| Alopecia or hair loss is one of the common diseases in dermatology,although it is not a life threatening illness but it can bring great painful suffering on human mental stress over life time.Up to date,finasteride and minoxidil are two major medicines to treat alopecia.They show therapeutic effects but also have significant adverse effects.In this study,we selected Anemarrhena asphodeloides and Phaeodactylum tricornutum with 5?-reductase inhibition and keratinocytes renewal promoting effects.Single cell cultures,hair follicle tissue culture in vitro and C57BL/6 male mice in vivo were applied to evaluate the activities of the two extracts for hair follicle cycle signals,hair follicle structure strength markers,as well as melanin production and transportation related protein factors,and to further reveal the molecular mechanisms of hair follicle promoting effects of PTE and AAE.In addition,hair and scalp health conditions were evaluated with a scalp care formula containing PTE and AAE by a comprehensive physiological and biochemical clinical testing procedures.The main results of this project are as the followings:(1)Active components extract and analysis from Phaeotrichum tricornutum and Anemarrhena asphodeloidesPhaeodactylum tricornutum was extracted with ethanol in nitrogen atmosphere.The extraction rate of Phaeodactylum tricornutum was 0.985% containing octadecane unsaturated fatty acids as the main components.Anemarrhena asphodeloides was extracted with water/water ethanol solution.The yield of total polysaccharide is 22.642%;total saponin 10.564%,in which contained 28.6% timosaponin BII and 2.75% mangiferin;total flavone were 6.378%,in which contained 21.60% mangiferin and 3.46% isomangiferin.(2)PTE and AAE effects on hair follicle growth based on related cell culture systemshuman hair follicle related cells system were used for in vitro testing including human hair papilla cells(DPCs),human hair germinal matrix cells(HFGMCs),human hair follicle outer root sheath cells(HFORSCs),human hair follicle inner root sheath cells(HFIRSCs)and human epidermal melanocytes(HEM).PTE and AAE had certain growth promoting effects on hair follicle unit related cells,while some components had inhibitory effects at high concentrations.The total timosaponins,timosaponin BII and mangiferin of AAE at 100?g/ml caused decreases of cell density and appeared atrophy of hair follicle in vitro.The increase in regrowing area of hair and hair follicles numbers and the improvement of hair follicle morphology in the group applied with 0.5% timosaponin BII indicated an induction of the anagen phase in telogenic mice skin,which were comparative to the 2% minoxidil treatment.The immunohistochemical analysis detected an increase in the expression of ?-catenin and Wnt10 b,supporting the theory of the activation of ?-catenin/Wnt pathway was one of the pathways that related to anagen phase induction.(3)PTE and AAE effects on signal transductions and major proteins and factors of hair strength on hair follicle related cellsPTE is able to promote the expression of several keratins in HFORSCs at various degrees,especially at 0.25?l/ml.PTE at 0.125-0.5?l/ml was able to improve the activity of HEM significantly,0.5?l/ml PTE increased ERK by 56%.Timosaponin BII and polysaccharides of AAE showed inhibitory effects on IGF-1 and cyclin D1 signals in DPCs,which may lead to the inhibition of hair follicle growth cycle.The effects of AAE on HFGMCs can promote or inhibit the growth and development of hair follicles through Wnt-10b/?-Catenin and phospo-GSK-3? signals.AAE was able to inhibit hair follicle growth through cellular signals of ?-catenin,Akt and Wnt-10 b on HFGMCs,while AAE showed d uncertain effects on HFIRSCs through ?-catenin,Akt and phospo-GSK-3? signals.The reason may due to the higher doses of AAE applied to cultures that brought more negative effects on the cell growth.(4)A scalp formula contains PTE and AAE efficacy on human hair growth30 human subjects were recruited and applied a scalp care formula containing PTE and AAE for 28 days.The overall number of hair falls,the scalp oil level,the scalp erythema,the scalp discomfort feeling,and the dandruff level were all reduced significantly after 7 days post the formula applications,while the hair appearance showed improvement significantly.The overall trends of hair health improvements throughout the clinical period but showed no statistical significance,of which may due to the limited duration of the study length,and/or less enough study subjects,as well as the nature of the individual physiological diversity and variations.The hair follicle hydrocortisone level increased significantly in the first 14 days,and decreased significantly from the fourth week of the application.This result suggested that the test formula may promote hair follicle growth through the increase of hydrocortisone level,while the decrease may suggest help to reduce the metabolism of dandruff formation.In conclusion,proper concentrations of the PTE and AAE were able to promote the hair follicle growth and hair loss prevertion.The results provide further helps to understand the mechanisms of hair growth in vitro. |
PDF Full Text Request