mammalian target of rapamycin (phospho-mTOR) and increasing autophagy-related 7 (Atg7) and microtubule-associated protein 1A/1B-light chain 3II (LC3II). MA also increased the phosphorylation levels of Wnt/β-catenin proteins, such as GSK3β (Ser9) and β-catenin (Ser552 and Ser675). Treatment with XAV939, an inhibitor of the Wnt/β-catenin pathway, attenuated the MA-induced increase in β-catenin nuclear translocation. Moreover, XAV939 reduced MA-induced effects on cell cycle progression, autophagy, and DPC proliferation. On the other hand, MA increased the levels of phospho (Thr202/Tyr204)-extracellular signal regulated kinases (ERK). MA-induced ERK phosphorylation led to changes in the expression levels of Cdc2, Atg7 and LC3II, as well as DPC proliferation. Our results suggest that MA promotes anagen signaling via autophagy and cell cycle progression by activating the Wnt/β-catenin and ERK pathways in DPCs.

• 대한한의학회지
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• 제43권1호
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• pp.18-32
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• 2022

Objectives: This study aimed to examine the safety, effects on proliferation of hair papilla cells, and anti-inflammatory and antioxidant mechanisms of Artemisia sieversiana Ehrh. ex Willd. (AS) extract. Methods: Safety tests through purity testing, acute toxicity tests, and repeated toxicity tests were performed using AS extract (ASE) which had been dried for over two years. Cell culture and proliferation tests were conducted; VEGF (vascular endothelial growth factor), bFGF (basic fibroblast growth factor), and EGF (epidermal growth factor) and protein expression analyses were performed for mechanistic evaluation; and inhibitory effects of ASE on the RNA expression of testosterone, 5𝛼-reductase, and aromatase was assessed. The anti-inflammatory and antioxidant efficacy of ASE was confirmed by measuring the levels of nitric oxide, inflammatory mediators (TNF-𝛼 and PGE2), inflammatory cytokines (IL-1𝛽, IL-6, and IL-8), and chemokine MCP-1. Results: The safety of ASE was confirmed. The mechanism of cell proliferation in human hair follicle dermal papilla cells involved the promotion of VEGF, bFGF, and EGF expression. ASE decreased mRNA expression of testosterone, 5𝛼-reductase, and aromatase-1 in a concentration-dependent manner. PGE2 and TNF-𝛼 production by inflammatory mediators was also significantly decreased in a concentration-dependent manner, and inflammatory cytokine and chemokine expression was inhibited. Conclusions: ASE is suggested to promote papillary cell growth at the cellular level, to suppress expression of various enzymes involved in hair cycle and cell death, and to inhibit hair loss through anti-androgen, anti-inflammatory, and antioxidant effects.

• 한국식품과학회지
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• 제54권1호
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• pp.43-51
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• 2022

본 연구에서는 자작나무 증포 추출물의 탈모 조절 활성 분석을 위해 in vitro (인간모유두세포) 및 in vivo (C57BL/6N 마우스) 모델을 이용하여 모발의 성장 효과를 평가하였다. 찌고 말리는 과정을 반복하는 증포 차수 별 함유 성분의 변화가 관찰되어 새로운 추출법의 가능성을 확인하였다, 즉, 1회-5회 증포 후 관찰된 성분의 변화는 3회 증포 추출물(BPE3)에서 안정적인 추출 수율, 높은 페놀화합물 함량 및 항산화 활성을 가지는 것으로 확인되었다. 또한, 발모 주기의 전 과정에 관여하는 모유두세포에 BPE3를 처리하였을 때 유의한 수준의 FGF7과 Wnt7b 발현을 증가시켜 모발 성장 촉진과 모발의 성장기 개시를 도울 것으로 판단되었다. In vivo 마우스 모델에 12일 간 BPE를 도포하여 관찰한 결과 6일 경과 시 양성대조군(MXD 및 PTN)과 유사한 수준으로 단모의 성장이 관찰되었으며, 9일 경과 시 높은 밀도의 발모가 진행되기 시작하여 12일 경과 시 미처리 대조군에 비해 BPE3군에서 고른 발모가 관찰되었다. H&E 염색을 통한 각 군별 피부조직의 변화는 BPE3군에서 뚜렷이 나타났으며, 특징적으로 단위면적 당 많은 모낭(hair follicle)의 형성과 모간부(hair shaft)의 신장이 관찰되어 안정적으로 모발의 성장기로 진입한 것으로 판단되었다. 피부조직의 유전자발현 추가 분석 시 FGF7, VEGF, 및 Wnt7b 유전자가 유의하게 증가하여 모발성장, 분화, 모낭줄기세포 활성을 유도하여 모발성장을 촉진시킨 것으로 생각된다. 또한, BPE3가 LPS로 유도된 RAW264.7 세포의 염증인자(iNOS, IL-6 및 COX2) 발현을 저해하여 자가면역 등 염증성 탈모억제에 긍정적 역할을 할 것으로 판단된다. GC-MS 분석을 통해 확인한 betulin과 불포화지방산 등 저분자 물질은 BPE3가 나타낸 약리활성을 방증하였다. 결론적으로, 자작나무 3회 증포 추출물인 BPE3는 모유두세포의 발모 주기를 촉진할 뿐 아니라 두피의 염증 환경에서 휴지기를 단축시켜 정상적 발모를 돕는 소재로서 높은 잠재력을 나타냈다.

• 한방재활의학과학회지
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• 제19권4호
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• pp.95-113
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• 2009

Objectives : Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) is a hair care product which is composed of ten plant extracts used in oriental medicine. This study was carried out to investigate the effects of Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) on hair regrowth and cytokine changes in a shaving model of C57BL/6 mice. Materials and Methods : Five-week-old mice were acclimated for 1 week at a temperature between $21-23^{\circ}C$, 40-60% relative humidity, and 12h of a light/dark cycle before beginning of the experiment. There were three experimental groups including 50% ethanol (EtOH, control), a positive control of 3% Minoxidil, and 30% Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) in 50% ethanol in 18 female mice. The test compounds were topically treated once a day over 12 days. The hair regrowth was photographically and histologically determined during the experimental period of 12 days. Revelation of EGF, $TGF-{\beta}1$ and IL-6 in hair follicle were also determined using immunohistochemistry. In addition to that, IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ in skin tissue were determined using enzyme-linked immunosorbent assay(ELISA). Results : Hair regrowth in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups was promoted earlier and faster than the control group. Concentrations of hairs and thick-hair ratio in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups were promoted than the control group. EGF was moderately positive in hair follicle of 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups, but negative in the control group. $TGF-{\beta}1$ was not significantly difference between the groups. IL-6 in hair follicle of Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) group was negative, but weakly positive in 3% Minoxidil and control group. IL-6 and $IL-1{\beta}$ in skin tissue were significantly decreased in Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) group, but there was not significantly decreased in 3% Minoxidil and control group. $TNF-{\alpha}$ in skin tissue was significantly decreased in 3% Minoxidil and Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) groups. Conclusions : These results suggest that Saengbal-eum-II($Sh{\bar{e}}ngf{\grave{a}}-y{\breve{i}}n-ll$) has hair growth promoting activity and it can be used for treatment of alopecia. And these effects relate to EGF revelation of hair follicle and a decrease IL-6, $TNF-{\alpha}$, and $IL-1{\beta}$ in skin tissue.